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  • Saccharomyces cerevisiae  (112)
  • Coleoptera
  • Springer  (190)
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  • 1995-1999  (91)
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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Entomologia experimentalis et applicata 80 (1996), S. 453-460 
    ISSN: 1570-7458
    Keywords: Coleoptera ; Curculionidae ; Metamasius hemipterus ; aggregation pheromone ; redundancy ; field trapping ; sex-ratio
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The five synthetic pheromone components of the West Indian sugarcane borer (WISB), Metamasius hemipterus (L.) (Coleoptera, Curculionidae) were tested in the field. The combination of sugarcane (SC) and the major pheromone compound, 4-methyl-5-nonanol (1) was attractive. However, the addition of 2-methyl-4-heptanol (2) or 2-methyl-4-octanol (3) was required to reach high catch levels while 5-nonanol (4) or 3-hydroxy-4-methyl-5-nonanone (5) did not enhance WISB attraction. The redundancy phenomenon, here reported for the first time in rhynchophorinous species, was observed between compounds 2 and 3. SC +1 +3 was more attractive than living male baits, however, the sex-ratio of the catches was equivalent between both treatments. The sex-ratio of catches was affected by the qualitative composition of the pheromone formulation. Compound 3 had a sexual role, attracting more females while 5 seemed to play an aggregation role, luring both sexes in the same proportion.
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  • 2
    ISSN: 1570-7458
    Keywords: Oryzaephilus surinamensis ; Coleoptera ; Cucijidae ; population density ; semiochemicals ; lethal secretions
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
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  • 3
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    Entomologia experimentalis et applicata 69 (1993), S. 13-20 
    ISSN: 1570-7458
    Keywords: Artificial diets ; Coccinellidae ; rearing ; biological control ; Chilocorus spp. ; Coleoptera
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Promising diets were screened and the most successful modified with additives used in artificial diets for other entomophagous insects. Two suitable diets were obtained, one for adults and one for larvae ofChilocorus nigritus (Fabricius) (Coleoptera: Coccinellidae). They were still inferior to natural prey and not adequate as the sole food source for rearing consecutive generations. They are valuable as substitute food in the insectary during shortages of natural prey. Oleander scaleAspidiotus nerii Bouché andAsterolecanium miliaris (Boisduval) were evaluated as natural prey forC. nigritus and two other potential biocontrol agents in southern Africa,C. bipustulatus (Linnaeus) andC. infernalis Mulsant.A. nerii andA. miliaris were suitable for all life stages ofC. nigritus and adults ofC. bipustulatus andC. infernalis. A. miliaris was inadequate for larvae ofC. bipustulatus andC. infernalis.
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  • 4
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    Entomologia experimentalis et applicata 79 (1996), S. 355-362 
    ISSN: 1570-7458
    Keywords: aerial pollution ; heavy metals ; sulphur dioxide ; feeding preference ; Salix ; Melasoma lapponica ; Coleoptera ; Chrysomelidae ; population density ; induced resistance ; plant communication ; Kola Peninsula
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We studied preference for willows along a pollution gradient on the Kola Peninsula, Russia, by the leaf beetle, Melasoma lapponica. Multiple tests with leaf disks demonstrated low preference for Salix borealis, S. caprea and S. phylicifolia from the plot situated 14 km from the smelter, in comparison with conspecific plants from plots situated at 1 and 29 km distances. This pattern was observed when testing beetles orginating from any plot both in 1993 and 1994, using both young and mature leaves of S. borealis. Although fumigation of S. borealis with realistic SO2 concentration (100 μg/m3) increased plant palatability, preference for plants from our study plots did not correlate with plot-specific mean SO2 concentrations. Furthermore, no correlation with foliar concentrations of the main metal pollutants (Ni and Cu) was found. Palatability of plants was negatively correlated with population density of M. lapponica, which peaked in the moderately polluted plot 14 km from the smelter. Within this plot, beetles clearly preferred non-damaged bushes of S. borealis to previously damaged bushes. We therefore conclude that low preference of S. borealis from the moderately polluted area was caused by plant resistance induced by severe damage from M. lapponica in previous years rather than by pollution impact. However, S. caprea and S. phylicifolia had little damage from M. lapponica, and low palatability of these species in the moderately polluted plot suggests changes in plant quality similar to changes in heavily damaged bushes of S. borealis.
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  • 5
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    Entomologia experimentalis et applicata 80 (1996), S. 160-162 
    ISSN: 1570-7458
    Keywords: Phyllotreta nemorum ; Barbarea vulgaris ; Coleoptera ; Chrysomelidae ; Cruciferae ; resistance ; genetics of insects ; inheritance ; Y-linkage
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
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  • 6
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    Entomologia experimentalis et applicata 66 (1993), S. 161-169 
    ISSN: 1570-7458
    Keywords: avoidance behaviour ; capture efficiency ; Coleoptera ; time-lapse video ; visual recording
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Pitfall trap capture forSitophilus oryzae (L.),S. zeamais (Motschulsky),S. granarius (L.),Tribolium confusum (Duval),T. castaneum (Herbst);Oryzaephilus surinamensis (L.),Cryptolestes pusillus (Schonherr),Rhyzopertha dominica (Fabr.), andProstephanus truncatus (Horn) in millet was assessed by visual and time-lapse video recordings. The behaviour of different beetle species in arenas containing millet was monitored over 24 h and the frequency of encounters with the trap rim resulting in capture recorded. The capture efficiency of four types of pitfall traps (i.e. polystyrene, polythene, glass and tin-plated steel can) with rims exposed or submerged below the millet surface level were compared. Capture was related to beetle size, locomotory rate, and beetle behaviour at the trap rim as well as trap design and placement. The lighter and smaller species were least captured. Glass jars were more effective than plastic and metal containers. Traps placed with their rims submerged below the grain surface level were more efficient than those with rims exposed. Capture rate was unrelated to trap size. The frequency of encounters with trap rims was not correlated with capture rate. Three types of avoidance behaviours at the trap rims i.e. probing, skirting and spontaneous retreat, were related to capture rate, spontaneous retreat being the most effective escape mechanism and probing least.
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  • 7
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    Entomologia experimentalis et applicata 66 (1993), S. 191-196 
    ISSN: 1570-7458
    Keywords: Leptinotarsa decemlineata (Say) ; Coleoptera ; Chrysomelidae ; distribution ; diapause ; overwintering ; mortality
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
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  • 8
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    Entomologia experimentalis et applicata 80 (1996), S. 231-237 
    ISSN: 1570-7458
    Keywords: Leptosphaeria maculans ; Peronospora parasitica ; fungal pathogens ; Phyllotreta cruciferae ; flea beetles ; Coleoptera ; Chrysomelidae ; Cruciferae ; chitinase ; myrosinase ; allocation costs ; plant defense mechanisms
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We used artificial selection experiments to study genetic allocation costs and physiological mechanisms of resistance to herbivory and fungal disease. Genetic costs to resistance were present in some instances and absent in others. Genetic resistance to the fungal pathogen, Leptosphaeria maculans was cost-free, while resistance to Peronospora parasitica showed a negative genetic correlation between disease resistance and growth rate. Leptosphaeria resistant genotypes had 13% higher chitinase activity. Genetic increases in myrosinase activity were correlated with increased resistance to flea beetles (Phyllotreta cruciferae), but resulted in lower plant fecundity, presumably due to production costs of myrosinase. Genetic costs of resistance may maintain genetic variation in natural plant populations. These studies demonstrate the predictive and explanatory power of a functional approach to plant-herbivore and plant-pathogen interactions.
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  • 9
    ISSN: 1570-7458
    Keywords: ectoparasitoids ; semiochemicals ; parasite host-selection ; Eupelmus vuilleti ; Hymenoptera ; Eupelmidae ; Dinarmus basalis ; Pteromalidae ; Coleoptera ; Bruchidae ; Bruchidius atrolineatus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
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  • 10
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    Entomologia experimentalis et applicata 80 (1996), S. 311-314 
    ISSN: 1570-7458
    Keywords: leaf beetles ; Coleoptera ; Chrysomelidae ; feeding preference ; mating ; willows ; Salix
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
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  • 11
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    Entomologia experimentalis et applicata 81 (1996), S. 47-51 
    ISSN: 1570-7458
    Keywords: southern corn rootworm ; plant-insect interaction ; plant chemistry ; Coleoptera ; Chrysomelidae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The effects of host plant and dietary cucurbitacin on the growth of larval southern corn rootworm (SCR), Diabrotica undecimpunctata howardi Barber (Chrysomelidae: Luperini), were investigated. SCR were reared on four hosts: corn, Zea mays; peanuts, Arachis hypogaea; and two squash varieties, Cucurbita pepo cv. Ambassador (containing cucurbitacin D (0.08 mg g−1 fr.wt.) = bitter), and C. pepo cv. Early Yellow Crookneck (lacking cucurbitacin = non-bitter). Larval growth was significantly greater on corn and peanuts than on either squash variety. After four weeks, adults had emerged from corn and peanut plants, while squash-reared larvae had not yet entered the pupal stage. There was no difference in larval growth on the two varieties of squash. Primary metabolite measurements showed no nutritional differences between the two squash varieties. Artificial diet experiments were used to test the effect of three concentrations of cucurbitacin D (0.0, 0.1, and 0.6 mg g−1 diet) on growth of larval SCR. Larvae reared on diet containing 0.6 mg g−1 cucurbitacin weighed significantly less than larvae reared on diet containing 0.1 mg g−1 or no cucurbitacin after 10 d. No significant difference in growth was measured between the 0.1 mg g−1 diet and the 0.0 mg g−1 diet. Results are discussed relative to theories about the relationship between diabroticites and cucurbitacins.
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  • 12
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    Entomologia experimentalis et applicata 79 (1996), S. 9-17 
    ISSN: 1570-7458
    Keywords: Anthonomus pomorum ; temperature ; diel cycle ; Coleoptera ; Curculionidae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Individual pairs of overwintered adult apple blossom weevils, Anthonomus pomorum (L.), confined with apple twigs under different ambient temperatures in the laboratory and on apple trees in the field, were observed through day and night for their spring activities. Flight behavior in relation to ambient temperature was also investigated under laboratory conditions using flight stands. Both sexes displayed predominantly nocturnal behavior patterns in both the laboratory and the field. Feeding, crawling, and mating activities increased following sunset in the field or onset of scotophase in the laboratory while resting occurred most frequently during daylight hours. Results of the laboratory experiments showed that temperature affected significantly the activity patterns. The diel pattern of activities became less distinctive at higher temperatures (above 15°C), and total activities in crawling, feeding, and mating were suppressed significantly at lower temperatures (below 5°C). Over 97% of the test weevils initiated take-off response from flight stands at 20°C within the 30 min trial period; however, flight initiation rarely occurred at temperatures 12°C or below. Overall, results of the laboratory and field experiments indicate that A. pomorum is a remarkably cold-adapted insect with ability to crawl, feed, and mate at a few degrees above freezing, a physiological attribute necessary for the exploitation of early stages of apple bud development in the cold early spring.
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  • 13
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    Entomologia experimentalis et applicata 80 (1996), S. 409-414 
    ISSN: 1570-7458
    Keywords: Coleoptera ; Scarabaeidae ; Antitrogus consanguineus ; Antitrogus parvulus ; avidin ; snowdrop lectin ; wheatgerm lectin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Snowdrop and wheatgerm lectins were found to be insecticidal and growth inhibiting dietary proteins for larvae of the sugarcane whitegrub Antitrogus parvulus. At concentrations as low as 0.5 mg of snowdrop lectin per gram of semi-artificial diet, growth was inhibited by 21 days of feeding and significant mortality was apparent by 28 days. Wheatgerm lectin was active at similar concentrations, although expression of the effects was slower. Avidin was found to be a growth inhibiting dietary protein for larvae of Antitrogus consanguineus. At levels as low as 0.01 mg g-1 of diet, growth was inhibited by 28 days of feeding. Avidin caused no significant mortality after 35 days of feeding. Snowdrop and wheatgerm lectins and avidin are insect growth-inhibiting proteins whose genes potentially could be manipulated into sugarcane and improve host-plant resistance to whitegrubs.
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  • 14
    ISSN: 1572-8889
    Keywords: Prostephanus truncatus ; larger grain borer ; Coleoptera ; Bostrichidae ; flight ; stored product insect
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Higher flight activity has been observed in aged, high-density cultures ofProstephanus truncatus (Horn) (Coleoptera: Bostrichidae), but adults in new, lowdensity culture jars showed less flight activity. In order to understand this change in behavior, the effects of population density, age, resource quality, and sex on the flight ofP. truncatus were studied in a wind tunnel. While an immediate density on the release platform had no significant effect on flight, beetles from high-density cultures were more inclined to fly than those from low-density cultures. Resource quality exerted a major influence on flight; insects in food suitable for boring and oviposition seldomly exhibited flight, however, when food was absent or of inferior quality for boring and oviposition, the dominant behavior was flight. Also, insects maintained for a week in food suitable for boring and oviposition were less ready to fly than those maintained in food unsuitable for boring and oviposition. The optimum age range for flight activity was before the peak of reproduction and insects rarely flew before 4 days or after 32 days of emergence. There were no significant differences between the flight activity of males and that of females. Based on these results, we conclude that age and resource quality are major influences on the flight activity ofP. truncatus and a hypothesis is proposed in which reproductively active male and female beetles disperse from habitats of low resource quality to those that support their reproductive behavior. The practical implications of these results and the possible role of the male-produced aggregation pheromone are discussed.
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  • 15
    ISSN: 1572-8773
    Keywords: EPR ; Saccharomyces cerevisiae ; uptake ; vanadate ; vanadyl
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Vanadium uptake by whole cells and isolated cell walls of the yeast Saccharomyces cerevisiae was studied. When orthovanadate was added to wild-type S. cerevisiae cells growing in rich medium, growth was inhibited as a function of the VO4 3- concentration and the growth was completely arrested at a concentration of 20 mM of VO4 3- in YEPD. Electron paramagnetic resonance (EPR) spectroscopy was used to obtain structural and dynamic information about the cell-associated paramagnetic vanadyl ion. The presence of EPR signals indicated that vanadate was reduced by whole cells to the vanadyl ion. On the contrary, no EPR signals were detected after interaction of vanadate with isolated cell walls. A ‘mobile’ and an ‘immobile’ species associated in cells with small chelates and with macromolecular sites, respectively, were identified. The value of rotational correlation time τ r indicated the relative motional freedom at the macromolecular site. A strongly ‘immobilized’ vanadyl species bound to polar sites mainly through coulombic attractions was detected after interaction of VO2+ ions with isolated cell walls.
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  • 16
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    Entomologia experimentalis et applicata 68 (1993), S. 9-13 
    ISSN: 1570-7458
    Keywords: bioassay ; behavioural response ; Coleoptera ; carob extract ; stored product
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The behavioural responses ofCryptolestes pusillus Oryzaephilus surinamensis andProstephanus truncatus to different doses of carob extract were assessed in a two-choice pitfall bioassay and in plastic grain probe traps. The extract evoked a quick directional response and induced high beetle attraction to treated pitfall arenas compared with the controls. Overall, the extract improved probe trap efficiency by about 50% compared to unbaited probes. Beetle response was dose-dependent with the medium dose of 10 μl probably eliciting optimum response.
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  • 17
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    Cellular and molecular life sciences 49 (1993), S. 171-174 
    ISSN: 1420-9071
    Keywords: Coleoptera ; Meloidae ; Epicauta funebris ; chemical defense ; biosynthesis ; terpenoid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Cantharidin, a potent defensive chemical, is present in all ten life stages of the blister beetleEpicauta funebris. The first five larval stages accumulate cantharidin as they feed and grow in size. When disturbed, they exude cantharidin in a milky oral fluid, not in hemolymph which adult beetles reflexively discharge from leg joints. Two subsequent larval stages and the pupa do not feed, grow, regurgitate, or change in their defensive reserves (110 μg cantharidin/insect, regardless of sex). Adult beetles kept in isolation for 60–90 d exhibit a pronounced sexual dimorphism in cantharidin production: the male biosynthesizes about 17 mg of the toxin, representing 10% of his live weight, whereas the female actually loses most of her defensive reserves. But in the wild a female beetle repeatedly acquires cantharidin as copulatory gifts from her mates.
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  • 18
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    Cellular and molecular life sciences 49 (1993), S. 272-275 
    ISSN: 1420-9071
    Keywords: Semiochemical ; pheromone ; host selection ; competition ; Coleoptera ; Scolytidae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Bark beetles,Ips typographus andPityogenes chalcographus, attracted by synthetic or natural pheromone to Norway spruce logs,Picea abies, preferred to colonize uninfested logs rather than logs occupied by these beetles, probably as a means of avoiding intra-and interspecific competition. The aggregation pheromone components ofP. chalcographus, chalcogran and methyl (E, Z)-2,4-decadienoate, inhibited the attraction response ofI. typographus to its pheromone components (methyl butenol andcis-verbenol), while the converse was not true. However, verbenone released from colonized bark inhibited pheromonal response ofP. chalcographus.
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  • 19
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    Cellular and molecular life sciences 52 (1996), S. 1130-1135 
    ISSN: 1420-9071
    Keywords: Saccharomyces cerevisiae ; mitochondria ; mRNA-specific translational activation ; synthetic genes ; gene regulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Mitochondrial gene expression in yeast,Saccharomyces cerevisiae, depends on translational activation of individual mRNAs by distinct proteins encoded in the nucleus. These nuclearly coded mRNA-specific translational activators are bound to the inner membrane and function to mediate the interaction between mRNAs and mitochondrial ribosomes. This complex system, found to date only in organelles, appears to be an adaptation for targeting the synthesis of mitochondrially coded integral membrane proteins to the membrane. In addition, mRNA-specific translational activation is a rate-limiting step used to modulate expression of at least one mitochondrial gene in response to environmental conditions. Direct study of mitochondrial gene regulation and the targeting of mitochondrially coded proteins in vivo will now be possible using synthetic genes inserted into mtDNA that encode soluble reporter/passenger proteins.
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  • 20
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    Cellular and molecular life sciences 52 (1996), S. 1033-1041 
    ISSN: 1420-9071
    Keywords: Ubiquitin ; yeast ; Saccharomyces cerevisiae ; Dictyostelium discoideum ; cytoskeleton ; mutants ; endocytosis ; actin ; myosin ; calmodulin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Endocytosis is a general term that is used to describe the internalization of external and plasma membrane molecules into the cell interior. In fact, several different mechanisms exist for the internalization step of this process. In this review we emphasize the work on the actin-dependent pathways, in particular in the yeastSaccharomyces cerevisiae, because several components of the molecular machinery are identified. In this yeast, the analysis of endocytosis in various mutants reveals a requirement for actin, calmodulin, a type I myosin, as well as a number of other proteins that affect actin dynamics. Some of these proteins have homology to proteins in animal cells that are believed to be involved in endocytosis. In addition, the demonstration that ubiquitination of some cell surface molecules is required for their efficient internalization is described. We compare the actin, myosin and ubiquitin requirements for endocytosis with recent results found studying these processes usingDictyostelium discoideum and animal cells.
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  • 21
    ISSN: 1420-9071
    Keywords: Coleoptera ; Coccinellidae ; Subcoccinella-24-punctata ; chemical defense ; quinoline alkaloid ; Nα-quinaldyl-L-arginine·HCl
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The isolation of Nα-quinaldyl-L-arginine·HCl (1) from the CoccinellidaeSubcoccinella-24-punctata is reported. The structure, first established on the basis of the analysis of the spectral properties of1, has been confirmed by synthesis. The alkaloid is of endogenous origin and markedly deterrent to ants.
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  • 22
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    Cellular and molecular life sciences 52 (1996), S. 1111-1116 
    ISSN: 1420-9071
    Keywords: Mitochondria ; mitochondrial inheritance ; cytoskeleton ; Saccharomyces cerevisiae ; Schizosaccharomyces pombe ; membrane proteins ; organelle movement ; mitochondrial morphology
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Mechanisms mediating the inheritance of mitochondria are poorly understood, but recent studies with the yeastsSaccharomyces cerevisiae andSchizosaccharomyces pombe have begun to identify components that facilitate this essential process. These components have been identified through the analysis of conditional yeast mutants that display aberrant mitochondrial distribution at restrictive conditions. The analysis of these mutants has uncovered several novel proteins that are localized either to cytoskeletal structures or to the mitochondria themselves. Many mitochondrial inheritance mutants also show altered mitochondrial morphology and defects in maintenance of the mitochondrial genome. Although some inheritance components and mechanisms appear to function specifically in certain types of cells, other conserved proteins are likely to mediate mitochondrial behavior in all eukaryotic cells.
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  • 23
    ISSN: 1420-9071
    Keywords: Saccharomyces cerevisiae ; mitochondrial ribosomes ; peptidyl transferase ; Varl ribosomal protein ; gene relocation ; posttranscriptional rRNA modification
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Mitochondria posses their own ribosomes responsible for the synthesis of a small number of proteins encoded by the mitochondrial genome. In yeast,Saccharomyces cerevisiae, the two ribosomal RNAs and a single ribosomal protein, Varl, are products of mitochondrial genes, and the remaining approximately 80 ribosomal proteins are encoded in the nucleus. The mitochondrial translation system is dispensable in yeast, providing an excellent experimental model for the molecular genetic analysis of the fundamental properties of ribosomes in general as well as adaptations required for the specialized role of ribosomes in mitochondria. Recent studies of the peptidyl transferase center, one of the most highly conserved functional centers of the ribosome, and the Varl protein, an unusual yet essential protein in the small ribosomal subunit, have provided new insight into conserved and divergent features of the mitochondrial ribosome.
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  • 24
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    Chemoecology 4 (1993), S. 29-32 
    ISSN: 1423-0445
    Keywords: chemical defence ; alkaloids ; predation ; Coleoptera ; Coccinellidae ; Adalia bipunctata ; Coccinella septempunctata ; Hymenoptera ; Formicidae ; Lasius niger
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Garden black ants,Lasius niger L., in a laboratory colony, attacked three species of live ladybirds found near their nest, killing the smaller two species. A second colony was offered artificial diets containing crushed ladybirds of two species, and the ants' choice of feeding site noted. Both the diets were aversive compared to control, but that containing 7spot,Coccinella septempunctata L., was more aversive than the diet containing 2spot,Adalia bipunctata L. The implications of this lesser protection for 2spots in terms of the chemical defence of the species are discussed.
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  • 25
    ISSN: 1423-0445
    Keywords: herbivory ; plant resistance ; leaf colour ; leaf toughness ; tree growth rate ; leaf nutrients ; terpenes ; Coleoptera ; Chrysomelidae ; Paropsinae ; Chrysophtharta bimaculata ; Myrtaceae ; Eucalyptus regnans
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In bagged and unbagged shoot experiments, we investigated the survival and growth rate of first instar larvae ofChrysophtharta bimaculata on 9 families of a natural host,Eucalyptus regnans. Families used had been previously assessed as being either of low or high susceptibility toC. bimaculata damage. In conjunction with larval experiments, we measured 24 tree and leaf characteristics (including foliar elemental concentrations, foliar terpenes, leaf toughness and tree growth rates) and attempted to correlate the plant characters measured with differences in larval performance and previous scorings ofE. regnans family susceptibility. First instar larval growth and survival did not differ significantly across families or between low and high susceptibility family groups (=susceptibility classes), although survival was significantly greater in bagged than unbagged treatments. As predators were in low abundance at the study site, we attribute higher survival rates of larvae in bagged treatments to increased protection from adverse weather conditions. Only one plant character measured, an unidentified foliar phlorglucinol, was significantly negatively correlated with larval survival. Of the 24 plant characters measured, 11 were significantly different between families and 10 were signficantly different between susceptibility classes. Only 4 plant characters were significantly different at both the family and susceptibility class levels;viz. proportion red leaves, tree height at end of season, trunk volume at end of season and relative growth rate based on tree height. Principle Component Analysis using all plant characters measured, or subsets of them, could not separate individual families or susceptibility classes. Our results suggest that herbivore resistance mechanisms inE. regnans do not affectC. bimaculata larvae, but may influence adult feeding and/or oviposition.
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  • 26
    ISSN: 1423-0445
    Keywords: male aggression ; defensive secretion ; tergal gland ; closed-loop-stripping-analyses ; tandem bioassay ; mass fragmentography ; 2-methyl-1,4-benzoquinone ; Coleoptera ; Staphylinidae ; Aleochara curtula
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary 2-methyl-1,4-benzoquinone as a target compound of the tergal gland secretion of the rove beetle,Aleochara curtula, was quantified from the reservoirs of individual beetles. Males store less secretion than females, and they evaporate more of the secretion as measured by adsorption from the air (closed-loop-stripping-analysis). The amount of emitted quinone is increased during aggressive interactions of rival males. The pulsed emission of the secretion during aggression is demonstrated by a novel technique combining observation of behavior with the on-line measurement of target fragment ions by mass-spectrometry (tandem bioassay — mass fragmentography). The emission of the secretion is used as a weapon in combats between males and may result in the repulsion of subordinate males from the mating site, but may also serve to mimic females chemically in order to avoid aggressive encounters.
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  • 27
    ISSN: 1423-0445
    Keywords: chemical defense ; predator-prey interaction ; synergism ; Coleoptera ; Staphylinidae ; Oxytelinae ; Bledius ; Carabidae ; Dyschirius ; Pogonus ; Dichirotrichus ; Formicinae ; Cataglyphis ; Dermaptera ; Labidura
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The adaptation of defensive secretions to their target organisms was examined for the abdominal gland secretions ofBledius furcatus, B. spectabilis andB. arenarius. Therefore the target organisms of the secretion of theseBledius species (i.e. their predators) had to be identified. At the collection sites examined these were the earwigLabidura riparia, the antCataglyphis bicolor, the flyLispe candicans, different carabids of the generaPogonus, Dichirotrichus, Dyschirius, Bembidion andCalathus and the wading birdsHaematopus ostralegus andCalidris alba. The secretion of the abdominal glands contains the toxin ptoluquinone dissolved in eitherγ-dodecalactone and 1-undecene (B. furcatus andB. spectabilis) or in octanoic acid and octyloctanoate (B. arenarius). The ratio of these solvents is species-specific. Application experiments using some of the natural insect predators (L. riparia, C. bicolor, Pogonus, Di. gustavii, Dyschirius) revealed that these solvent ratios provided a more effective deterrent than other possible ratios. Thus by combining the solvents in certain ratios, the capability of cuticular penetration and therefore the effectiveness of the defensive secretions are adapted to their natural targets.
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  • 28
    ISSN: 1423-0445
    Keywords: kairomone ; behavior ; cucurbitacin ; Cucurbitaceae ; Coleoptera ; Chrysomelidae ; Luperini ; Aulacophora ; Diabrotica
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The attraction of Old World leaf beetles in the genusAulacophora to kairomones and parakairomones which are effective lures for New WorldDiabrotica andAcalymma were investigated. Beetles captured on sticky traps baited with single and multicomponent lures were no different from the control traps for two species ofAulacophora. Yellow colored traps and squash blossoms are attractive toAulacophora beetles which detect sub-microgram quantities of cucurbitacins on silica gel. Leaf feeding behavior and flight activity data are correlated with varietal preference of threeAulacophora species. The common response byDiabrotica andAulacophora to cucurbitacins reinforces the two groups' coevolutionary association with the Cucurbitaceae. The apparent lack of a common response toCucurbita blossom volatiles suggests recent evolutionary pathways are substantially different for these two groups of beetles.
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  • 29
    ISSN: 1423-0445
    Keywords: subsociality ; cleptoparasitism ; mimetism ; odour congruity ; homology ; defensive secretions ; Kalahari desert ; Coleoptera ; Tenebrionidae ; Parastizopus armaticeps ; Eremostibes opacus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Parastizopus armaticeps andEremostibes opacus are two closely related desert tenebrionids which also live in close association, the former having biparental brood care and the latter cleptoparasitising the brood.E. opacus is unable to discriminate between the conspecific and host odour even in the absence of physical contact. Gas chromatographic analysis of headspace volatiles of resting animals showed almost complete qualitative and quantitative odour congruity between them. Comparison of these odour profiles with those of two other tenebrionids sharing the same ecological niche,Gonopus agrestis andHerpiscius sp. (damaralis?) showed that congruity was independent of common foodplant utilisation. It is also independent of common defensive gland secretions. Parallels between resting odour spectra, defensive secretion spectra and systematic status suggest that the origins of congruity lie in odour homology, by means of which the cleptoparasite was able to exploit its host.
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  • 30
    ISSN: 1423-0445
    Keywords: volatile substances ; pygidial secretion ; defence ; communication ; species differences ; Coleoptera ; Gyrinidae ; Gyrinus spp
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Gyrinid beetles are common in freshwater habitats. They have paired pygidial glands with a secretion that contains high molecular weight substances rendering them toxic for predatory fish. In this paper we report on a laboratory study on volatile components released by three different gyrinid species when irritated. The volatile pattern had a clear difference between the different species.Gyrinus substriatus andG. aeratus, both produced 3-methyl-1-butanal and 3-methyl-1-butanol when irritated, but in quite different amounts. The third tested species,G. minutus, did not produce any substances above the detection level. It is suggested that the volatile compounds may be part of the beetles' communication and/or defence system.
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  • 31
    ISSN: 1432-1939
    Keywords: Coleoptera ; Herbivory ; Host selection ; Maternal effect ; Paternal effect
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The natural host of Ophraella notulata is Iva frutescens (Asteraceae); its close relative feeds on a related plant, Ambrosia artemisiifolia. We reared beetles on both plants, obtained progeny from the four possible crosses (two sexes X two parental hosts), and reared the progeny on both plant species. Survival to the imaginal stage of progeny reared on Iva varied with both maternal and paternal host. Hatchling feeding response to both plants showed a maternal host X paternal host interaction. Consumption of Ambrosia by adult beetles was, counter to expectation, higher for progeny of Iva-reared males than Ambrosia-reared males. Oviposition response, although based on too few data to be definitive, was peculiar: parental host did not affect oviposition on Ambrosia; on Iva daughters of Iva-reared males laid significantly more eggs than did daughters of Ambrosia-reared males, but only if they had been reared on Iva; those reared on Ambrosia displayed the reverse pattern. We discuss the possibility that nongenetic paternal transmission of host plant effects may explain these results, but offer a somewhat uncomfortable hypothesis of selection as a preferable explanation. An important outcome of the experiment is that it provided no evidence of maternal effects of host plant on offspring feeding or oviposition.
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  • 32
    ISSN: 1432-0983
    Keywords: Glucoamylase ; Gene cloning ; Hormoconis resinae ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A cDNA coding for glucoamylase P of Hormoconis resinae was cloned using a synthetic oligonucleotide probe coding for a peptide fragment of the purified enzyme and polyclonal anti-glucoamylase antibodies. Nucleotide-sequence analysis revealed an open reading frame of 1848 base pairs coding for a protein of 616 amino-acid residues. Comparison with other fungal glucoamylase amino-acid sequences showed homologies of 37–48%. The glucoamylase cDNA, when introduced into Saccharomyces cerevisiae under the control of the yeast ADC1 promoter, directed the secretion of active glucoamylase P into the growth medium.
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  • 33
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Dynamin ; Mitochondria ; GTP binding protein
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The isolation and characterization of MGM1, and yeast gene with homology to members of the dynamin gene family, is described. The MGM1 gene is located on the right arm of chromosome XV between STE4 and PTP2. Sequence analysis revealed a single open reading frame of 902 residues capable of encoding a protein with an approximate molecular mass of 101 kDa. Loss of MGM1 resulted in slow growth on rich medium, failure to grow on non-fermentable carbon sources, and loss of mitochondrial DNA. The mitochondria also appeared abnormal when visualized with an antibody to a mitochondrial-matrix marker. MGM1 encodes a dynamin-like protein involved in the propagation of functional mitochondria in yeast.
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  • 34
    ISSN: 1432-0983
    Keywords: Growth control ; Genetic mapping ; Molecular cloning ; Nucleo-mitochondrial interaction ; Saccharomyces cerevisiae ; Viability of petites
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The PEL1 gene of Saccharomyces cerevisiae is essential for the cell viability of mitochondrial petite mutants, for the ability to utilize glycerol and ethanol on synthetic medium, and for cell growth at higher temperatures. By tetrad analysis the gene was assigned to chromosome III, centromere proximal of LEU2. The PEL1 gene has been isolated and cloned by the complementation of a pel1 mutation. The molecular analysis of the chromosomal insert carrying PEL1 revealed that this gene corresponds to the YCL4W open reading frame on the complete DNA sequence of chromosome III. The putative Pel1 protein is characterized by a low molecular weight of approximately 17 kDa, a low codon adaptation index, and a high leucine content.
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  • 35
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Papaver somniferum L. ; ARS ; Mitochondrial DNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The minimal fragment of mitochondrial DNA from Papaver somniferum L. (poppy) able to promote autonomous plasmid replication in the yeast Saccharomyces cerevisiae was sequenced. Sequence analysis of the 917-bp MK4/8 DNA fragment revealed a high AT content, and the presence of two 12-bp sequences differing from the ARS core consensus of S. cerevisiae only by a T and C insertion, respectively. The mitochondrial insert contains a further six 11-bp sequences with one mismatch to the S. cerevisiae core consensus, more then 20 related sequences with two base pair exchanges, numerous direct and inverted repeats, and many copies of a sequence motif called the ARS box. The original 4.2-kb mitochondrial DNA fragment, as well as the minimal 917-bp subfragment in vector pFL1-E (a variant of YIP5, lacking an origin of replication in yeast), were then tested for their ability to replicate autonomously in another fungus, Kluyveromyces lactis.
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  • 36
    ISSN: 1432-0983
    Keywords: 2-Oxoglutarate dehydrogenase ; Molecular cloning ; Saccharomyces cerevisiae ; Sequencing ; Suppressor ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The activity of mitochondrial 2-oxoglutarate dehydrogenase in S. cerevisiae can be impaired either by the ogd1 or the kgd1 mutation. The OGD1 gene and two suppressor genes were isolated by complementation of the ogd1 mutant. The complementation of the kdg1 mutant by the OGD1 gene, an allelism test, and meiotic mapping, revealed that the ogd1 and kgd1 mutations are allelic. The two mutations were differentiated by the cloned suppressor gene which was able to partially complement ogd1, but not kgd1. The molecular analysis of the suppressor gene revealed its identity with the natural tRNA CAG Gln gene found in the upstream region of URA10.
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  • 37
    ISSN: 1432-0983
    Keywords: Yeast ; Saccharomyces cerevisiae ; Sporulation mutants ; Reporter genes
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    Topics: Biology
    Notes: Abstract Reporter genes consisting of sporulation-specific promoters fused to lacZ were used as markers to monitor the sporulation pathway of the yeast Saccharomyces cerevisiae. Strains transformed with these lacZ gene fusions expressed β-galactosidase (assayable on plates using the substrate 5-bromo-4-chloro-3-indolyl-β-D-galactopyranoside, X-gal) in a sporulation-dependent manner. Mutagenesis experiments performed on transformed strains resulted in the recovery of a number of novel sporulation mutants. Three classes of mutants were obtained: those which overexpressed the reporter gene under sporulation conditions, those which did not express the gene under any conditions, and those which expressed the gene in vegetative cells not undergoing sporulation. On the basis of the blue colony-colour produced in the presence of X-gal these have been described as superblue, white, and blue vegetative mutants, respectively. These were further characterised using earlier reporter genes and other marker systems. This study established that the multicopy reporter plasmids chosen do not interfere with sporulation; they are valid tools for monitoring the pathway and they provide a way to isolate mutations not readily selected by other markers.
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  • 38
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    Current genetics 24 (1993), S. 461-464 
    ISSN: 1432-0983
    Keywords: Chromosome fragmentation ; MEL gene family ; Saccharomyces cerevisiae
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    Topics: Biology
    Notes: Abstract Nine members, MEL2–MEL10, of the MEL gene family coding for α-galactosidase were physically mapped to the ends of the chromosomes by chromosome fragmentation. Genetic mapping of the genes supported the location of all the MEL genes in the left arm of their resident chromosomes.
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  • 39
    ISSN: 1432-0983
    Keywords: Yeast ; Saccharomyces cerevisiae ; Transformation ; Plasmid
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    Notes: Abstract We have compared a number of procedures for the transformation of whole cells of the yeast Saccharomyces cerevisiae and assessed the effects of dimethylsulphoxide (DMSO) or ethanol, both of which have been reported to enhance transformation efficiency. We find that simplified methods benefit from the addition of one of these compounds, and although differences are observed between strains as to the more beneficial reagent, peak transformation efficiency is, in general obtained with 10% DMSO or 10% EtOH. Increases of between six- and 50-fold are observed, despite a reduction in cell viability, and at this concentration the two compounds are not additive in their effects. The optimum level appears to depend on a balance between improved DNA uptake and reduced cell viability. As a result of this work we present a straightforward and rapid transformation procedure.
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  • 40
    ISSN: 1432-0983
    Keywords: Key words Omnipotent suppression ; Microtubules ; Respiratory deficiency ; Saccharomyces cerevisiae
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    Notes: Abstract  SUP35 and SUP45 genes determine the accuracy of translation at the stage of termination. We present indirect evidence indicating that these genes may also control some cellular process mediated by microtubules. A majority of sup35 and sup45 suppressor mutations confer supersensitivity to benomyl, the drug which de-polymerizes microtubules. In addition, data correlating phenotypic manifestations of sup45 suppressor mutations, involving sensitivity to benomyl, respiratory deficiency and a suppressor effect, are also presented.
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  • 41
    ISSN: 1432-0983
    Keywords: Key words D-ribulose-5-phosphate 3-epimerase ; D-ribose-5-phosphate ketol-isomerase ; Pentose-phosphate pathway ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have cloned and characterized the two remaining unknown genes of the non-oxidative part of the pentose-phosphate pathway of Saccharomyces cerevisiae encoding the enzymes D-ribulose-5-phosphate 3-epimerase (Rpe1p) and D-ribose-5-phosphate ketol-isomerase (Rki1p). Rpe1p has an unexpected high specific activity of 2148 mU × (mg protein)–1 in crude extracts. Deletion mutants of RPE1 show no enzyme activity and are unable to grow on D-xylulose. Unexpectedly, haploid rki1 deletion mutants are not viable. Functional expression of RKI1 was demonstrated following an increase of gene dosage in the haploid rki1 deletion mutant, which restored viability and specific D-ribose-5-phosphate ketol-isomerase activity. Both enzymes show high similarity to the deduced protein sequences of various open reading frames, expressed sequence tags or cDNAs from different organisms.
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  • 42
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    Current genetics 30 (1996), S. 461-468 
    ISSN: 1432-0983
    Keywords: Keywords DNA repair ; Methylation damage ; Epistasis analysis ; Saccharomyces cerevisiae
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    Topics: Biology
    Notes: Abstract  The major genotoxicity of methyl methanesulfonate (MMS) is due to the production of a lethal 3-methyladenine (3MeA) lesion. An alkylation-specific base-excision repair pathway in yeast is initiated by a Mag1 3MeA DNA glycosylase that removes the damaged base, followed by an Apn1 apurinic/ apyrimidinic endonuclease that cleaves the DNA strand at the abasic site for subsequent repair. MMS is also regarded as a radiomimetic agent, since a number of DNA radiation-repair mutants are also sensitive to MMS. To understand how these radiation-repair genes are involved in DNA methylation repair, we performed an epistatic analysis by combining yeast mag1 and apn1 mutations with mutations involved in each of the RAD3, RAD6 and RAD52 groups. We found that cells carrying rad6, rad18, rad50 and rad52 single mutations are far more sensitive to killing by MMS than the mag1 mutant, that double mutants were much more sensitive than either of the corresponding single mutants, and that the effects of the double mutants were either additive or synergistic, suggesting that post-replication and recombination-repair pathways recognize either the same lesions as MAG1 and APN1, or else some differ- ent lesions produced by MMS treatment. Lesions handled by recombination and post replication repair are not simply 3MeA, since over-expression of the MAG1 gene does not offset the loss of these pathways. Based on the above analyses, we discuss possible mechanisms for the repair of methylation damage by various pathways.
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  • 43
    ISSN: 1432-0983
    Keywords: Biocontrol ; Secretion ; Chitinase ; Expression cloning ; Saccharomyces cerevisiae ; Trichoderma harzianum
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    Topics: Biology
    Notes: Abstract A novel endochitinase agar-plate assay has been developed and used to identify 11 full-length cDNAs encoding endochitinase I (ENC I) from aTrichoderma harzianum cDNA library by expression in yeast. The 1473-bpchil cDNA encodes a 424-residue precursor protein including both a signal sequence and a propeptide. The deduced ENC I amino-acid sequence is homologous to other fungal and bacterial chitinases, and the enzyme cross-reacts with a polyclonal antiserum raised against chitinase A1 fromBacillus circulans. TheT. harzianum endochitinase I was secreted into the culture medium by the yeastSaccharomyces cerevisiae in a functionally active form. The purified recombinant enzyme had a molecular mass of 44 kDa, an isoelectric point of 6.3, a pH optimum of 7.0 and a temperature optimum of 20 °C.
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  • 44
    ISSN: 1432-0983
    Keywords: Heat-shock response ; Multidrug resistance ; AP-1 homolog ; Saccharomyces cerevisiae
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    Topics: Biology
    Notes: Abstract We have examined whether the stress-induced transcriptional activation ofYDR1/PDR5/STS1 is mediated by yAP-1 and yAP-2. Of the stresses examined, heat shock-induced, rapid and transient PDR5 expression became very low in ayap1 yap2 double-gene disruptant, indicating that the yAP proteins mediate the response. Similar results were obtained withSNQ2, a close homologue ofPDR5. A set of 5′-truncation derivatives of thePDR5 gene identified the region from −484 to −434 as being sufficient for the response. A sequence similar to the yAP-1 recognition element recently identified in the stress-responsive yeast genes was found in this region and in the 5′-flanking sequences ofSNQ2.
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  • 45
    ISSN: 1432-0983
    Keywords: Key words Biocontrol ; Secretion ; Chitinase ; Expression cloning ; Saccharomyces cerevisiae ; Trichoderma harzianum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  A novel endochitinase agar-plate assay has been developed and used to identify 11 full-length cDNAs encoding endochitinase I (ENC I) from a Trichoderma harzianum cDNA library by expression in yeast. The 1473-bp chi1 cDNA encodes a 424-residue precursor protein including both a signal sequence and a propeptide. The deduced ENC I amino-acid sequence is homologous to other fungal and bacterial chitinases, and the enzyme cross-reacts with a polyclonal antiserum raised against chitinase A1 from Bacillus circulans. The T. harzianum endochitinase I was secreted into the culture medium by the yeast Saccharomyces cerevisiae in a functionally active form. The purified recombinant enzyme had a molecular mass of 44 kDa, an isoelectric point of 6.3, a pH optimum of 7.0 and a temperature optimum of 20 °C.
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  • 46
    ISSN: 1432-0983
    Keywords: Glycosylphosphatidylinositol anchored-protein ; Southern analysis ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The GGP1 gene encodes the only GPI-anchored glycoprotein (gp115) that has been purified todate in the budding yeast Saccharomyces cerevisiae. It is a single-copy gene whose deduced amino-acid sequence shares no significant homology to any other known protein. In this paper we report a Southern hybridization analysis of genomic DNA from different eukaryotic organisms to identify homologues of the GGP1 gene. We have analyzed DNA prepared from a unicellular green alga (Chlamydomonas eugametos), from two distantly related yeast species (Candida cylindracea and Schizosaccharomyces pombe), and from the common bean Phasoleus vulgaris. The moderate stringency of the experimental conditions and the high specificity of the probes used indicate that a single-copy of GGP1-related sequences exists in all these eukaryotic organisms. The chromosomal localization of the GGP1 gene in S. cerevisiae has also been determined.
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  • 47
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    Current genetics 23 (1993), S. 92-94 
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Gene mapping ; Idiomorphism
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The STA2 (glucoamylase) gene of Saccharomyces cerevisiae has been mapped close to the end of the left arm of chromosome II. Meiotic analysis of a cross between a haploid strain containing STA2, and another strain carrying the melibiase gene MEL1 (which is known to be at the end of the left arm of chromosome II) produced parental ditype tetrads only. Since there is no significant DNA sequence similarity between the STA2 and MEL1 genes, or their respective flanking regions, we conclude that these two genes are carried by separate non-hybridizing sequences of chromosomal DNA, either of which can reside at the end of the left arm of chromosome II. By analogy with the mating-type locus of Neurospora crassa, we suggest that the STA2 and MEL1 genes are idiomorphs with respect to one another.
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  • 48
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Phospholipid synthesis ; Phospholipid-N-methyltransferase ; Mutant ; Over-expression
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    Topics: Biology
    Notes: Abstract By functional complementation of the auxotrophic requirements for choline of a cdg1, cho2 double-mutant, by transformation with a genomic DNA library in a high copy number plasmid, two different types of complementing DNA inserts were identified. One type of insert was earlier shown to represent the CHO2 structural gene. In this report we describe the molecular and biochemical characterization of the second type of complementing activity. The transcript encoded by the cloned gene was about 1000-nt in length and was regulated in response to the soluble phospholipid precursors, inositol and choline. A gene disruption resulted in no obvious growth phenotype at 23°C or 30°C, but in a lack of growth at 37°C in the presence of monomethylethanolamine. Null-mutants exhibited an inositol-secretion phenotype, indicative of mutations in the lipid biosynthetic pathway. Complementation analysis, biochemical analysis of the phospholipid methylation pathway in vivo, and comparison of the restriction pattern of the cloned gene to published sequences, unequivocally identified the cloned gene as the OPI3 gene, encoding phospholipid-N-methyltransferase in yeast. When present in multiple copies the OPI3 gene efficiently suppresses the phospholipid methylation defect of a cho2 mutation. As a result of impaired synthesis of phosphatidylcholine, the INO1-deregulation phenotype is abolished in cho2 mutants transformed with the OPI3 gene on a high copy number plasmid. Taken together, these data demonstrate a significantly overlapping specificity of the OPI3 gene product for three sequential phospholipid methylation reactions in the de novo Ptd-Cho biosynthetic pathway.
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  • 49
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    Current genetics 23 (1993), S. 181-183 
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; c-myc epitope ; Fusion
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    Topics: Biology
    Notes: Abstract In order to facilitate the process of epitope-tagging of yeast proteins, we have constructed two Saccharomyces cerevisiae-Escherichia coli shuttle vectors that allow fusion of a sequence encoding an epitope of the human c-myc protein at the 3′ end of any gene. An example of the use of this technique is presented.
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  • 50
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    Current genetics 23 (1993), S. 295-304 
    ISSN: 1432-0983
    Keywords: Meiosis ; Meiotic recombination ; Saccharomyces cerevisiae ; REC114
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Four new meiotic recombination genes were previously isolated by selecting for mutations that rescue the meiotic lethality of rad52 spo13 strains. One of these genes, REC114, is described here, and the data confirm that REC114 is a meiosis-specific recombination gene with no detectable function in mitosis. REC114 is located on chromosome XIII approximately 4,9 cM from CIN4. The nucleotide sequence reveals an open reading frame of 1262 bp, consensus intron splice sites close to the 3′ end, and indicates that the second exon codes for only seven amino acids. In the promoter region, a URS1 consensus sequence (TGGGCGGCTA), identical to the URS1 found in the promoter of SPO16, is present 93 bp upstream of the translation start site. Northern-blot hybridization demonstrates that REC114 is transcribed only during meiosis and that it is not expressed in the absence of the IME1 gene product, even when IME2 is constitutively expressed.
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  • 51
    ISSN: 1432-0983
    Keywords: Trehalase ; Trehalose-6-P synthase ; cAMP mutants ; Saccharomyces cerevisiae
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    Topics: Biology
    Notes: Abstract The rise in cAMP level that follows the addition of glucose or 2,4-dinitrophenol (DNP) to stationaryphase cells of Saccharomyces cerevisiae was accompanied by a marked activation of trehalase (3-fold increase) and a concomitant deactivation of trehalose-6 phosphate synthase (50% of the basal levels). In glucose-grown exponential cells, which are deficient in glucose-induced cAMP signalling, the addition of glucose also prompted a decrease in trehalose-6 phosphate synthase, but had no effect on trehalase activity. Mutants defective in the RAS-adenylate cyclase pathway (ras1 ras2 bcy1 strain), as well as mutants containing greatly reduced protein kinase activity either cAMP-dependent (tpk w1 BCY1 strains) or cAMP-independent (tpk1 w1 bcy1 strains), were unable to show glucose- or DNP-induced trehalase activation but still displayed a clear decrease in trehalose-6 phosphate synthase activity upon addition of these compounds. These data suggest that the activity of trehalose-6 phosphate synthase, as opposed to that of trehalase, is not controlled by the cAMP signalling pathway “in vivo”. Trehalose-6 phosphate synthase was competitively inhibited by glucose (Ki=15 mM) and resulted unaffected by ATP in assays performed “in vitro”.
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  • 52
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    Current genetics 23 (1993), S. 375-381 
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Isocitrate lyase ; Gene regulation ; Ethanol induction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The ICL1 gene encoding the isocitrate lyase from Saccharomyces cerevisiae was cloned and sequenced. A reading frame of 557 amino acids showing significant similarity to isocitrate lyases from seven other species could be identified. Construction of icl1 null mutants led to growth defects on C2 carbon sources while utilization of sugars or C3 substrates remained unaffected. Using an ICL1-lacZ fusion integrated at the ICL1 locus, a more than 200-fold induction of β-galactosidase activity was observed after growth on ethanol when compared with glucose-repressed conditions. A preliminary analysis of the ICL1 upstream region identified a 364-bp fragment necessary and sufficient for this regulatory phenotype. Sequence motifs also present in the upstream regions of co-regulated genes were found within this region.
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  • 53
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Gene amplification ; ADH4 ; CUP1
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    Notes: Abstract Primary gene amplification, i.e., mutation from one gene copy to multiple gene copies per genome, is important in genomic evolution, as a means of producing anti-cancer drug resistance, and is associated with the progression of tumor malignancy. Primary amplification has not been studied in normal eukaryotic cells because amplifications are extremely rare in these cells. A system has been developed to phenotypically identify co-amplifications of the ADH4 and CUP1 genes of Saccharomyces cerevisiae and 21 independent spontaneous amplifications have been isolated.
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  • 54
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    Current genetics 23 (1993), S. 414-422 
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Donation ; Gene conversion ; Double-strand break repair ; Heteroduplex DNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have used transformation of yeast with lincarized plasmids to study the transfer of information to the unbroken chromosome during double-strand break repair. Using a strain which carried the wild-type HIS3 allele, and a linearized plasmid which carried a mutant his3 allele, we have obtained His- transformants. In these, double-strand break repair has resulted in precise transfer of genetic information from the plasmid to the chromosome. Such repair events, we suggest, are gene conversions which entail the formation of heteroduplex DNA on the (unbroken) chromosome. If this suggestion is correct, our results reflect the spatial distribution of such heteroduplex DNA. Transfer of information from the plasmid to the chromosome was obtained at a maximal frequency of 1.5% of the repair events, and showed a dependence with distance. Transformation to His- was also obtained with a 2-kbp insertion and with a deletion of 200 bp. The latter results suggest that gene conversion of large heterologies can occur via repair of a heteroduplex DNA intermediate.
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  • 55
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    Current genetics 24 (1993), S. 185-192 
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Cell cycle ; Transcription ; DNA replication
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In eukaryotic organisms, genes involved in DNA replication are often subject to some form of cell cycle control. In the yeast Saccharomyces cerevisiae, most of the DNA replication genes that have been characterized to date are regulated at the transcriptional level during G1 to S phase transition. A cis-acting element termed the MluI cell cycle box (or MCB) conveys this pattern of regulation and is common among more than 20 genes involved in DNA synthesis and repair. Recent findings indicate that the MCB element is well conserved among fungi and may play a role in controlling entry into the cell division cycle. It is evident from studies in higher systems, however, that transcriptional regulation is not the only form of control that governs the cell-cycle-dependent expression of DNA replication genes. Moreover, it is unclear why this general pattern of regulation exists for so many of these genes in various eukaryotic systems. This review summarizes recent studies of the MCB element in yeast and briefly discusses the purpose of regulating DNA replication genes in the eukaryotic cell cycle.
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  • 56
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Pentose-phosphate pathway ; Transketolase
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    Topics: Biology
    Notes: Abstract Deletion mutants for the yeast transketolase gene TKL1 were constructed by gene replacement. Transketolase activity was below the level of detection in mutant crude extracts. Transketolase protein could be detected as a single protein band of the expected size by Western-blot analysis in wild-type strains but not in the delection mutant. Deletion of TKL1 led to a reduced but distinct growth in synthetic medium without an aromatic amino-acid supplement. We also isolated double and triple mutants for transketolase (tkl1), transaldolase (tal1), and glucose 6-phosphate dehydrogenase (zwf1) by crossing the different mutants. A tal1 tkl1 double mutant grew nearly like wild-type in rich medium. Only the tkl1 zwf1 double and the tal1 tkl1 zwf1 triple mutant grew more slowly than the wild-type in rich medium. This growth defect could be partly alleviated by the addition of xylulose but not ribose. The triple mutant still grew slowly on a synthetic mineral salts medium without a supplement of aromatic amino acids. This suggests the existence of an alternative but limited source of pentose phosphates and erythrose 4-phosphate in the tkl1 zwf1 double mutants. Hybridization with low stringency showed the existence of a sequence with homology to transketolase, possibly a second gene.
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  • 57
    ISSN: 1432-0983
    Keywords: Key words Cytochrome b ; Mutants ; Mitochondria ; Saccharomyces cerevisiae
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    Topics: Biology
    Notes: Abstract The nucleotide changes present in a group of five cytochrome b mit– mutants were analyzed at the sequence level. Two single-base changes were found: one (M10-152) generated a nonsense codon in the first exon while the other (M8-181) created a missense substitution in the second exon. The other mutants all have multiple (three) substitutions that either resulted in a missense mutation in a coding region (M17-162) or else changed nucleotides in the last intron of the gene, so blocking its excision (M6-200 and M8-53). The synthesis of mitochondrial polypeptides and the steady state concentration of the complex-III subunits were examined. The Rieske protein and the core-4 and core-5 subunits were much reduced in all mutants. Consequently the overall stability of complex III is very sensitive even to amino-acid substitutions in the cytochrome b protein. Mutant M8-53 provides direct evidence for the proposed role of the P9.1 stem in the core structure of the group-I type last intron of this gene.
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  • 58
    ISSN: 1432-0983
    Keywords: Key wordsβ-glucosidase ; Candida wickerhamii ; Saccharomyces cerevisiae
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    Topics: Biology
    Notes: Abstract The yeast Candida wickerhamii exports a cell-associated β-glucosidase that is active against cellobiose and all soluble cellodextrins. Because of its unique ability to tolerate end-product inhibition by glucose, the bglB gene that encodes this enzyme was previously cloned and sequenced in this laboratory. Using several different promoters and constructs, bglB was expressed in the hosts Escherichia coli, Pichia pastoris, and Saccharomyces cerevisiae. Expression was initially performed in E. coli using either the lacZ or tac promoter. This resulted in intracellular expression of the BglB protein with the protein being rapidly fragmented. Secretion and glycosylation of active β-glucosidase was achieved with several different S. cerevisiae constructs utilizing either the adh1 or the gal1 promoter on 2-µ replicating plasmids. When either the invertase (Suc2) or the BglB secretion signal was used, BglB protein remained associated with the cell wall and appeared to be hyperglycosylated. Expression in P. pastoris was also examined to determine if higher activity and expression could be achieved in a yeast host that usually does not hyperglycosylate. Using the alcohol oxidase promoter in conjunction with either the pho1 or the α-factor secretion signal, the recombinant enzyme was successfully secreted and glycosylated in P. pastoris. However, levels of protein expression from the chromosomally integrated vector were insufficient to detect activity.
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  • 59
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    Current genetics 29 (1996), S. 227-233 
    ISSN: 1432-0983
    Keywords: Trichoderma reesei ; β-Glucosidase ; Cellulase ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract An intronless form of thebgl1 gene encoding an extracellularβ-glucosidase fromTrichoderma reesei was expressed in the yeast Saccharomyces cerevisiae under the control of the yeast GAL 1 promoter. Transformation of a yeast strain with this vector resulted in transformants that produce and secrete activeβ-glucosidase into the growth medium. Additionally, active recombinantβ-glucosidase protein was shown to be localized predominantly in the periplasmic space by using ap-nitrophenylβ-D-glycoside hydrolysis assay against fractionated yeast cells. The apparent size of the recombinant enzyme was 10–15 kDa larger than that of the native form. Treatment of the recombinantβ-glucosidase with endoglycosidase-H indicated the apparent increase in size was due to N-linked glycosylation.
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  • 60
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    Current genetics 29 (1996), S. 227-233 
    ISSN: 1432-0983
    Keywords: Key words  Trichoderma reesei ; β-Glucosidase ; Cellulase ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract   An intronless form of the bgl1 gene encoding an extracellular β-glucosidase from Trichoderma reesei was expressed in the yeast Saccharomyces cerevisiae under the control of the yeast GAL1 promoter. Transformation of a yeast strain with this vector resulted in transformants that produce and secrete active β-glucosidase into the growth medium. Additionally, active recombinant β-glucosidase protein was shown to be localized predominantly in the periplasmic space by using a p-nitrophenyl β-D-glycoside hydrolysis assay against fractionated yeast cells. The apparent size of the recombinant enzyme was 10–15 kDa larger than that of the native form. Treatment of the recombinant β-glucosidase with endoglycosidase-H indicated the apparent increase in size was due to N-linked glycosylation.
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  • 61
    ISSN: 1432-0983
    Keywords: Key words Adenine biosynthesis ; ade8-18 ; ade2 mutations ; Red/white colony color assay ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In the yeast Saccharomyces cerevisiae the ade2, and/or the ade1, mutation in the adenine biosynthetic pathway leads to the accumulation of a cell-limited red pigment, while epistatic mutations in the same pathway, i.e. ade8, preclude this phenomenon, resulting in normal white colonies. The shift in color from red to white (or vice versa) with a combination of appropriate wild-type and mutant alleles of the adenine-pathway genes has been widely utilized as a non-selective phenotype to visualise and quantify the occurrence of various genetic events such as recombination, conversion and aneuploidy. It has provided an invaluable tool for the study of gene dosage and plasmid stability. In competition experiments between disrupted ade2, ade8-18 transformants carrying either a functional or non-functional episomal ADE8 gene, we verified that white ade8 ade2 cells show a remarkable selective advantage over red ade2 cells, with important implications on the use of this assay for the monitoring of genetic events. The accumulation of the red pigment in ade2 cells is likely to be the cause for impaired growth in these cells.
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  • 62
    ISSN: 1432-072X
    Keywords: Key words Plasma membrane H+-ATPase ; Saccharomyces cerevisiae ; Low pH ; PMA1 gene expression ; PMA2 gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cells of Saccharomyces cerevisiae grown in media with an initial pH of 2.5–6.0, acidified with a strong acid (HCl), exhibited the highest plasma membrane H+-ATPase-specific activity at an initial pH of 6.0. At a lower pH (above pH 2.5) ATPase activity (62–83% of the maximum level) still allowed optimal growth. At pH 2.5, ATPase activity was about 30% of the maximum value and growth was impaired. Quantitative immunoassays showed that the content of ATPase protein in the plasma membrane was similar across the entire pH range tested, although slightly lower at pH 2.5. The decrease of plasma membrane ATPase activity in cells grown at low pH was partially accounted for by its in vitro stability, which decreased sharply at pH below 5.5, although the reduction of activity was far below the values expected from in vitro measurements. Yeast growth under acid stress changed the pattern of gene expression observed at optimal pH. The level of mRNA from the essential plasma-membrane-ATPase-encoding gene PMA1 was reduced by 50% in cells grown at pH 2.5 as compared with cells grown at the optimal pH 5.0, although the content of ATPase in the plasma membrane was only modestly reduced. As observed in response to other kinds of stress, the PMA2 promoter at the optimal pH was up to eightfold more efficient in cells grown at pH 2.5, although it remained several hundred times less efficient than that of the PMA1 gene.
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  • 63
    ISSN: 1432-072X
    Keywords: Saccharomyces cerevisiae ; Pyruvate decarboxylase ; Pyruvate kinase ; Signalling ; Glycolysis mutants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Pyruvate decarboxylase, PDCase, activity in wild-type yeast cells growing on ethanol is quite low but increases up to tenfold upon addition of glucose, less with galactose and only slightly with glycerol. PDCase levels in glycolysis mutant strains growing on ethanol or acetate were higher than in the wild-type strain. These levels correlated with the sum of the concentrations of three-carbon glycolytic metabolites. The highest accumulation was observed in a fructose bisphosphate aldolase deletion mutant concomintant with the highest PDCase activity wild-type level. On the other hand, the PDCase levels in the different mutants again correlated with the sum of the concentrations of the three-carbon glycolytic metabolites. This was interpreted to mean that full induction of PDCase activity requires the accumulation of hexose-and triosephosphates.
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  • 64
    ISSN: 1432-072X
    Keywords: Saccharomyces cerevisiae ; Acetyl-CoA ; l-Lysine N6 ; acetytransferase ; Lysine catabolism
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    Topics: Biology
    Notes: Abstract The carbon catabolism of l-lysine starts in Saccharomyces cerevisiae with acetylation by an acetyl-CoA: l-lysine N6-acetyltransferase. The enzyme is strongly induced in cells grown on l-lysine as sole carbon source and has been purified about 530-fold. Its activity was specific for acetyl-CoA and, in addition to l-lysine, 5-hydroxylysine and thialysine act as acetyl acceptor. The following apparent Michaelis constants were determined: acetyl-CoA 0.8 mM, l-lysine 5.8 mM, dl-5-hydroxylysine 2.8 mM, l-thialysine 100 mM. The enzyme had a maximum activity at pH 8.5 and 37°C. Its molecular mass, estimated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis, was 52 kDa. Since the native molecular mass, determined by gel filtration, was 48 kDa, the enzyme is a monomer.
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  • 65
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    Journal of industrial microbiology and biotechnology 16 (1996), S. 102-109 
    ISSN: 1476-5535
    Keywords: Millet ; Pennisetum typhoides ; liquefaction ; saccharification ; baker's yeast ; Saccharomyces cerevisiae ; fermentation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract A fermentation medium based on millet (Pennisetum typhoides) flour hydrolysate and a four-phase feeding strategy for fed-batch production of baker's yeast,Saccharomyces cerevisiae, are presented. Millet flour was prepared by dry-milling and sieving of whole grain. A 25% (w/v) flour mash was liquefied with a thermostable 1,4-α-d-glucanohydrolase (EC 3.2.1.1) in the presence of 100 ppm Ca2+, at 80°C, pH 6.1–6.3, for 1 h. The liquefied mash was saccharified with 1,4-α-d-glucan glucohydrolase (EC 3.2.1.3) at 55°C, pH 5.5, for 2 h. An average of 75% of the flour was hydrolysed and about 82% of the hydrolysate was glucose. The feeding profile, which was based on a model with desired specific growth rate range of 0.18–0.23 h−1, biomass yield coefficient of 0.5 g g−1 and feed substrate concentration of 200 g L−1, was implemented manually using the millet flour hydrolysate in test experiments and glucose feed in control experiments. The fermentation off-gas was analyzed on-line by mass spectrometry for the calculation of carbon dioxide production rate, oxygen up-take rate and the respiratory quotient. Off-line determination of biomass, ethanol and glucose were done, respectively, by dry weight, gas chromatography and spectrophotometry. Cell mass concentrations of 49.9–51.9 g L−1 were achieved in all experiments within 27 h of which the last 15 h were in the fedbatch mode. The average biomass yields for the millet flour and glucose media were 0.48 and 0.49 g g−1, respectively. No significant differences were observed between the dough-leavening activities of the products of the test and the control media and a commercial preparation of instant active dry yeast. Millet flour hydrolysate was established to be a satisfactory low cost replacement for glucose in the production of baking quality yeast.
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  • 66
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    Molecular and cellular biochemistry 124 (1993), S. 131-140 
    ISSN: 1573-4919
    Keywords: Schizosaccharomyces pombe ; Saccharomyces cerevisiae ; H+-ATPase ; intracellular pH ; carboxy-seminaphthorhodafluor-1
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract We examined cytoplasmic pH regulation inSchizosaccharomyces pombe andSaccharomyces cerevisiae using pH-sensitive fluorescent dyes. Of several different fluorescent compounds tested, carboxy-seminaphthorhodafluor-1 (C.SNARF-1) was the most effective. Leakage of C.SNARF-1 fromS. pombe was much slower than leakage fromC. cerevisiae. Using the pH-dependent fluorescence of C.SNARF-1 we showed that at an external pH of 7, mean resting internal pH was 7.0 forS. pombe and 6.6 forS. cerevisiae. We found that internal pH inS. pombe was maintained over a much narrower range in response to changes in external pH, especially at acidic pH. The addition of external glucose caused an intracellular alkalinization in both species, although the effect was much greater inS. cerevisiae than inS. pombe. The plasma membrane H+-ATPase inhibitor diethylstilbestrol reduced both the rate and extent of alkalinisation, with an IC50 of approximately 35 μM in both species. Amiloride also inhibited internal alkalinisation with IC50's of 745 μM forS. cerevisiae and 490 μM forS. pombe.
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  • 67
    ISSN: 1573-1561
    Keywords: Encelia farinosa ; Asteraceae ; Trirhabda geminata ; Coleoptera ; Chrysomelidae ; feeding preferences ; acidic deposition ; plant stress
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Container grownEncelia farinosa were exposed to three 3-hr episodes of acidic fog (pH 2.5) typical of events in southern California. Adults and larvae of the specialist leaf-feeding herbivore,Trirhabda geminata, preferred to feed on the acidic-treated foliage compared to control fogged (pH 6.3–6.5) foliage. Previous feeding damage on the plants did not affect feeding preference. The acidic-fogged foliage was significantly higher in total nitrogen and soluble protein but not different from control-treated tissue in water content. Stress on native populations of this drought-deciduous shrub caused by atmospheric pollutants may also result in altered feeding ecology of the beetle.
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  • 68
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    Journal of chemical ecology 19 (1993), S. 395-410 
    ISSN: 1573-1561
    Keywords: Oreina gloriosa ; Coleoptera ; Chrysomelidae ; chemical defense ; cardenolides ; quantitative variation ; aging ; HPLC
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract The defensive secretion of the alpine chrysomelidOreina gloriosa is a complex mixture of mainly cardenolides and tyrosine betaine. Individually sampled secretions of adult laboratory-reared and field-collected beetles were analyzed by reverse-phase HPLC; 16 secretion components were quantified. Quantities and concentrations of different components were significantly affected by the age, sex, and reproductive status of individual beetles. Aging was correlated with marked increases (up to 4.4-fold) and decreases (up to 2.7-fold) of quantities and concentrations of several components. Differences between the sexes were smaller, but quantities of all components and concentrations of several components were larger in laboratory-reared females than in males. There was less of one component of the secretion in mated than unmated females, but the concentrations of four secretion components were higher (up to 1.6-fold) in mated females.
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  • 69
    ISSN: 1573-1561
    Keywords: Rapeseed ; Brassica spp. ; allyl isothiocyanate ; glucosinolates ; allelochemicals ; Limonius californicus (Mann.) ; Coleoptera ; Elateridae ; toxicity ; sublethal effects
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Acute toxicity of soil amended with allyl isothiocyanate (AITC) to three size classes ofLimonius californicus (Mann.) wireworms was determined in the laboratory. Wireworms were exposed to AITC at initial concentrations of 120–300 nmol/g soil for one day. During this time, extractable AITC concentrations decreased by 66 to 93 %. Probit analysis estimated LC50 values of 238 and 226 nmol/g soil at one day posttreatment for medium and large wireworms, respectively. For small wireworms, LC50 values decreased from 211 to 157 nmol/g soil during 1–137 days posttreatment. Sublethal concentrations of AITC significantly reduced feeding activity of treated wireworms at three posttreatment times and over the entire 137 days. Wireworm weight was not significantly affected by AITC. The potential exists to use glucosinolate-containing plant tissue as an isothiocyanate (ITC) source to reduce crop damage caused byL. californicus wireworms.
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  • 70
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    Journal of chemical ecology 19 (1993), S. 1219-1231 
    ISSN: 1573-1561
    Keywords: Host recognition behavior ; entomopathogenic nematodes ; feces ; Spodoptera exigua ; Lepidoptera ; Noctuidae ; Popillia japonica ; Coleoptera ; Scarabaeidae ; Blattella germanica ; Blatteria ; BlattellidaeAcheata domesticus ; Orthoptera ; Gryllidae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Host recognition by entomopathogenic nematodes may occur through contact with insects' excretory products, cuticle, or gut contents. We analyzed the behavioral responses of four species of entomopathogenic nematodes during contact with feces of natural or experimental hosts. Host recognition by nematodes was manifested in alterations in the frequency and/or duration of one or more search parameters including forward crawling, headwaving, body-waving, stopping, backward crawling, head-rubbing, and headthrusting.Heterorhabditis bacteriophora andSteinernema glaseri showed behavioral responses to contact with feces of their natural hosts,Spodoptera exigua (Lepidoptera) andPopillia japonica (Coleoptera), and to the experimental hosts,Acheata domesticus (Orthoptera) andBlatella germanica (Blatteria).Steinernema carpocapsae responded only toB. germanica feces, whereas5. scapterisci did not significantly respond to any of the insect species. During contact with cockroach feces, all nematodes, exceptS. scapterisci, showed avoidance behavior. We suggest that ammonia present in cockroach feces is inhibitory to nematodes. Specific host recognition by entomopathogenic nematodes may be an important mechanism to maintain host affinities.
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  • 71
    ISSN: 1573-1561
    Keywords: Holotrichia parallela ; large black chafer ; scarab beetle ; Coleoptera ; Scarabaeidae ; isoleucine methyl ester ; linalool ; sex pheromone ; circabidian periodicity ; pheromone titer
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract (R)-(−)-Linalool was identified as a minor component sex pheromone of the scarab beetleHolotrichia parallela (Coleoptera: Scarabaeidae). Field evaluations revealed that, although not attractive per se, (R)-(−)-linalool enhances the attractiveness of the major sex pheromone,L-isoleucine methyl ester (LIME). Analyses of the pheromone titers in the glands of field-collected females demonstrated the occurrence of peak levels of 48-hr (“circabidian”) periodicity. The levels of LIME in the glands of 45-day-old virgin females increased over three times from the scototo the photophase of a calling day, but the amounts of (R)-(−)-linalool did not significantly change. Virgin females had in average two times more LIME and 3.6 times more (R)-(−)-linalool than the average amount found in the field-captured beetles throughout the season.
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  • 72
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    Journal of chemical ecology 19 (1993), S. 1453-1459 
    ISSN: 1573-1561
    Keywords: Anomala schonfeldti ; Popillia japonica ; scarab beetle ; Coleoptera ; Scarabaeidae ; 2-(E)-nonenol ; sex pheromone ; mark-and-recapture ; field test ; mass trapping
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Synthetic 2-(E)-nonenol, previously identified as the sex pheromone ofAnomala schonfeldti (Coleoptera: Scarabaeidae), is demonstrated to be very attractive to males in the field. Nevertheless, no significant differences were found between treatments with 1, 5, 10, and 20 mg dosages. Males ofA. schonfeldti were more significantly attracted to traps at 30 cm high than at 90 cm. Although the observed behavior seemed to indicate a trend of more attraction to buried traps than those placed at 30 cm, there was no statistical difference between the two treatments. Pheromone-baited traps caught significantly more beetles than traps containing three virgin females. Over 70% of released beetles were recaptured in six traps surrounding the point of release and separated from each other by 50 m, suggesting a possible use of the pheromone (in combination with floral compounds) in mass trapping.
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  • 73
    ISSN: 1573-1561
    Keywords: Sitophilus granarius ; grain weevil ; Coleoptera ; aggregation pheromone ; electroantennogram ; coupled GC-EAG ; behavioral bioassay ; circular dichroism
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Coupled GC-EAG techniques have been applied to the study of volatiles from the grain weevil,Sitophilus granarius. for the first time. The size of EAG response was independent of the sex of the responding insect but was consistently larger to extracts of males than those of females. This difference was reflected in a behavioral preference for the male extracts by mated adults of both sexes tested together and virgin adults of both sexes tested separately. The GC-EAG results provide evidence for two materials that are released specifically by the males. Using circular dichroism. one has been found to be identical stereochemically with the (2S,3R)-sitophilate reported by others as the aggregation pheromone in a different strain. This enhances the prospects for the development of a single pheromone lure that would be generally applicable whatever the origin of the strain. The small amount of sitophilate found in the males suggests that it is not stored in large amounts. The other material, present in such a small amount that it has yet to be fully characterized, elicits a higher antennal activity than sitophilate and may have a significant role to play in enhancing the trap catch of this economically important pest.
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  • 74
    ISSN: 1573-1561
    Keywords: Coleoptera ; Staphylinidae ; Tenebrionidae ; larvae ; defensive glands ; quinone ; naphthoquinone ; taxonomy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract The abdominal glands of three bark-inhabiting larvae of generaBolitochara, Leptusa (Staphylinidae), andHypophloeus (Tenebrionidae) were studied chemically and morphologically. Behavior of the larvae indicated that secretion is emitted only after severe disturbance of the larvae. These mechanical contacts may also occur incidentally with coinhabiting nonpredatory arthropods when the beetle larvae move within small interstices under bark. Depending on the species, the secretions contained 1,4-benzoquinone derivatives and three 6-alkyl-naphthoquinones dissolved within various alkanes, alkenes, ethyl-, isopropyl-, and isoamylesters. More erratically distributed gland constituents also detected were acetophenone, benzyl propionate, and methyl hydroxybenzoate. In the laboratory, synthetic quinone-containing solutions simulating those found inLeptusa andBolitochara larvae acted as strong topical irritants and caused further damage to last-stageCalliphora vomitoria larvae if hydrocarbons or esters were used as solvents. The natural secretions ofHypophloeus versipellis elicited considerable mortality in two subcortical sciarid larvae cooccurring with tenebrionid larvae. Bioassay and secretion chemistry of the Staphylinidae/Tenebrionidae larval secretions indicated that they are typical defensive secretions that act topically. Morphological data characterized Bolitocharini larvae as possessing protuberant abdominal tergites supplied with an interiorly situated gland reservoir. After mechanical contact, the defensive secretion is topically applied to other arthropods by dabbing this tergal protuberance on targets. The defensive gland ofHypophloeus versipellis is unusual in possessing a movable reservoir opening situated at the anterior border of tergite IX. By this peculiar gland morphologyHypophloeus larvae are capable of shooting secretion droplets frontally from their slightly depressed dorsal abdominal surface without bending their abdominal tips dorsally. This seems an adaptation to the interstitial habitat of the larvae. The types of defensive glands and their phylogenetic value in Aleocharinae/Tenebrionidae larvae are discussed.
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  • 75
    ISSN: 1573-1561
    Keywords: Hylastes cunicularius ; Hylastes brunneus ; Hylastes opacus ; Dryocoetes autographus ; Hylobius abietis ; Hylobius pinastri ; α-pinene ; terpenes ; turpentine ; ethanol ; ground traps ; Coleoptera ; Scolytidae ; Curculionidae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Responses of threeHylastes species,Dryocoetes autographus, and twoHylobius species to terpenes and ethanol were studied in field experiments on clear-cut forest sites in Sweden using baited ground traps.α-Pinene alone did not attract any of the six species. A terpene blend (spruce turpentine consisting mainly ofα-pinene,β-pinene, and 3-carene) attractedHylastes cunicularius, H. brunneus, andHylobius abietis in some experiments, but not in others. The attractiveness of ethanol also varied; the only species consistently attracted wasH. abietis. Baits containing both terpenes and ethanol, particularly the combination of spruce turpentine and ethanol, were attractive to all species exceptHylobius pinastri. InH. abietis, the terpene plus ethanol/ ethanol catch ratios increased during early summer. Seasonal differences in catch levels were observed inH. cunicularius andH. abietis. The addition ofα-pinene reduced the attractiveness of the combination of spruce turpentine and ethanol toH. cunicularius, H. opacus, andD. autographus. The differences in response to the volatiles between species are probably related to differences in reproductive behavior and host preferences.
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  • 76
    ISSN: 1573-4919
    Keywords: osmotic stress ; Saccharomyces cerevisiae ; glycerol ; K+/Na+ ions ; osmoregulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The intracellular level of Na+ and K+ of S. cerevisiae strain AB1375 revealed that under KCl as well as sorbitol stress, the cationic level was comparable to the level under no stress conditions. On the other hand, there was a sharp drop in the intracellular K+ content and increase in the Na+ content on addition of NaCl to the medium. However, the total cationic level was close to that under control conditions. In addition to changes in the cationic level, an enhanced production and accumulation of glycerol were also observed under osmotic stress. A regulatory mechanism co-ordinating the intracellular concentration of glycerol as well as Na+, K+ content under osmotic stress conditions has been proposed.
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  • 77
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    Plant molecular biology 22 (1993), S. 1177-1180 
    ISSN: 1573-5028
    Keywords: abscisic acid ; developmental regulation ; heat shock proteins ; Oryza sativa ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Antibodies raised against yeast heat shock protein (HSP) 104 recognized a heat-inducible polypeptide with a molecular mass of 110 kDa in shoot tissue of young rice seedlings. Root tissue of the same age showed no immuno-reaction with yeast HSP 104 antibodies. The 110 kDa polypeptide of rice was also shown to be abscisic acid-inducible in young seedlings. Though this polypeptide was seen to be constitutively present in the flag leaf of 90-day-old field-grown plant, it was not much affected by either heat shock or abscisic acid in this case.
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  • 78
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    Journal of fluorescence 3 (1993), S. 241-244 
    ISSN: 1573-4994
    Keywords: Killer toxin K1 ; bromocresol purple staining ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Physics
    Notes: Abstract Optimal assay conditions were established for the previously described method used to determine the activity ofSaccharomyces cerevisiae pore-forming killer toxin K1. The method is based on cell staining with bromocresol purple. Sensitive cells ofS. cerevisiae from the early exponential phase under nongrowth conditions and in the presence of glucose were the most convenient for determining the killer toxin activity. Maximum killing war reached when the suspension was buffered with 10 mM citrate-phosphate at pH 4.6.
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  • 79
    ISSN: 1573-5028
    Keywords: MAP kinase ; osmotic stress ; Pisum sativum ; Saccharomyces cerevisiae ; signal transduction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Previous analysis of the MAP kinase homologue from Pisum sativum (PsMAPK) revealed a potential MAP kinase motif homologous to that found in eukaryotic cdc2 kinases. Sequence comparison showed a 47% identity on amino acid sequence basis to the Saccharomyces cerevisiae Hog 1p MAP kinase involved in the osmoregulatory pathway. Under conditions of salt-stress aberrant morphology of a hog1 deletion mutant was completely restored and growth was partially restored by expression of the PsMAPK. This shows that PsMAPK is functionally active as a MAP kinase in S. cerevisiae. Comparison of PsMAPK with other kinases involved in osmosensitivity, showed a high degree of homology and implicates a possible role for PsMAPK in a P. sativum osmosensing signal transduction pathway.
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  • 80
    ISSN: 1617-4623
    Keywords: Key words Saccharomyces douglasii ; Saccharomyces cerevisiae ; CBP2 ; Mitochondria ; Pre-mRNA processing
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  In Saccharomyces cerevisiae the only known role of the CBP2 gene is the excision of the fifth intron of the mitochondrial cyt b gene (bI5). We have cloned the CBP2 gene from Saccharomyces douglasii (a close relative of S. cerevisiae). A comparison of the S. douglasii and S. cerevisiae sequences shows that there are 14% nucleotide substitutions in the coding region, with transitions being three times more frequent than transversions. At the protein level sequence identity is 87%. We have demonstrated that the S. douglasii CBP2 gene is essential for respiratory growth in the presence of a wild-type S. douglasii mitochondrial genome, but not in the presence of an intronless S. cerevisiae mitochondrial genome. Also the S. douglasii and S. cerevisiae CBP2 genes are completely interchangeable, even though the intron bI5 is absent from the S. douglasii mitochondrial genome.
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  • 81
    ISSN: 1617-4623
    Keywords: Pre-mRNA splicing ; Saccharomyces cerevisiae ; Suppressors ; prp24-1
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The temperature-sensitiveprp24-1 mutation defines a gene product required for the first step in pre-mRNA splicing. PRP24 is probably a component of the U6 snRNP particle. We have applied genetic reversion analysis to identify proteins that interact with PRP24. Spontaneous revertants of the temperaturesensitive (ts)prp24-1 phenotype were analyzed for those that are due to extragenic suppression. We then extended our analysis to screen for suppressors that confer a distinct conditional phenotype. We have identified a temperature-sensitive extragenic suppressor, which was shown by genetic complementation analysis to be allelic toprp21-1. This suppressor,prp21-2, accumulates pre-mRNA at the non-permissive temperature, a phenotype similar to that ofprp21-1. prp21-2 completely suppresses the splicing defect and restores in vivo levels of the U6 snRNA in theprp24-1 strain. Genetic analysis of the suppressor showed thatprp21-2 is not a bypass suppressor ofprp24-1. The suppression ofprp24-1 byprp21-2 is gene specific and also allele specific with respect to both the loci. Genetic interactions with other components of the pre-spliceosome have also been studied. Our results indicate an interaction between PRP21, a component of the U2 snRNP, and PRP24, a component of the U6 snRNP. These results substantiate other data showing U2–U6 snRNA interactions.
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  • 82
    ISSN: 1617-4623
    Keywords: Saccharomyces douglasii ; Saccharomyces cerevisiae ; CBP2 ; Mitochondria ; Pre-mRNA processing
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract InSaccharomyces cerevisiae the only known role of theCBP2 gene is the excision of the fifth intron of the mitochondrialcyt b gene (bI5). We have cloned theCBP2 gene fromSaccharomyces douglasii (a close relative ofS. cerevisiae). A comparison of theS. douglasii andS. cerevisiae sequences shows that there are 14% nucleotide substitutions in the coding region, with transitions being three times more frequent than transversions. At the protein level sequence identity is 87%. We have demonstrated that theS. douglasii CBP2 gene is essential for respiratory growth in the presence of a wild-typeS. douglasii mitochondrial genome, but not in the presence of an intronlessS. cerevisiae mitochondrial genome. Also theS. douglasii andS. cerevisiae CBP2 genes are completely interchangeable, even though the intron bI5 is absent from theS. douglasii mitochondrial genome.
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  • 83
    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; Nucleotide excision repair ; RAD14 ; XPA homologue
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In the accompanying paper we demonstrated that endonuclease III-sensitive sites in theMATα andHMLα loci ofSaccharomyces cerevisiae are repaired by the Nucleotide Excision Repair (NER) pathway. In the current report we investigated the repair of endonuclease III sites, 6-4 photoproducts and cyclobutane pyrimidine dimers (CPDs) in arad14-2 point mutant and in arad14 deletion mutant. TheRAD14 gene is the yeast homologue of the human gene that complements the defect in cells from xeroderma pigmentosum (XP) patients belonging to complementation group A. In the point mutant we observed normal repair of endonuclease III sites (i.e. as wild type), but no removal of CPDs at theMATα andHMLα loci. Similar experiments were undertaken using the recently createdrad14 deletion mutant. Here, neither endonuclease III sites nor CPDs were repaired inMAT a orHMR a. Thus the point mutant appears to produce a gene product that permits the repair of endonuclease III sites, but prevents the repair of CPDs. Previously it was found that, in the genome overall, repair of 6-4 photoproducts was less impaired than repair of CPDs in the point mutant. The deletion mutant repairs neither CPDs nor 6-4 photoproducts in the genome overall. This finding is consistent with the RAD14 protein being involved in lesion recognition in yeast. A logical interpretation is that therad14-2 point mutant produces a modified protein that enables the cell to repair endonuclease III sites and 6-4 photoproducts much more efficiently than CPDs. This modified protein may aid studies designed to elucidate the role of the RAD14 protein in lesion recognition.
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  • 84
    ISSN: 1617-4623
    Keywords: Key words RCC1 ; Saccharomyces cerevisiae ; Serine/threonine protein kinase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  The yeast PRP20 protein is homologous to the RCC1 protein of higher eukaryotes and is required for mRNA export and maintenance of nuclear structure. RCC1/PRP20 act as guanine nucleotide exchange factors for the nuclear Ras-like Ran/GSP1 proteins. In a search for prp20-10 allele-specific high-copy-number suppressors, the KSP1 locus, encoding a serine/threonine protein kinase was isolated. Ksp1p is a nuclear protein that is not essential for vegetative growth of yeast. Inactivation of the kinase activity by a mutation affecting the catalytic center of the Ksp1p eliminated the suppressing activity. Based on the isolation of a protein kinase as a high-copy-number suppressor, the phosphorylation of Prp20p was examined. In vivo labeling experiments showed that Prp20p is a phosphoprotein; however, deletion of the KSP1 kinase did not affect Prp20p phosphorylation.
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  • 85
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    Molecular genetics and genomics 253 (1996), S. 111-117 
    ISSN: 1617-4623
    Keywords: Key words MEL gene ; α-galactosidase ; Saccharomyces cerevisiae ; Saccharomyces paradoxus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  In order to study the molecular evolution of the yeasts grouped in the Saccharomyces sensu stricto species complex by analysis of the MEL gene family, we have cloned and sequenced two new species-specific MEL genes from Saccharomyces yeasts: S. paradoxus (MELp) and a Japanese Saccharomyces sp. (MELj). The clones were identified by sequence homology to the S. cerevisiae MEL1 gene. Both clones revealed an ORF of 1413 bp coding for a protein of 471 amino acids. The deduced molecular weights of the α-galactosidase enzymes were 52 767 for MELp and 52 378 for MELj. The nucleotide sequences of the MELp (EMBL accession no. X95505) and the MELj (EMBL accession no. X95506) genes showed 74.7% identity. The degree of identity of MELp to the MEL1 gene was 76.8% and to the S. pastorianus MELx gene, 75.7%. The MELj coding sequence was 75.1% identical to the MEL1 gene and 80.7% to the MELx gene. The data suggest that MEL1, MELj, MELp, and MELx genes are species-specific MEL genes. The strains studied each have only one MEL locus. The MELp gene is located on the S. paradoxus equivalent of S. cerevisiae chromosome X; the MELj gene was on the chromosome that comigrates with the S. cerevisiae chromosome VII/XV doublet and hybridizes to the S. cerevisiae chromosome XV marker HIS3.
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  • 86
    ISSN: 1617-4623
    Keywords: Key words DEAD-box protein ; DED1 ; RCC1 ; Saccharomyces cerevisiae ; SRM1
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  The Saccharomyces cerevisiae temperature-sensitive mutants srm1-1, mtr1-2 and prp20-1 carry alleles of a gene encoding a homolog of mammalian RCC1. In order to identify a protein interacting with RCC1, a series of suppressors of the srm1-1 mutation were isolated as cold-sensitive mutants and one of the mutants, designated ded1-21, was found to be defective in the DED1 gene. The double mutant, srm1-1 ded1-21, could grow at 35° C, but not at 37° C. A revertant of srm1-1 ded1-21 that became able to grow at 37° C acquired another mutation in the SRM1 gene, indicating the tight relationship between SRM1 and DED1. In all the rcc1 - strains examined, the amount of mutated SRM1 proteins was reduced or not detectable at the nonpermissive temperature. While mutated SRM1 protein was stabilized in all of the rcc1 - strains by the ded1-21 mutation, the ded1-21 mutation suppressed both srm1-1 and mtr1-2, but not the prp20-1 mutation, contrary to the previous finding that overproduction of the S. cerevisiae Ran homolog GSP1 suppresses prp20-1, but not srm1-1 or mtr1-2.
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  • 87
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    Molecular genetics and genomics 251 (1996), S. 682-691 
    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; Two-hybrid system ; Protein-protein interactions ; PKC1 pathway ; MAP kinase cascade
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The two-hybrid system for the identification of protein-protein interactions was used to screen for proteins that interact in vivo with theSaccharomyces cerevisiae Pkc1 protein, a homolog of mammalian protein kinase C. Four positive clones were isolated that encoded portions of the protein kinase Mkk1, which acts downstream of Pkc1p in thePKC1-mediated signalling pathway. Subsequently, Pkc1p and the otherPKC1 pathway components encoding members of a MAP kinase cascade, Bck1p (a MEKK), Mkk1p, Mkk2p (two functionally homologous MEKs), and Mpk1p (a MAP kinase), were tested pairwise for interaction in the two-hybrid assay. Pkc1p interacted specifically with small N-terminal deletions of Mkk1p, and no interaction between Pkc1p and any of the other known pathway components could be detected. Interaction between Pkc1p and Mkk1p, however, was found to be independent of Mkk1p kinase activity. Bck1p was also found to interact with Mkk1p and Mkk2p, and the interaction required only the predicted C-terminal catalytic domain of Mkk1p. Furthermore, we detected protein-protein interactions between two Bck1p molecules via their N-terminal regions. Finally, Mkk2p and Mpk1p also interacted in the two-hybrid assay. These results suggest that the members of thePKC1-mediated MAP kinase cascade form a complex in vivo and that Pkc1p is capable of directly interacting with at least one component of this pathway.
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  • 88
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    Molecular genetics and genomics 251 (1996), S. 707-715 
    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; Flocculation ; Transcriptional regulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A flocculation gene was cloned from aSaccharomyces cerevisiae ATCC60715 genomic library, known to contain theFLO8 gene, on the basis of its ability to confer a flocculation phenotype on a non-flocculent strain. From a total of 11 130 clones, four clones sharing the several restriction fragments were isolated, suggesting that these were derived from the same locus. The results of integration mapping and disruption of the cloned gene indicated that this gene was theFLO8 gene. After disruption of theFLO8 gene, the strain lost its ability to flocculate. The DNA sequence of theFLO8 gene was determined. This gene includes a 2187-bp open reading frame that encodes a 729-amino acid protein. Computer analysis indicated that theFLO8 gene has a significant degree of homology with aS. cerevisiae chromosome V DNA sequence, but no homology with theFLO1 gene. The hydrophobicity profile of the putativeFLO8 gene product did not indicate the presence of any significantly hydrophobic regions. Southern analysis of theFLO8 gene present in various yeast strains indicated that theFLO8 gene is highly conserved in yeast strains having a variety of flocculation phenotypes and genotypes. Northern analysis revealed that the level ofFLO1 gene transcription is dependent on the rate of transcription of theFLO8 gene. These results suggest that theFLO8 gene mediates flocculation via transcriptional activation of theFLO1 gene.
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  • 89
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    Molecular genetics and genomics 250 (1996), S. 395-404 
    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; DNase ; Eukaryotes ; Genetic recombination
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Site-specific endonucleases have been found in various eukaryotic organelles such as mitochondria, chloroplasts and nuclei. These endonucleases initiate site-specific or homologous gene conversion in mitochondrial and nuclear DNA. Here, we report a new site-specific endonuclease activity, Endo.SK1, identified in mitochondria of strain SK1, a homothallic diploid strain ofSaccharomyces cerevisiae. Nucleotide sequences around the Endo.SK1-cleavage sites are different from those of known yeast site-specific endonucleases. The Endo.SK1 activity is, at least partly, specified by a gene in the SK1-derived mitochondria. A novel feature of the Endo.SK1 activity is its inducibility: the endonuclease activity was induced by ca. 40-fold by transfer of cells from a glucose medium into an acetate medium, and was then repressed. This transient induction was independent of the ploidy level of the cells, and coincided with induction of fumarase, a mitochondrial enzyme involved in the TCA cycle. Co-induction and co-repression of the mitochondrial site-specific endonuclease activity and a respiration-related enzyme indicate that the endonuclease activity is regulated in response to physiological conditions, and suggest a possible role for the endonuclease in mitochondrial DNA metabolism.
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  • 90
    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; Calcineurin ; MAP kinase cascade ; Pheromone-induced growth arrest ; Synthetic effect
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Saccharomyces cerevisiae mutants which exhibit phenotypes (calcium resistance and vanadate sensitivity) similar to those of calcineurin-deficient mutants were isolated. The mutants were classified into four complementation groups (crv1,2,3 and4).crv1 was allelic tocnb1, a mutation in the regulatory subunit of calcineurin. The nucleotide sequences ofCRV2 andCRV3 genes which complemented thecrv2 andcrv3 mutations, respectively, are identical to those ofBCK1/SLK1/SKC1/SSP31 andMPK1/SLT2, respectively, which are both involved in the MAP kinase cascade. A calcineurin-deletion mutation (Δcnb1), which by itself has no detectable effect on growth and morphology, enhanced some phenotypes (slow growth and morphological abnormality) ofcrv2 andcrv3 mutants. These phenotypes ofcrv2 andcrv3 mutants were partially suppressed by Ca2+ or by overproduction of the calcineurin subunits (Cmp2 and Cnb1). Like the calcineurin-deficient mutant,crv2 andcrv3 mutants were defective in recovery from α-factor-induced growth arrest. The defect in recovery of the Δcnb1 mutant was suppressed by overexpression ofMPK1. These results indicated that the calcineurin-mediated and the Mpk1- (Bck1-) mediated signaling pathways act in parallel to regulate functionally redundant cellular events important for growth.
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  • 91
    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; Ribosomal protein genes ; Transcription activation ; cAMP ; Growth control
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The rate of ribosomal protein gene (rp-gene) transcription in yeast is accurately adjusted to the cellular requirement for ribosomes under various growth conditions. However, the molecular mechanisms underlying this co-ordinated transcriptional control have not yet been elucidated. Transcriptional activation of rp-genes is mediated through two different multifunctional trans-acting factors, ABF1 and RAP1. In this report, we demonstrate that changes in cellular rp-mRNA levels during varying growth conditions are not parallelled by changes in the in vitro binding capacity of ABF1 or RAP1 for their cognate sequences. In addition, the nutritional upshift response of rp-genes observed after addition of glucose to a culture growing on a non-fermentative carbon source turns out not to be the result of increased expression of the ABF1 and RAP1 genes or of elevated DNA-binding activity of these factors. Therefore, growth rate-dependent transcription regulation of rp-genes is most probably not mediated by changes in the efficiency of binding of ABF1 and RAP1 to the upstream activation sites of these genes, but rather through other alterations in the efficiency of transcription activation. Furthermore, we tested the possibility that cAMP may play a role in elevating rp-gene expression during a nutritional shift-up. We found that the nutritional upshift response occurs normally in several mutants defective in cAMP metabolism.
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  • 92
    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; UV damage ; Mating type ; Inducible repair
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The prior UV irradiation of α haploid Saccharomyces cerevisiae with a UV dose of 25 J/m2 substantially increases the repairability of damage subsequently induced by a UV dose of 70 J/m2 given 1 h after the first irradiation. This enhancement of repair is seen at both the MATa and HMLα loci, which are, respectively, transcriptionally active and inactive in α haploid cells. The presence in the medium of the protein synthesis inhibitor, cycloheximide in the period between the two irradiations eliminated this effect. Enhanced repair still occurred if cycloheximide was present only after the final UV irradiation. This indicated that the first result is not due to cycloheximide merely blocking the synthesis of repair enzymes associated with a hypothetical rapid turnover of such molecules. The enhanced repairability is not the result of changes in chromatin accessibility without protein synthesis, merely caused by the repair of the damage induced by the prior irradiation. The data clearly show that a UV-inducible removal of pyrimidine dimers has occurred which involves the synthesis of new proteins. The genes known to possess inducible promoters, and which are involved in excision are RAD2, RAD7, RAD16 and RAD23. Studies with the rad7 and rad16 mutants which are defective in the ability to repair HMLα and proficient in the repair' of MATα showed that in rad7, preirradiation enhanced the repair at MATα, whereas in rad16 this increased repair of MATα was absent. The preirradiation did not modify the inability to repair HMLα in either strain. Thus RAD16 has a role in this inducible repair. Inducible repair is also absent in a rad2 strain which cannot repair MATα or HMLα after a single UV dose.
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  • 93
    ISSN: 1572-9710
    Keywords: saproxylic ; Coleoptera ; spruce forest ; spatial scale ; conservation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: The diversity in different groups of obligate saproxylic beetles was related to ecological variables at three levels of spatial scale in mature spruce-dominated forest. The variables were connected to: (i) decaying wood, (ii) wood-inhabiting fungi, (iii) the level of disturbance, (iv) landscape ecology, and (v) vegetational structure. Several strong relationships were found at medium (1 km2) and large scales (4 km2), while only weak relationships were found at a small scale (0.16 ha; 1 ha=104 m2). This may be explained by the local variations in habitat parameters and the high mobilities of many beetle species. Factors connected to decaying wood and wood-inhabiting fungi were clearly the most important factors at all scale levels. In particular, the variables diversity of dead tree parts, number of dead trees of large diameter and number of polypore fungi species increased the species richness of many groups and increased the abundance of many species. Eight species were absent below a certain density of decaying wood per 1 or 4 km2. Former extensive cutting was a negative factor at large scale, probably because of decreasing recolonization with increasing distance to the source habitats. Thinning reduced the diversity of species associated with birch. The development of guidelines favouring the diversity of saproxylic beetles are discussed below.
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  • 94
    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; Sodium efflux ; Lithium efflux ; ATPase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The ENA2 gene encoding a P-type ATPase involved in Na+ and Li+ effluxes in Saccharomyces cerevisiae has been isolated. The putative protein encoded by ENA2 differs only in thirteen amino acids from the protein encoded by ENA1/PMR2. However, ENA2 has a very low level of expression and for this reason did not confer significant Li+ tolerance on a Li+ sensitive strain. ENA1 and ENA2 are the first two units of a tandem array of four highly homologous genes with probably homologous functions.
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  • 95
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    Molecular genetics and genomics 237 (1993), S. 375-384 
    ISSN: 1617-4623
    Keywords: Regulation of meiosis ; Saccharomyces cerevisiae ; IME1
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The IME1 gene of Saccharomyces cerevisiae is required for initiation of meiosis. Transcription of IME1 is detected under conditions which are known to induce initiation of meiosis, namely starvation for nitrogen and glucose, and the presence of MATa1 and MATα2 gene products. In this paper we show that IME1 is also subject to translational regulation. Translation of IME1 mRNA is achieved either upon nitrogen starvation, or upon G1 arrest. In the presence of nutrients, constitutively elevated transcription of IME1 is also sufficient for the translation of IME1 RNA. Four different conditions were found to cause expression of Imel protein in vegetative cultures: elevated transcription levels due to the presence of IME1 on a multicopy plasmid; elevated transcription provided by a Gal-IME1 construct; G1 arrest due to α-factor treatment; G1 arrest following mild heat-shock treatment of cdc28 diploids. Using these conditions, we obtained evidence that starvation is required not only for transcription and efficient translation of IME1, but also for either the activation of Ime1 protein or for the induction/activation of another factor that, either alone or in combination with Ime1, induces meiosis.
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  • 96
    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; Transcription ; spt mutants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Mutations in the SPT4 gene of Saccharomyces cerevisiae were isolated as suppressors of δ insertion mutations that interfere with adjacent gene transcription. Recent genetic evidence indicates that the SPT4 protein functions with two other proteins, SPT5 and SPT6, in some aspect of transcription initiation. In this work we have characterized the SPT4 gene and we demonstrate that spt4 mutations, like spt5 and spt6 mutations, cause changes in transcription. Using the cloned SPT4 gene, spt4 null mutations were constructed; in contrast to spt5 and spt6 null mutants, which are inviable, spt4 null mutants are viable and have an Spt− phenotype. The DNA sequence of the SPT4 gene predicts a protein product of 102 amino acids that contains four cysteine residues positioned similarly to those of zinc binding proteins. Mutational analysis suggests that at least some of these cysteines are essential for SPT4 function. Genetic mapping showed that SPT4 is a previously unidentified gene that maps to chromosome VII, between ADE6 and CLY8.
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  • 97
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    Molecular genetics and genomics 237 (1993), S. 463-466 
    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; cyrl-2 ; Nonsense mutation ; CAMP
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary cyrl-2 is a temperature-sensitive mutation of the yeast adenylate cyclase structural gene, CYR1. The cyrl-2 mutation has been suggested to be a UGA mutation since a UGA suppressor SUP201 has been isolated as a suppressor of the cyrl-2 mutation. Construction of chimeric genes restricted the region containing the cyrl-2 mutation, and the cyrl-2 UGA mutation was identified at codon 1282, which lies upstream of the region coding for the catalytic domain of adenylate cyclase. Alterations in the region upstream of the cyrl-2 mutation site result in null mutations. The complete open reading frame of the cyrl-2 gene expressed under the control of the GAL1 promoter complemented cyrl-dl in a galactose-dependent manner. These results suggest that at the permissive temperature weak readthrough occurs at the cyrl-2 mutation site to produce low levels of active adenylate cyclase. An endogenous suppressor in yeast cells is assumed to be responsible for this readthrough.
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  • 98
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    Molecular genetics and genomics 238 (1993), S. 6-16 
    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; cAMP MKS1 ; GAL11
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In order to isolate genes that function downstream of the Ras-cAMP pathway in Saccharomyces cerevisiae, a YEp13-based genomic library was screened for clones that inhibit growth of cells with diminished A-kinase activity. One such gene, MKS1, was found to encode a hydrophilic 52 kDa protein that shares weak homology with the yeast SPT2/SIN1 gene product. Three lines of evidence suggest that the MKS1 gene product is a negative regulator downstream of the Ras-cAMP pathway: (i) overexpression of MKS1 inhibits growth of cyrl disruptant cells on YPD medium containing a low concentration of cAMP; (ii) overexpression of MKS1 does not affect TPK1 expression; and (iii) the temperature-sensitive cyrl-230 mutation is partially suppressed by mks1 disruption. The mks1 mutant shows similar phenotypes to gal11/spt13, i.e., it cannot grow on YPGal containing ethidium bromide at 25°C, or on YPGly or SGal at 37°C. The mks1 gal11 double mutant shows more marked phenotypic changes than the single mutants. These results suggest that MKS1 is involved in transcriptional regulation of several genes by cAMP.
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  • 99
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    Molecular genetics and genomics 240 (1993), S. 36-42 
    ISSN: 1617-4623
    Keywords: Yeast ; Saccharomyces cerevisiae ; DNA synthesis genes ; Cell cycle regulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Two mutants have been isolated in Saccharomyces cerevisiae in which transcripts from at least CDC8, CDC9, CDC21 (TMP1) and POL1 genes are expressed constitutively in cells blocked at START by use of either α-pheromone or the cdc28 mutation. The transcripts from these genes also persist in mutant stationary phase cells; however, cell cycle regulation of these four DNA synthesis genes occurs normally in late G1. The mutation therefore does not appear to lie in the MCB-DSC1 (MBF) system that controls the periodic regulation of the genes, but must affect some control mechanism regulating basal levels of expression.
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  • 100
    ISSN: 1617-4623
    Keywords: Heat shock response ; HSP70 ; Saccharomyces cerevisiae ; RAS-CAMP pathway ; Multicopy suppressor of ira1
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: abstract The MSI3 gene was isolated as a multicopy suppressor of the heat shock-sensitive phenotype of the iral mutation, which causes hyperactivation of the RAS-cAMP pathway. Overexpression of MSI3 also suppresses the heat shock-sensitive phenotype of the bcyl mutant. Determination of the DNA sequence of MSI3 revealed that MSI3 can encode a 77.4 kDa protein related to the HSP70 family. The amino acid sequence of Msi3p is about 30% identical to that of the Ssalp of Saccharomyces cerevisiae. This contrasts with the finding that members of the HSP70 family generally show at least 50% amino acid identity. The consensus nucleotide sequence of the heat shock element (HSE) was found in the upstream region of MSI3. Moreover, the steady-state levels of the MSI3 mRNA and protein were increased upon heat shock. These results indicate that the MSI3 gene encodes a novel HSP70-like heat shock protein. Disruption of the MSI3 gene was associated with a temperature sensitive growth phenotype but unexpectedly, thermotolerance was enhanced in the disruptant.
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