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  • Saccharomyces cerevisiae  (86)
  • nitrogen  (82)
  • Springer  (168)
  • American Association of Petroleum Geologists (AAPG)
  • American Meteorological Society
  • 1995-1999
  • 1985-1989  (168)
  • 1970-1974
  • 1989  (86)
  • 1988  (82)
  • 1971
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  • 1995-1999
  • 1985-1989  (168)
  • 1970-1974
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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Journal of industrial microbiology and biotechnology 4 (1989), S. 49-53 
    ISSN: 1476-5535
    Keywords: l-Phenylacetyl carbinol ; Saccharomyces cerevisiae ; Yeast ; Benzaldehyde ; Biotransformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary The rate of production ofl-phenylacetyl carbinol bySaccharomyces cerevisiae in reaction mixtures containing benzaldehyde with sucrose or pyruvate as cosubstrate was investigated in short 1 h incubations. The effect of yeast dose rate, sucrose and benzaldehyde concentration and pH on the rate of reaction was determined. Maximum biotransformation rates were obtained with concentrations of benzaldehyde, sucrose and yeast of 6 g, 40 g and 60 g/l, respectively. Negligible biotransformation rates were observed at a concentration of 8 g/l benzaldehyde. The reaction had a pH optimum of 4.0–4.5. Rates of bioconversion of benzaldehyde and selected substituted aromatic aldehydes using both sucrose and sodium pyruvate as cosubstrate were compared. The rate of aromatic alcohol production was much higher when sucrose was used rather than pyruvate.o-Tolualdehyde and 1-chlorobenzaldehyde were poor substrates for aromatic carbinol formation although the latter produced significant aromatic alcohol in sucrose-containing media. Yields of 2.74 and 3.80 g/l phenylacetyl carbinol were produced from sucrose and pyruvate, respectively, in a 1 h reaction period.
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  • 2
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    Journal of industrial microbiology and biotechnology 4 (1989), S. 81-84 
    ISSN: 1476-5535
    Keywords: Ethanol fermentation ; Wheat starch ; Saccharomyces cerevisiae ; immobilization ; Continuous dynamic immobilized biocatalyst bioreactor ; Biocatalyst bioreactor
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary A simple and efficient method of conversion of wheat starch B to ethanol was investigated. Employing a two-stage enzymatic saccharification process, 95% of the wheat starch was converted to fermentable sugars in 40 h. From 140 g/l total sugars in the feed solution, 63.6 g/l ethanol was produced continuously with a residence time of 3.3 h in a continuous dynamic immobilized biocatalyst bioreactor by immobilized cells ofSaccharomyces cerevisiae. The advantages and the application of this bioreactor to continuous alcoholic fermentation of industrial substrates are presented.
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  • 3
    ISSN: 1572-8773
    Keywords: Manganese ; Electron spin resonance ; Superoxide dismutase ; Saccharomyces cerevisiae ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Summary Manganese accumulation was studied by room-temperature electron spin resonance (ESR) spectroscopy inSaccharomyces cerevisiae grown in the presence of increasing amounts of MnSO4. Mn2+ retention was nearly linear in intact cells for fractions related to both low-molecular-mass and macromolecular complexes (‘free’ and ‘bound’ Mn2+, respectively). A deviation from linearity was observed in cell extracts between the control value and 0.1 mM Mn2+, indicating more efficient accumulation at low Mn2+ concentrations. The difference in slopes between the two straight lines describing Mn2+ retention at concentrations lower and higher than 0.1 mM, respectively, was quite large for the free Mn2+ fraction. Furthermore it was unaffected by subsequent dialyses of the extracts, showing stable retention in the form of low-molecular-mass complexes. In contrast, the slope of the line describing retention of ‘bound’ Mn2+ at concentrations higher than 0.1 mM became less steep after subsequent dialyses of the cell extracts. This result indicates that the macromolecule-bound Mn2+ was essentially associated with particulate structures. In contrast to Cu2+, Mn2+ had no effect on the major enzyme activities involved in oxygen metabolism except for a slight increase of cyanide-resistant Mn-superoxide dismutase activity, due to dialyzable Mn2+ complexes.
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  • 4
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    Springer
    Entomologia experimentalis et applicata 51 (1989), S. 175-187 
    ISSN: 1570-7458
    Keywords: Anticarsia ; compensatory feeding behavior ; diet dilution ; food utilization ; lipid ; nitrogen ; nutritional ecology ; water
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Description / Table of Contents: Résumé Peu de travaux ont examiné l'aptitude des insectes à modifier leur consommation et l'utilisation des aliments en fonction des variations qualitatives et quantitatives connues de l'aliment. On a rarement examiné si ces modifications sont adaptatives, si elles maintiennent le taux de croissance et le niveau des ressources, ou si elles limitent au moins les effets nuisibles du changement d'aliment. En utilisant une technique gravimétrique, nous avons mesuré le gain des poids, l'aliment consommé et les excréments produits par des chenilles de A. gemmatalis Hübn. (Lep, Noctuidea), important ravageur du soja aux USA. La consommation en poids frais (fw) a augmenté presque 2 fois quand le régime artificiel a été dilué progressivement avec de l'eau (65, 79, 86 ou 89% d'eau); le poids sec (dw) et le taux de consommation relative (RCR) ont diminué néanmoins avec la dilution. L'efficacité de digestion et d'assimilation de l'aliment consommé (digestibilité approchée, AD) a augmenté pour les trois dilutions, l'efficacité de conversion en biomasse de l'aliment digéré (ECD) a augmenté dans les dilutions à 79 et 85%, mais a diminué pour les régimes plus dilués. Par conséquent, le gain dw et le RGR, — produit de RCR x AD x ECD-, étaient identiques avec le régime à 79% fw à ceux de 65% fw, mais ont diminué pour les régimes plus dilués. Le taux relatif d'efficacité de consommation d'azote a diminué aussi avec la dilution, mais il a été compensé par un accroissement de l'efficacité de l'utilisation de l'azole tel, que le produit,-taux relatif d'accumulation de l'azote-, était le même pour les quatre régimes. La teneur en lipides des insectes a diminué, de 32% pour le régime sans dilution, à 13% pour le régime le plus dilué; elle a été le principal responsable de la diminution de RGR. L'accroissement de la consommation fw et de AD, tout en n'empêchant pas la diminution de RGR pour les deux régimes les plus dilués, a limité l'effet de la dilution (sans ces accroissements, le RGR du régime le plus dilué n'aurait été que 43% du RGR obtenu). Ces résultats indiquent que la consommation et l'utilisation d'aliments constituent un processus dynamique, et que les chenilles de A. gemmatalis comme beaucoup d'autres insectes, présentent des réactions compensatrices au changement de qualité de l'aliment. De telles variations dans la consommation d'aliments ont des conséquences écologiques (consommation accrue de substances allélochimiques potentiellement toxiques et exposition accrue aux causes biologiques de mortalité pendant l'alimentation), et intéressent l'organisation de la protection contre les insectes:-en permettant la manipulation de l'alimentation des insectes pour augmenter la consommation à court terme d'agents bioactifs, et, par ce moyen, réduire la consommation totale,-en permettant d'améliorer la modélisation des dégâts alimentaires prévisibles, qui, pour être efficace, devrait inclure les variations de l'alimentation en fonction de la qualité alimentaire de la plante cultiveé.
    Notes: Abstract Fresh weight (fw) food consumption by caterpillars of Anticarsia gemmatalis Hübner (Lepidoptera: Noctuidae) increased almost 2-fold as the nutrients in an artificial diet were increasingly diluted with water (diets contained 65, 79, 86 or 89% fw water). Nonetheless, dry weight (dw) relative consumption rate (RCR) declined with diet dilution. The efficiency at which the consumed food is digested and assimilated (approximate digestibility, AD) increased on the 3 diluted diets, and the efficiency at which digested food is converted to biomass (ECD) increased on the 79 and 86% fw diets. As a consequence, dw gained and relative growth rate (RGR), which is the product of RCR x AD x ECD, on the 79% fw diet were similar to those on the 65% fw diet, but they declined on the more diluted diets. Relative nitrogen consumption rate also declined with dietary dilution, but this was compensated by an increase in nitrogen utilization efficiency such that the product of these, relative nitrogen accumulation rate, was similar on all four diets. Insect lipid content declined from 32% on the undiluted diet to 13% dw on the most diluted diet, and was primarily responsible for the decline in RGR. The increases in fw consumption and AD, while not preventing a decline in RGR on the two most diluted diets, mitigated the impact of dietary dilution (e.g., without these increases, RGR on the most diluted diet would have been only 43% of that attained). These results indicate that the consumption and utilization of food are dynamic processes, and that caterpillars of A. gemmatalis, like many other insects, exhibit compensatory responses to changes in dietary quality.
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  • 5
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Site specific recombination ; 2 μ DNA plasmid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The purpose of this work is to identify and quantitate in vivo 2 μ plasmid FLP-independent recombination in yeast, using a nonselective assay system for rapid detection of phenotypic expression of the recombination events. A tester plasmid was constructed such that in vivo recombination between 2 μ direct repeat sequences produces the resolution of the plasmid into two circular DNA molecules. This recombinational event is detected as a phenotypic shift from red to white colonies, due to the mitotic loss of the plasmid portion containing the yeast ADE8 gene in a recipient ade1 ade2 ade8 genetic background. In the absence of the 2 μ FLP recombinase and/or its target DNA sequence, recombination is not abolished but rather continues at a high frequency of about 17%. This suggests that the FLP-independent events are mediated by the chromosomally-encoded general homologous recombination system. We therefore conclude that the totality of 2 μ DNA recombination events occurring in FLP+ cells is the contribution of both FLP-mediated and FLP-independent events.
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  • 6
    ISSN: 1432-0983
    Keywords: Schizosaccharomyces pombe ; Saccharomyces cerevisiae ; Argininosuccinate lyase ; Gene cloning
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A gene bank of Sau3A partially restricted Schizosaccharomyces pombe DNA in YEp13 was used to transform an arg4 mutant of Saccharomyces cerevisiae. One colony was recovered which contained the YEp13 plasmid bearing a large insert complementing the argininosuccinate lyase (ASL) mutation. As shown by restriction mapping and subcloning experiments, the DNA sequence required for complementation is localized on a 2 kb BamHI-BamHI fragment. The plasmid complemented several S. cerevisiae arg4 mutants of independent origin and a S. pombe arg7 mutant lacking ASL. Low but significant ASL activities were detected in crude extracts of these transformants. No complementation of the E. coli argH mutant was observed. Southern blot hybridizations showed that the insert originates from the S. pombe genome. No cross-hybridization was found between this sequence and S. cerevisiae DNA. It can be concluded that the cloned DNA fragment bears the S. pombe ARG7 gene coding for ASL.
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  • 7
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    Current genetics 14 (1988), S. 413-418 
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Chromosome length polymorphisms ; FIGE ; OFAGE
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Although Saccharomyces cerevisiae strains generally have similar chromosomal band patterns as revealed by pulsed field gel electrophoresis, individual bands often move slightly differently from one strain to the other. Surveying strains from our stock collection, we found that nearly all the bands of a certain pair of strains differed in their mobility. Some of these chromosome length polymorphisms segregated in a 2:2 ratio, indicating that they resulted from single structural alterations (i.e. additions or deletions). One of these was mapped on the right arm of chromosome 1. Others did not segrate in a simple 2:2 ratio. That is, there were progenies which had bands not present in either parent. We suggest that these new bands are the products of recombination between homologous chromosomes having two or more structural alterations.
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  • 8
    ISSN: 1432-0983
    Keywords: ARS ; Saccharomyces cerevisiae ; Schizosaccharomyces pombe ; Tetrahymena thermophila
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have isolated several Tetrahymena thermophila chromosomal DNA fragments which function as autonomously replicating sequences (ARS) in the heterologous Saccharomyces cerevisiae and Schizosaccharomyces pombe selection systems. The Tetrahymena ARS sequences were first isolated in S. cerevisiae and were derived from non-ribosomal micro- and macronuclear DNA. Sequence analysis of the ARS elements identified either perfect or close matches with the 11 by S. cerevisiae ARS core consensus sequence. Subcloning studies of two Tetrahymena ARS elements defined functional regions ranging in size from 50 to 300 bp. Testing of the ARS elements in S. pombe revealed that most of the T. thermophila inserts confer ARS function in both yeasts, at least in the sense of promoting a high transformation frequency to plasmids which contain them. However, the actual sequences responsible for ARS activity were not always identical in the two yeasts.
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  • 9
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Ribosomal protein genes ; CYH2
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A diploid strain of the yeast Saccharomyces cerevisiae has been constructed that has one copy of the ribosomal protein gene CYH2 completely deleted and replaced with the TRP1 gene using the method of Rothstein (1983). There are only small differences in growth rate and no detectable difference in steady state level of CYH2 mRNA between the diploid that is heterozygous for the CYH2 deletion and the parent diploid with two normal copies of this gene. This suggests that the diploid must partially compensate for the loss of one CYH2 gene. Tetrad dissection shows that haploid spores lacking the CYH2 gene cannot germinate. The lethality of this deletion can be rescued by a CYH2 cDNA on a low copy vector. Haploids which lack the genomic copy of the CYH2 gene, but contain a plasmid copy of the CYH2 cDNA are able to grow normally. These CYH2 deleted yeast haploids should be useful to analyze mutationally altered CYH2 genes and genes homologous to CYH2 from other organisms without interference from a genomic copy.
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  • 10
    ISSN: 1432-0983
    Keywords: Yeast ; Gene regulation ; Saccharomyces cerevisiae ; PDCI promoter
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A 870 by promoter fragment of the PDC1 gene that includes the carbon source dependent regulatory regions was investigated using 5′ and 3′ promoter deletions. The results indicate that glucose and ethanol regulation of PDC1 transcription are independently controlled by distinct cis-acting regions. The consensus sequence AAATCGATA may play a role in this regulation, while the sequence (ATCA)AACCT may be important in transcription initiation.
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  • 11
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Vector ; Glyphosate resistance ; Transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The E. coli aroA gene was inserted between yeast promoter and terminator sequences in different shuttle expression plasmids and found to confer enhanced EPSP synthase activity as well as resistance to glyphosate toxicity. Subsequently, a transformation system using these newly constructed vectors in yeast was characterized. The efficiency of the glyphosate resistance marker for transformation and selection with plasmid pHR6/20-1 in S. cerevisiae laboratory strain SHY2 was found to be relatively high when compared with selection for LEU2 prototrophy. The fate of the recombinant plasmid pHR6/20-1 in the transformants, the preservation of the aroA E. coli DNA fragment in yeast, mitotic stability, EPSP synthase activity, and growth on glyphosate-containing medium have been investigated. As this plasmid also allows direct selection for glyphosate resistant transformants on rich media, the glyphosate resistance marker was used for transforming both S. cerevisiae laboratory strain SHY2 and brewer's yeast strains S. cerevisiae var. “uvarum” BHS5 and BHS2. In all cases, the vector pHR6/20-1 was maintained as an autonomously replicating plasmid. The resistance marker is, therefore, suitable for transforming genetically unlabeled S. cerevisiae laboratory, wild, and industrial yeast strains.
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  • 12
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Chromosome V ; Ty elements ; tRNA genes ; Transposition hot-spots ; Yeast polymorphism
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Ty4 is a novel transposable element in the yeast, Saccharomyces cerevisiae, which is present in only a few copies in the genome (Stucka et al. 1989). In strain C836 one of the three copies (Ty4-90) is contained in cosmid clone c90, where it resides on chromosome V. Analysis of this region reveals a “hot-spot” of transposition: in addition to Ty4-90, the locus contains a complete Ty3 element and seven singular delta, sigma and tau elements. Three tRNA genes (for His, Lys, and Ile) are located in this region, and these are closely associated with one or the other of the elements, a phenomenon commonly observed in yeast. A comparison of c90 with corresponding regions from other strains shows that the locus is highly polymorphic and that this polymorphism is explicitly associated with Ty transposition and recombination events.
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  • 13
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    Current genetics 16 (1989), S. 315-321 
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Antisuppressor ; ADE3 ; Translation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Mutations in a known yeast gene, ADE3, were shown to act as an antisuppressor, reducing the efficiency of the omnipotent suppressor, sup45-2. The ADE3 locus encodes the trifunctional enzyme C1-tetrahydrofolate synthase, which is required for the biosynthesis of purines, thymidylate, methionine, histidine, pantothenic acid and formylmethionyl-tRNAfmet. The role of this enzyme in translational fidelity had not previously been suspected.
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  • 14
    ISSN: 1432-0983
    Keywords: Yeast ; Saccharomyces cerevisiae ; Nonsense suppression ; Omnipotent suppressors ; Gene mapping
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Ten dominant omnipotent suppressors of Saccharomyces cerevisiae, which were previously shown to be different from SUP46, have been examined. Nine are mapped in a region between lys5 and cyh2 on the left arm of chromosome VII. These suppressors, like SUP46, manifest sensitivity to increased temperature and the antibiotics paromomycin and hygromycin B. In addition, they have an identical action spectrum. These results strongly suggest that they are allelic to each other and they are designated SUP138. The tenth is mapped to a position between his1 and arg6 on the right arm of chromosome V. This suppressor, named SUP139, does not manifest temperature sensitivity nor antibiotic sensitivity. SUP139 and SUP138, which are clearly distinguished by means of action spectrum, act on much fewer nonsense mutations than SUP46. It is now clear that dominant omnipotent suppressors arising at a single locus are homogeneous and that their efficiency is locus-dependent. The order of efficiency is SUP46〉SUP138〉SUP139.
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  • 15
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    Current genetics 13 (1988), S. 21-23 
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Transformation ; Plasmid ; Colony ; Polyethylene glycol
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A rapid and simple yeast transformation procedure has been developed using colonies on agar plates. Saccharomyces cerevisiae SHY3 cells were picked up from colonies on YPD plates grown freshly or stored at 4 °C and incubated with M13RK9-T DNA at 30 °C for 1–2 h in a solution of Li+, Ca2+, Mg2+, triacetin and polyethylene glycol. About 3,500 transformants were obtained per µg of double stranded M13RK9-T DNA. Unlike the existing spheroplast techniques, single stranded M13RK9-T DNA transformed intact cells below one-hundredth frequency of the duplex form.
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  • 16
    ISSN: 1432-0983
    Keywords: Aspergillus terreus clonotheque ; Saccharomyces cerevisiae ; Homologous integration ; 2 μ circledirected chromosome destabilization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A genome clonotheque consisting of 25- to 40-kb Sau3A1 fragments of Aspergillus terreus DNA was constructed in the episomal cosmid vector pES33 containing the yeastARG4 gene. From the 475 transformants of cir° yeast strain ESH-0, 23 stable Arg + transformants were independently selected. Genetic and Southern analysis of these stable transformants showed that 39% arose as a result of recombination between cloned A. terreus DNA sequences and yeast chromosome XII. The recombination events most likely occurred in the regions of homology within the rDNA clusters of A. terreus and Saccharomyces cerevisiae.
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  • 17
    ISSN: 1432-0983
    Keywords: Yeast ; Repair ; Complementation ; Saccharomyces cerevisiae ; Schizosaccharomyces pombe ; Gene cloning
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Two Saccharomyces cerevisiae genes necessary for excision repair of UV damage in DNA, RAD1 and RAD2, were introduced individually, on a yeast shuttle vector, into seven Schizosaccharomyces pombe mutants — rads1, 2, 5, 13, 15,16 and 17. The presence of the cloned RAD1 gene did not affect survival of any of the S. pombe mutants. The RAD2 gene increased survival of S. pombe rad13 to near the wild-type level after UV irradiation and had no effect on any of the other mutants tested. S. pombe rad13 mutants are somewhat defective in removal of pyrimidine dimers so complementation by the S. cerevisiae RAD2 gene suggests that the genes may code for equivalent proteins in the two yeasts.
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  • 18
    ISSN: 1432-0983
    Keywords: DNA transformation ; Saccharomyces cerevisiae ; Site-specific recombination ; 2μ DNA plasmid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The 2μ DNA plasmid of the yeast Saccharomyces cerevisiae does not confer any known selectable phenotype to the host cell carrying it. Selection of cells transformed with purified 2μ DNA therefore cannot be achieved, and the intracellular presence of 2μ can only be assessed by molecular analysis of the DNA complement. In addition, 2μ alone does not replicate in bacterial hosts, thus rendering its amplification by conventional methods impossible. We have isolated a shuttle plasmid, pBH-2L, generated by in vivo sites-pecific recombination between the endogenous 2μ DNA plasmid and pRL, a pBR322 derivative containing the yeast LEU2 gene and one 2μ repeat sequence associated with the origin of replication. This new shuttle plasmid has the property, when transformed into yeast, of undergoing site-specific recombinational resolution between its two direct repeat sequences. This releases 2μ plasmid and pRL as individual molecules. The latter can undergo progressive mitotic loss during growth in nonselective medium, ultimately leaving leucine auxotrophic transformants that contain only 2μ DNA plasmid. This system can be utilized to introduce 2μ DNA alone into cells lacking it, thereby providing a novel means to study the biology and the molecular genetics of the plasmid and its potential practical applications as a vector.
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  • 19
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    Current genetics 15 (1989), S. 113-120 
    ISSN: 1432-0983
    Keywords: Calmodulin mutant ; Nuclear division ; Chromosome stability ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The coding region of a yeast calmodulin gene was fused to a galactose-inducible GAL1 promoter, and a conditional-lethal mutant of Saccharomyces cerevisiae, in which the expression of calmodulin was regulated by galactose, was constructed. The mutant grew normally in galactose medium, but in glucose medium, in which the promoter was repressed, it ceased growing after 12–15 h. The growth arrest was associated with a decrease in intracellular calmodulin levels: after 12h, no intracellular calmodulin protein was detectable. Analysis of the terminal phenotype showed that when the cell stopped growing, it had a bud, a nucleus after S-phase and a short mitotic spindle. Thus, the defect was mainly in nuclear division. Bud growth was partially inhibited in these cells: 27% of the cells stopped growing with a small bud. Furthermore, calmodulin-deficient cells showed elevated rates of chromosome loss, possibly as the result of a defect in the precise segregation of chromosomes.
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  • 20
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    Current genetics 15 (1989), S. 235-237 
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Mitochondrial DNA ; Method of extraction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A rapid method for the extraction of yeast mitochondrial DNA (mtDNA) is described. In comparison with previous methods, it simplifies several steps, does not require either the isolation of mitochondria or phenol treatment and is less time consuming. This protocol gives a high yield of pure mtDNA (50–120 μg from a 100-ml culture), which can be directly used in various molecular applications: restriction enzyme digestion, electrophoresis, blotting, labeling, cloning and sequencing.
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  • 21
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Mutagen hyperresistance ; Southern, Northern analysis ; Gene transplacement ; Transposon mapping
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The genes SNQ and SFA confer hyperresistance to 4-NQO and FA when present on a multi-copy plasmid in yeast. Both are non-essential genes since transplacement of SNQ by a disrupted snq-0::LEU2 yielded stable and viable haploid integrants. Southern analysis revealed that SNQ and SFA are single-loci genes, and OFAGE analysis showed that they are located on chromosome XIII and IV, respectively. Northern blot analysis of SNQ and SFA revealed poly(A)+ RNA transcripts of 2 kb and 1.7 kb, respectively. Nuclease S 1 mapping showed SNQ to have a coding region of 1.6 kb and SFA, one of 1.3 kb. The 5′ coding regions were determined for both genes, while the 3′ end could only be determined for gene SNQ. Both genes do not appear to contain introns. The SFA locus was also mapped by transposon mutagenesis. Tn10-LUK integrants disrupted the SFA gene function at sites that were determined by subcloning to lie within the SFA transcription unit.
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  • 22
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; SKI3 ; SKI5 ; M1 dsRNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have identified a mutant strain of the yeast Saccharomyces cerevisiae which overproduces killer toxin. This strain contains a single mutation which fails to complement defects in both the SKI3 and SKI5 genes, while a cloned copy of this gene complements both the ski3 and ski5 defects. The level of secreted toxin from a cDNA based plasmid is not increased in a ski3 strain, showing that the overproduction phenotype is dependent upon an increased level of M1 dsRNA.
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  • 23
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Psoralen photoaddition ; Interstrand cross-link ; Repair deficiency ; Genotoxicity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Two different UVA irradiation systems were initially biologically calibrated with two haploid yeast strains proficient and deficient, respectively, in nucleotide excision repair. The number of DNA lesions introduced into the cell's genome by the photoactivated bifunctional furocoumarin 8-MOP was then calculated by means of the applied UVA exposure doses. At LD37 the repair-proficient wild type had about 14 ICL and 34 furan-side monoadducts in its DNA, while doubly blocked repair mutant rad3-12 pso1-1 had 2 ICL and 3 monoadducts. Locus-specific reversion of lys1-1 followed two-hit kinetics in the repair-proficient wild type and one-hit kinetics in an excision-deficient rad2-20 mutant, as would be expected if ICL was the main type of mutagenic lesion in the wild type and monoadducts the main mutagenic lesion type in the excision-deficient strain. Quantitative comparison of 8-MOP + UVA-induced ICL with those induced by bifunctional mustard revealed the former to have a much higher genotoxicity.
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  • 24
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    Current genetics 13 (1988), S. 283-289 
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Inducible antisense gene ; Acetolactate synthase ; Bradytrophic phenocopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We report for the first time on the regulation of gene expression in yeast by antisense RNA. Chimaeric genes were constructed containing the 5′ upstream and partial coding sequence of SMR1 — a sulfometuron methyl resistant allele of the ILV2 locus. Such fragments were placed 5′ to 3′ and 3′ to 5′ under control of the GAL10 promoter and CYCl terminator in a high copy YEp plasmid. Following galactose induction only transformants containing antisense RNA genes showed biological activity against SMR1 gene expression. Antisense RNA inhibited synthesis of the SMR1 gene product acetolactate synthase and thus repressed cellular growth which resulted in a bradytrophic auxotroph revertable by addition of isoleucine and valine. Antisense RNA inhibition was enhanced in galactose medium containing sulfometuron methyl and in gcn4 cells deficient for positive regulation of the ILV2 locus. This system can be used to study factors that interfere with antisense RNA function and to assign biological function to randomly cloned DNA fragments.
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  • 25
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    Current genetics 14 (1988), S. 331-335 
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Meiosis ; Deletion mutations ; Sequence dissimilarities
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A diploid yeast strain with extensive sequence dissimilarity in homologous regions near the LYS2 locus was sporulated, and spontaneous lys2 and lys5 mutant spores, selected on α-amino adipate, were analyzed. As many as 50% of the mutant spores contained a deletion in LYS2. These deletions occurred at a frequency of 5.0 × 10−7. While deletions of various sizes and endpoints were obtained, all the deletions recovered in this study included the border between homologous and non-homologous sequences located 4 kb upstream of LYS2. Large lys2 deletions that extended into an adjacent CYH2 duplication occurred at a frequency of 2.0 × 10−7, more than 1,000 times the frequency of the CYH2-LYS2 deletions found in a related haploid strain. This high frequency of CYH2-LYS2 deletions was observed only after sporulation of the diploid strain, and was dependent upon extensive sequence dissimilarity near the LYS2 locus.
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  • 26
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Allosuppressor ; Translation ; Fidelity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Allosuppressor (sal) mutations enhance the efficiency of the yeast ochre suppressor SUQ5 and define five unlinked loci, SALT-SALS. A number of sal4 mutants were isolated and found to have pleiotropic, allele;specific phenotypes, including hypersensitivity in vivo to paromomycin and other antibiotics that stimulate translational errors in yeast. To examine further the nature of the SAL4 gene product, the wild type SAL4 gene was isolated by complementation of a conditional lethal allele sal4-2, and demonstrated to be a single copy gene encoding a single 1.6 kb transcript. Restriction mapping and DNA hybridisation analysis were used to demonstrate that the SAL4 gene is identical to the previously identified omnipotent suppressor gene SUP45 (SUPT). Our results implicate the SAL4 gene product as playing a major role in maintaining translational accuracy in yeast.
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  • 27
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Protoplast fusion
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    Topics: Biology
    Notes: Summary The percentage of hybrids formed during protoplast fusion in Saccharomyces cerevisiae is determined by the percentage of protoplasts at the GI/S boundary of the cell cycle.
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  • 28
    ISSN: 1432-0983
    Keywords: Peroxisomes ; Protein import ; Saccharomyces cerevisiae ; Hansenula polymorpha
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The DAS gene of Hansenula polymorpha was expressed in Saccharomyces cerevisiae under the control of different promoters. The heterologously synthesized dihydroxyacetone synthase (DHAS), a peroxisomal enzyme in H. polymorpha, shows enzymatic activity in baker's yeast. The enzyme was imported into the peroxisomes of S. cerevisiae not only under the appropriate physiological conditions for peroxisome proliferation (oleic acid media), but also in glucose-grown cells where it induced the enlargement of the few peroxisomes present. This growth process was not accompanied by an increase in the number of microbodies, which suggests a separate control mechanism for peroxisomal proliferation.
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  • 29
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    Current genetics 15 (1989), S. 399-401 
    ISSN: 1432-0983
    Keywords: Saccharomyces exiguus ; Saccharomyces cerevisiae ; HO gene ; MAT gene
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The DNA of Saccharomyces exiguus was analyzed by Southern hybridization using cloned MATa, MATα, and HO genes of Saccharomyces cerevisiae as probes. It was shown that S. exiguus has a DNA sequence homologous with the HO gene of S. cerevisiae and that this DNA sequence is on a chromosome of about 940 kb of DNA in S. exiguus. However, there is no DNA sequence in S. exiguus that is homologous with the MAT genes of S. cerevisiae.
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  • 30
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    Nutrient cycling in agroecosystems 15 (1988), S. 173-179 
    ISSN: 1573-0867
    Keywords: Douglas-fir ; urea ; basal area increment ; growth response ; nitrogen ; critical level
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract A series of fourteen Pacific North-west Douglas-fir installations, ranging in age from 6 to 26 years were analysed with respect to site factors, foliage nutrients, and growth response to applied fertilizer. Unfertilized basal area increment ranged from 1.2 to 3.1 m2 ha−1 yr−1 with no apparent relationship with soil, stand age or site index. Basal area increment was correlated with foliage N and a critical level for N was calculated as 1.7%. Applications of 220 kg N ha−1 as urea increased growth between 0 and 95% of the unfertilized basal area growth, with an average of 24.9%. Response could be predicted from foliage N and unfertilized basal area increment. When the same relationships were applied to previously older stand data, results were more variable as elements such as B and S showed evidence of being limiting.
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  • 31
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    Nutrient cycling in agroecosystems 15 (1988), S. 203-210 
    ISSN: 1573-0867
    Keywords: irrigation ; pineapples ; nitrogen ; potassium ; yield ; Nigeria
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The investigation studied the effect of fertilization with four nitrogen levels (0, 100, 150 and 200 kg/ha), four potassium levels (0, 100, 150 and 200 kg/ha) and four irrigation treatments (0, 3, 7 and 14-day interval) on the growth and yield of smooth Cayenne pineapples. Increasing irrigation frequency increased growth parameters of number of leaves, D-leaf length and days to 50% flowering. Fruit weight was highest at N = 150 kg/ha, K = 200 kg/ ha and irrigation once a week. Higher N levels (beyond 150 kg/ha) applied after the plant crop has been harvested did not further enhance yields of the ratoon crop. For K = 200 kg/ha potential and harvested yields were reduced in all irrigation treatments when N 〉 150 kg/ha.
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  • 32
    ISSN: 1573-0867
    Keywords: farmyard manure ; maize ; nitrogen ; phosphorus ; rice ; wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Field experiments with rice-wheat rotation were conducted during five consecutive years on a coarse-textured low organic matter soil. By amending the soil with 12t FYM ha−1, the yield of wetland rice in the absence of fertilizers was increased by 32 per cent. Application of 80 kg N ha−1 as urea could increase the grain yield of rice equivalent to 120 kg N ha−1 on the unamended soil. Although the soil under test was low in Olsen's P, rice did not respond to the application of phosphorus on both amended and unamended soils. For producing equivalent grain yield, fertilizer requirement of maize grown on soils amended with 6 and 12 t FYM ha−1 could be reduced, respectively to 50 and 25 per cent of the dose recommended for unamended soil (120 kg N + 26.2 kg P + 25 kg K ha−1). Grain yield of wheat grown after rice on soils amended with FYM was significantly higher than that obtained on unamended soil. In contrast, grain yield of wheat which followed maize did not differ significantly on amended or unamended soils.
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  • 33
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    Nutrient cycling in agroecosystems 18 (1988), S. 245-250 
    ISSN: 1573-0867
    Keywords: plantain ; intercropping systems ; nitrogen
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The investigation evaluated the productivity of plantain intercropped with cassava, cocoyam and yam, fertilized annually with 0, 320 and 480 kg N ha−1 respectively. Yields from nitrogen fertilized intercrops were higher than those of unfertilized treatments. In plantain + cassava intercrop receiving 480 kg N ha−1 plantain growth was suppressed. Plantain intercropped with yam and fertilized with 320 kg N ha−1 matured early and produced better bunches than other treatments. Plantain + yam or cocoyam intercropping systems fertilized with 320 kg N ha−1 were recommended because of improved plantain establishment and increased combined crop yields.
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  • 34
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    Pure and applied geophysics 127 (1988), S. 529-544 
    ISSN: 1420-9136
    Keywords: Ionosphere ; nitrogen ; vibrational temperatures ; N2 and CO2 and populations of N2 and CO2 in the D, E and F regions
    Source: Springer Online Journal Archives 1860-2000
    Topics: Geosciences , Physics
    Notes: Abstract Theoretical and experimental aspects of the production, transformation, diffusion and loss of N2 in the upper atmosphere are considered. The N2-CO2 near-resonant system in theD andE regions is taken into account. We describe our understanding of the methods necessary to find the vibrational populations of N2 and CO2 (asymmetric mode of CO2). The calculations of the vibrational temperatures in theD, E, andF regions for the mid-latitude ionosphere and an aurora are presented. The connection between the excited species and the 4.26-μm radiation intensities is considered. The models for the rate coefficient of the reaction of O+ with N2 and the electron density decrease resulting from N2 in the F region are discussed.
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  • 35
    ISSN: 1420-9071
    Keywords: Saccharomyces cerevisiae ; growth conditions ; kinaseless mutant ; plasma membrane vesicles ; glucose transport ; kinetics and computer simulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In this study experimental data on the kinetic parameters investigated by other authors1–5, 11 together with own data on plasma membrane vesicles, have been subjected to a computer simulation based on the equations describing facilitated diffusion. The simulation led to an ideal fit describing the above data. From this it can be concluded that glucose is transported by facilitated diffusion, and not by active transport as was postulated by Van Steveninck14, 15. The simulation method also demonstrates that the fast sampling technique used by these authors1–5,11 underestimates the fluxes. Thus, the parameters given do not contribute to the understanding of glucose transport under different metabolic conditions. The K value of plasma membrane vesicles prepared from glucose-repressed cells is around 7 mM. Derepression, particularly by galactose, causes a highly significant increase in affinity as shown by a decrease in the K value to 2 mM. The highest affinity was measured in a triple kinaseless mutant grown on glycerol with a K value of 1 mM. If seems, therefore, that the kinetic parameters derived from initial uptake rates of glucose in intact cells1–5,11 using single flux analysis, such as Eadie-Hofstee- or Lineweaver-Burk-plots, are in error.
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  • 36
    ISSN: 1432-0983
    Keywords: Formaldehyde ; DNA-protein cross-links ; Repair ; Saccharomyces cerevisiae ; Hyperresistance
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The formation and removal of formaldehyde-mediated DNA protein cross-linking was measured by CsCI density gradient analysis in yeast strains of differing resistance to formaldehyde. Wild-type cells and transformants made hyperresistant to formaldehyde by a multi-copy vector containing the yeast SFA gene were specifically labeled in their DNA and incubated in the presence of formaldehyde. Treatment with formaldehyde lead to the formation of equal amounts of DNA protein cross-links; subsequent liquid holding of cells for 24 h resulted in the removal of nearly all DNA protein crosslinks regardless of the original formaldehyde resistance status of the strains.
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  • 37
    ISSN: 1432-072X
    Keywords: Saccharomyces cerevisiae ; Yeast ; Phospholipase B ; Lysophospholipase ; Enzyme inhibition ; AMP ; Unesterified fatty acids
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    Topics: Biology
    Notes: Abstract Divalent cations activate the lysophospholipase and transacylase reactions catalyzed by the same enzymes in the yeast Saccharomyces cerevisiae. The activation was observed at neutral pH, but not at the pH optimum of lysophospholipase/transacylase, near 3.5. Adenine nucleotides, especially AMP and ADP, are strong inhibitors of the same group of enzymes. Half maximal inhibition by AMP was found at a concentration of about 20 μM. The inhibition by nucleotides in low concentrations is enhanced by divalent cations.
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  • 38
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    Archives of microbiology 151 (1988), S. 20-25 
    ISSN: 1432-072X
    Keywords: Yeast ; Saccharomyces cerevisiae ; Mating ; Zygote formation ; Chloroquine ; Lysosomotropic agent ; Plasma membrane ; Cell fusion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Haploid cells of opposite mating type of Saccharomyces cerevisiae conjugate to form zygote. During the conjugation process, the degradation or reorganization of the cell wall and the fusion of the two plasma membranes take place. Since chloroquine inhibits cellular events associated with the reorganization of the plasma membrane, the effect of the drug on conjugation was studied. Chloroquine at a concentration, at which cell growth was not retarded, inhibited zygote formation, while it did not affect other mating functions, such as sexual agglutination, production of and response to mating pheromone. Cells in a mating culture containing chloroquine formed no “prezygote” suggesting that they were not prepared for entering into fusion process. The inhibitory effect of chloroquine was reversible as cells formed zygote when they were washed after treatment with chloroquine. Zygote formation was unaffected in cells possessing chlorquine within vacuoles after incubation with the drug in complete medium (YPD) at pH 7.5, followed by washing. This suggests that chloroquine inhibits zygote formaton by adsorbing to the plasma membrane of S. cerevisiae.
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  • 39
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    Archives of microbiology 150 (1988), S. 313-319 
    ISSN: 1432-072X
    Keywords: 3′-Phosphoadenylyl sulphate reductase ; Sulphite formation ; Cysteine biosynthesis ; Thioredoxin ; Saccharomyces cerevisiae ; HPLC enzyme analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The enzymatic mechanism of sulphite formation in Saccharomyces cerevisiae was investigated using a purified 3′-phosphoadenylsulphate (PAPS) reductase and thioredoxin. The functionally active protein (MR 80–85 k) is represented by a dimer which reduces 3′-phosphoadenylyl sulphate to adenosine-3′,5′-bisphosphate and free sulphite at a stoichiometry of 1:1. Reduced thioredoxin is required as cosubstrate. Examination of the reaction products showed that free anionic sulphite is formed with no evidence for “bound-sulphite(s)” as intermediate. V max of the enriched enzyme was 4–7 nmol sulphite · min-1 · mg-1 using the homologous thioredoxin from yeast. The velocity of reaction decreased to 0.4 nmol sulphite · min-1 · mg-1 when heterologous thioredoxin (from Escherichia coli) was used instead. The K m of homologous thioredoxin was 0.6 · 10-6 M, for the heterologous cosubstrate it increased to 1.4 · 10-6 M. The affinity for PAPS remained practically unaffected (K m PAPS: 19 · 10-6 M in the homologous, and 21 · 10-6 M in the heterologous system). From the kinetic data it is concluded that the enzyme followed an ordered mechanism with thioredoxin as first substrate followed by PAPS as the second. Parallel lines in the reciprocal and a common intersect in the Hanes-plots for thioredoxin were seen as indication of a ping-pong (with respect to thioredoxin) uni-bi (with respect to PAPS) mechanism.
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  • 40
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    Archives of microbiology 151 (1989), S. 198-202 
    ISSN: 1432-072X
    Keywords: Sexual agglutination ; Mating ; Saccharomyces cerevisiae ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Genetic regulation of the inducibility of sexual agglutination ability in the yeast Saccharomyces cerevisiae was studied. Detailed analysis of the degree of sexual agglutination was carried out; it showed that a greater number of genes are involved in the regulation of inducible sexual agglutination in strain H1-0 than previously assumed. Although dominancy of inducible phenotype over constitutive was confirmed, the effectiveness of one gene changing the constitutive phenotype to the inducible seemed to be somewhat low. Quantity per cell of agglutination substances responsible for sexual agglutination increased as the agglutination ability became greater.
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  • 41
    ISSN: 1432-072X
    Keywords: Yeast ; Saccharomyces cerevisiae ; Mating reaction ; Zygote formation ; Mating pheromone ; Fatty acid ; Arachidonic acid
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    Topics: Biology
    Notes: Abstract Effect of exogenous fatty acids on zygote formation in Saccharomyces cerevisiae was studied. Arachidonic and oleic acids considerably stimulated zygote formation, but other fatty acids tested, linoleic, linolenic, stearic and palmitic acids, did not. Pretreatment experiments with arachidonic acid showed that the stimulation of zygote formation by the fatty acid required the presence of mating pheromone.
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  • 42
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    Archives of microbiology 151 (1989), S. 391-398 
    ISSN: 1432-072X
    Keywords: Saccharomyces cerevisiae ; Exoglucanases ; Purification ; Protein moieties ; Tunicamycin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Exoglucanase (exo-1,3-β-D-glucan glycohydrolase, EC 3.2.1.56) activity secreted by Saccharomyces cerevisiae into the culture medium was separated by ion exchange chromatography into two glycoprotein isoenzymes which contributed 10% (exoglucanase I) and 90% (exoglucanase II) towards the total activity. Analysis of the “in vitro” deglycosylated products by polyacrylamide gel electrophoresis under native or denaturing conditions indicated that the protein portions of both exoglucanases exhibited identical mobility, each one consisting of two polypeptides with M r of 47000 and 48000. The same profile was shown by the exoglucanase secreted in the presence of tunicamycin. Antibodies raised against the protein portion of exoglucanase II did react with both native exoglucanases and their deglycosylated products with a pattern indicative of immunological identity. Digestion of the “in vitro” deglycosylated products of both exoglucanases with Staphylococcus aureus V-8 protease or trypsin generated the same proteolytic fragments in each case. Only exoglucanase II was secreted by protoplasts. These and previously reported results indicate that the protein portions of both isoenzymes may be the product of the same gene (or a family of related genes), and that exoglucanase I is a product of enzyme II, modified by a process occurring beyond the permeability barrier of the cell.
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  • 43
    ISSN: 1432-072X
    Keywords: Saccharomyces cerevisiae ; Recombination ; Tryptophan cluster ; Yeast vectors ; Plasmid copy number
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We describe a convenient method for the in vivo construction of large plasmids that possess a multitude of restriction sites. A large (23 kbases) circular self-replicating plasmid carrying a partial LEU2-d gene was cotransformed with a circular non-replicating plasmid carrying the entire LEU2 gene. In vivo recombination results preferentially in a plasmid that carries both the LEU2-d and the entire LEU2 gene. In addition we also found one plasmid with a tandem LEU2 insertion and one plasmid where the LEU2-d gene was replaced by the entire LEU2 gene.
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  • 44
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    Journal of industrial microbiology and biotechnology 2 (1988), S. 365-372 
    ISSN: 1476-5535
    Keywords: Osmotic pressure ; Intracellular ethanol ; Yeast ; Nutrient ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary The intracellular accumulation of ethanol in yeast and its potential effects on growth and fermentation have been topics of controversy for the past several years. The determination of intracellular ethanol based on the exclusion of [14C]sorbitol to estimate aqueous cell volume was used to examine the question of intracellular ethanol accumulation. An intracellular accumulation of ethanol inSaccharomyces cerevisiae was observed during the early stages of fermentation. However, as fermentation continued, the intracellular and extracellular concentrations of ethanol became similar. Increasing the osmotic pressure of the medium with glucose or sorbitol was observed to cause an increase in the intracellular ethanol concentration. Associated with this was a decrease in yeast growth and fermentation rates. In addition, increasing the osmotic pressure of the medium was observed to cause an increase in glycerol production. Supplementation of the media with excess peptone, yeast extract, magnesium sulfate and potassium phosphate was found to relieve the detrimental effects of high osmotic pressure. Under these conditions, though, no effect on the intracellular and extracellular ethanol distribution was observed. These results indicate that nutrient limitation, and not necessarily intracellular ethanol accumulation, plays a key role during yeast fermentations in media of high osmolarity.
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  • 45
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    Journal of chemical ecology 15 (1989), S. 2379-2390 
    ISSN: 1573-1561
    Keywords: Acidic fog ; Phaseolus lunatus ; Trichoplusia ni ; cabbage looper ; Lepidoptera ; Noctuidae ; nitrogen ; free amino acid ; soluble protein ; plantinsect interactions ; air pollution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Phaseolus lunatus L. (Henderson Bush lima beans) were exposed to 2 hr acidic fogs with 2.5∶1.0 (v/v) nitrogen-sulfur ratio typical of the west coast of the United States. Fogs with pH values of 2.0 (P 〈 0.01,t tests), 2.5 (P 〈 0.05), or 3.0 (P 〈 0.01) increased percent total nitrogen (dry weight) of foliage as compared to plants subjected to control fogs with a pH of 6.3–6.5. Fresh weight concentrations of soluble protein and certain free amino acid concentrations were increased by plant exposure to acidic fogs with a pH of 2.5 (t tests,P 〈 0.05). Concentrations of free amino acids considered essential for insect growth, as well as nonessential and total free amino acids were not significantly affected by any treatment (P 〉 0.05,t test). Water content (%) of foliage was not changed significantly (P 〉 0.05,t test) by exposure to any of the fogs.Trichoplusia ni (Hübner) (Lepidoptera: Noctuidae) larvae ate significantly more foliage and gained significantly more weight on plants treated with 3.0 pH fogs (P 〈 0.01,t test). Several potential explanations are offered for the lack of significant weight gain by larvae on plants in which soluble protein levels, free amino acid concentrations, or percent total nitrogen contents were enhanced by acidic fogs with a pH of 2.5 and 2.0. No larval feeding preference was detected for foliage exposed to acidic versus control fogs, and no significant differences were detected in percent survival ofT. ni eggs exposed to acidic or control fogs. Some implications of acidic fogs for population dynamics ofT. ni are discussed.
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  • 46
    ISSN: 1432-1424
    Keywords: vacuole ; lipid bilayer ; K-channel ; single channel ; DIDS ; yeast ; Saccharomyces cerevisiae ; Ca2+ activation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Summary A voltage-dependent and Ca2+-activated cation channel found in the vacuolar membrane of the yeast,Saccharomyces cerevisiae, was incorporated into planar lipid bilayer and its gating characteristics were studied at the macroscopic and single-channel levels. The open-channel probability at steady state, which was estimated by the macroscopic current measurement, gave a maximum value at −10 mV and decreased in a graded fashion as the voltage became more positive or more negative. The steady-state voltage dependence was explained by assuming two independent gates, which had different rate constants and opposite voltage dependence. The fast-responding gate opened when the voltage of thecis side (the side to which the vesicles were added) was made more negative and the slow-responding gate behaved in the opposite direction. Relatively high concentrations of Ca2+, about 1mm, were required on thecis side for opening the slow gate in a voltage-dependent manner. DIDS increased the open-channel probability of the fast gate when added to thecis side, but was ineffective on the slow gate.
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  • 47
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    Molecular genetics and genomics 211 (1988), S. 88-94 
    ISSN: 1617-4623
    Keywords: CupI ; Gene amplification ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Carcinogen-induced amplification at the CupI locus, coding for a metallothionein protein, was studied in the yeast Saccharomyces cerevisiae. Exposure of cells from three different haploid strains, 4939, DBY746 and 320, to chemical carcinogens such as N-methyl-N′-nitro-N-nitrosoguanidine (MNNG), ethylmethanesulfonate (EMS) and 4-nitroquinoline-N-oxide (4NQO) enhanced the frequency of copper-resistant colonies up to several hundred fold. Copper-resistant clones obtained from strains DBY746 and 320, which contain more than one copy of the CupI locus, displayed a four-to eightfold amplification of the CupI sequences. In these clones the amplified CupI sequences were organized in a tandem array. Carcinogen treatment of strain 4939 in which only one copy of the CupI gene is present produced resistant colonies without CupI amplification. The possible use of the yeast system to study gene duplication and amplification is discussed.
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  • 48
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    Molecular genetics and genomics 211 (1988), S. 446-454 
    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; Invertase genes ; Promoter sequences ; Palindromes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In the yeast Saccharomyces cerevisiae six unlinked structural genes for invertase, the SUC genes, are known. We sequenced about 800 bp of the 5′ non-coding region and the first 220 bp of the coding region of the genes SUC1, SUC3, SUC4 and SUC5 and compared them with the previously sequenced genes SUC2 and SUC7 (Sarokin and Carlson 1985a). All are highly homologous within the coding region but in the non-coding region SUC1 shows some differences and SUC2 is more highly diverged. Two different kinds of TATA boxes were identified: the more strongly expressed genes SUC1, 2 and 4 have the sequence TATAAA and the more weakly expressed genes SUC3, 5 and 7 have TACAAA. Though the SUC1 sequence is in general more homologous to the other SUC genes, the region between-140 and + 100 of SUC1 is nearly identical to SUC2. This could be due to a gene conversion between SUC1 and the silent suc2 o allele which occurs in the strains carrying SUC1. Within the upstream regions of all the SUC genes three regions with palindromic sequences analogous to stem and loop structures were identified. Comparable structure could be detected in similar positions in the upstream sequences of the divergently transcribed yeast gene pairs MAL6S-MAL6T and GAL1-GAL10. Implications for the importance of these structures in the regulation and initiation of transcription are discussed.
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  • 49
    ISSN: 1617-4623
    Keywords: l-azetidine-2-carboxylate resistance ; Escherichia coli ; γ-glutamyl kinase ; Proline ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We constructed plasmids carrying the Escherichia coli proB gene that encodes γ-glutamyl kinase, under the control of the yeast GAL1 promoter. This construction was carried out with both the wild-type proB + gene and a mutant allele, proB74, that specifies an enzyme resistant to feedback inhibition by proline. Yeast pro1 mutants harboring these plasmids are proline prototrophs. We conclude that the pro1 mutation results in a deficiency in the γ-glutamyl kinase activity in Saccharomyces cerevisiae. Expression of the proB74 allele in yeast resulted in enhanced resistance to the proline analogue l-azetidine-2-carboxylate and in a 2.4-fold elevation of the intracellular free proline levels. This result suggests that γ-glutamyl kinase is the rate limiting step in proline biosynthesis in yeast.
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    Molecular genetics and genomics 214 (1988), S. 249-256 
    ISSN: 1617-4623
    Keywords: Bent DNA ; DNA structure ; Saccharomyces cerevisiae ; 2 μm circle
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Until recently it was assumed that any short segment of DNA could be approximated as a straight rod. Many instances, however, have been reported in which the helical axis is curved. We have devised a simple method for selective identification of DNA segments containing a sequence-directed bend (curvature), by means of a two-dimensional polyacrylamide gel electrophoresis. In order to gain general insights into the structural features and the functional significance of sequence-directed bends, a bank of plasmids carrying bent DNA inserts from the Saccharomyces cerevisiae total genomic DNA was constructed. Primary characterizations of a set of bent DNA segments randomly cloned from S. cerevisiae are presented. One of the cloned DNA segments appears to be derived from a yeast plasmid, the 2 μm circle DNA.
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  • 51
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    Molecular genetics and genomics 214 (1988), S. 85-88 
    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; α-Factor ; Cell-cycle arrest ; STE genes
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    Topics: Biology
    Notes: Summary MATa cells of Saccharomyces cerevisiae defective in both the SST1 and SST2 gene products exhibit selfarrest when they express the MFα1 gene under the control of the GAL1 promoter. This reponse to endogenously produced pheromone can be alleviated by mutations which prevent the production of, or response to, α-factor. Suppressors of the self-arrest phenotype include a class of mutants which remain responsive to low levels of pheromone, but are resistant to high levels of α-factor. One of these mutants has been mapped to chromosome X, 31 cM distal to SUP4, and defines a new locus designated STE18.
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  • 52
    ISSN: 1617-4623
    Keywords: CDC28 ; Phosphate regulation ; PHO85 ; Protein kinase ; Saccharomyces cerevisiae
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    Topics: Biology
    Notes: Summary The product of the PHO85 gene, which encodes one of the negative regulatory factors of the PHO system in Saccharomyces cerevisiae, shows significant amino acid sequence homology with the CDC28 protein kinase. However, overexpressing PHO85 did not suppress the temperature sensitive phenotype of the cdc28-1 mutation. The nucleotide sequence of the PHO85 gene strongly suggests the presence of an intron near the sequence encoding the N-terminal region.
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  • 53
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    Molecular genetics and genomics 214 (1988), S. 165-169 
    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; ARO3 gene ; DAHP synthase ; DNA sequence
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    Topics: Biology
    Notes: Summary In Saccharomyces cerevisiae, the genes ARO3 and ARO4 encode isoenzymes of 3-deoxy-D-arabino-heptulosonate-7-phosphate (DAHP) synthase. Both genes are derepressed seven-fold under the general control of amino acid biosynthesis. A previously isolated 1.7kb fragment containing the ARO3 gene and the 5′- and 3′-flanking regions was sequenced. The endpoints of the ARO3 transcript coding for a 370 amino acid protein were mapped by primer extension experiments and S1 nuclease digestion. Promoter elements involved in transcription initiation and responsible for the strong general control derepression response are discussed.
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  • 54
    ISSN: 1617-4623
    Keywords: Meiosis ; Saccharomyces cerevisiae ; Sporulation ; Inessential genes ; Meiosis
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    Notes: Summary SPR3 is one of at least nine genes which are expressed in sporulating Saccharomyces cerevisiae cells at the time of meiosis I. We show below that strains homozygous for null alleles of SPR3 are capable of normal meiosis and the production of viable ascospores. We have also monitored SPR3 expression in a series of strains that are defective in meiotic development, using an SPR3: lacZ fusion carried on a single copy plasmid. β-Galactosidase activity occurred at wild-type levels in diploid strains homozygous for mutations in spo13, rad50, rad57 and cdc9, but was greatly reduced in strains carrying cdc8 or spo7 defects. We conclude that SPR3 expression is a valid monitor of early meiotic development, even though the gene is inessential for the sporulation process.
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  • 55
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    Molecular genetics and genomics 217 (1989), S. 149-154 
    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; HOM2 gene ; Aspartic semi-aldehyde ; General control
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    Topics: Biology
    Notes: Summary In Saccharomyces cerevisiae the HOM2 gene encodes aspartic semi-aldehyde dehydrogenase (ASA DH). The synthesis of this enzyme had been shown to be derepressed by growth in the presence of high concentrations of methionine. In the present work we have cloned and sequenced the HOM2 gene and found that the promoter region of this gene bears one copy of the consensus sequence for general control of amino acid synthesis. This prompted us to study the regulation of the expression of the HOM2 gene. We have found that ASA DH is the first reported enzyme of the related threonine and methionine pathway to be regulated by the general control of amino acid synthesis.
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  • 56
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    Molecular genetics and genomics 217 (1989), S. 464-470 
    ISSN: 1617-4623
    Keywords: Nucleosome positioning ; LEU2 gene ; Saccharomyces cerevisiae ; tRNA3 Leu gene ; Chromatin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The chromatin structure of theLEU2 gene and its flanks has been studied by means of nuclease digestion, both with micrococcal nuclease and DNase I. The gene is organized in an array of positioned nucleosomes. Within the promoter region, the nucleosome positioning places the regulatory sequences, putative TATA box and upstream activator sequence outside the nucleosomal cores. The tRNA3 Leu gene possesses a characteristic structure and is protected against nucleases. Most of the 5′ flank is sensitive to DNase I digestion, although no clear hypersensitive sites were found. The chromatin structure is independent of either the transcriptional state of the gene or the chromosomal or episomal location. Finally, in the plasmid pJDB207, which lacks most of the promoter, we have found that the chromatin structure of the coding region is similar to that of the wild-type allele.
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  • 57
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    Molecular genetics and genomics 216 (1989), S. 511-516 
    ISSN: 1617-4623
    Keywords: Nonsense mutation ; Read-through ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The yeast invertase structural gene SUC2 has two naturally occurring alleles, the active one and a silent allele called suc2°. Strains carrying suc2° are unable to ferment sucrose and do not show detectable invertase activity. We have isolated suc2° and found an amber codon at position 232 of 532 amino acids. However, transformants carrying suc2° on a multicopy plasmid were able to ferment sucrose and showed detectable invertase activity. Full-length invertase was found in gels stained for active invertase and in immunoblots. Therefore we concluded that the amber codon is occasionally read as an amino acid. The calculated frequency of read-through is about 4% of all translation events.
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  • 58
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    Molecular genetics and genomics 218 (1989), S. 293-301 
    ISSN: 1617-4623
    Keywords: Sporulation ; DNA sequence ; Saccharomyces cerevisiae ; Meiosis ; Chromosome segregation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Genetic studies have previously demonstrated that the RED1 gene of Saccharomyces cerevisiae is required for chromosome segregation at the first meiotic division. Northern blot hybridization analysis indicates that the RED1 gene produces two transcripts of 2.75 and 3.2 kilobases. The major 2.75 kb transcript is not present in mitotic cells and is meiotically induced to accumulate maximally just prior to the meiosis I division. The DNA sequence of the RED1 gene was determined and used to predict the amino acid sequence of the encoded gene product. The RED1 protein is 827 amino acids in length and has a molecular weight of 95.5 kilodaltons. There is no significant homology between the RED1 amino acid sequence and other known protein sequences, including those encoded by genes essential for meiosis.
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  • 59
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    Molecular genetics and genomics 212 (1988), S. 134-141 
    ISSN: 1617-4623
    Keywords: Dihydroorotase ; URA4 ; Saccharomyces cerevisiae ; DNA sequence
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    Topics: Biology
    Notes: Summary The URA4 gene of Saccharomyces cerevisiae, coding for the third enzyme of the pyrimidine pathway, has been cloned through phenotypic complementation of a ura4 mutant of S. cerevisiae. Subcloning of an original 9 kb DNA fragment, carrying the yeast URA4 gene, allowed us to localize the gene on a 2 kb ClaI-BamHI fragment. The sequence of the URA4 structural gene and surrounding DNA was determined by the dideoxynucleotide chain termination method. The URA4 gene encodes a dihydroorotase subunit of calculated molecular weight 40600. S1 nuclease mapping indicated that transcription of URA4 is initiated at four major start sites located at positions-42,-30,-22 and-18. A set of potentially significant sequences was identified in the 5′ OH non-coding region of the gene. The deduced amino acid sequence of dihydroorotase was examined and compared with homologous amino acid sequences of Salmonella typhimurium, Escherichia coli and Drosophila melanogaster. S. cerevisiae dihydroorotase shows 40% homology with the S. typhimurium and E. coli enzymes and 23% homology with the D. melanogaster enzyme. A potential active site has been predicted for dihydroorotase from these comparisons.
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  • 60
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    Molecular genetics and genomics 211 (1988), S. 430-434 
    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; SCA gene ; RME1 gene ; Haploid meiosis ; Mating type
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    Notes: Summary We have used the special properties of the spo13-1 mutation in order to study the regulation of yeast meiosis by the mating type loci. We have found that both the rme1-1 mutation and the sca mutation allow haploid meiosis in spo13-1 strains. Therefore, haploid meiosis is regulated in the same manner as diploid meiosis. Unlike rme1-1, the sca mutation allows meiosis through derepression of the silent mating type cassettes; sca strains can sporulate only because they express both MAT a and MATα information. We have found further that sca is an allele of SIR2, one of the genes involved in repression of the silent cassettes. Therefore, the RME1 gene is the only known candidate for a master negative regulator through which the MAT locus controls meiosis.
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  • 61
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    Molecular genetics and genomics 211 (1988), S. 472-476 
    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; TRP1 promoter ; REgion of dyad symmetry ; AT rich tracts
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    Notes: Summary Transcription from the yeast TRP1 promoter results in two classes of transcript, I and II, that are influenced by different promoter elements. The 5′ flanking region contains a region of dyad symmetry (RDS) which contains a 12 nucleotide AT rich inverted repeat, separated by a 21 bp spacer region. The RDS lies within a region of the promoter required for transcription of calls II RNAs. A series of internal deletions and insertions have been constructed in vitro around the RDS and the effect of each mutation on transcription has been analysed. Deleting either of the repeats abolishes class II transcription and disruption of both repeats influenced the levels of the larger class I transcripts. Deletion of the spacer had no effect but increasing the length to 33 bp reduced transcription. These results show that the RDS is an important component of the TRP1 promoter, that both repeats must be preserved and that there is some constraint on the spacing of the repeats for maximal function.
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  • 62
    ISSN: 1617-4623
    Keywords: β-Glucosidase ; Kluyveromyces fragilis ; Saccharomyces cerevisiae ; Upstream repressing sequence ; Gene expression
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    Topics: Biology
    Notes: Summary The relationship between the promoter length of the Kluyveromyces fragilis β-glucosidase gene and the level of its expression in Saccharomyces cerevisiae was studied by gene fusion between deleted promoter fragments of various lengths and the promoterless β-galactosidase gene of Escherichia coli. The removal of a region from position-425 to-232 led to a tenfold increase in the expression of the gene. The same results were obtained for the reconstructed β-glucosidase gene with the same promoter length. It is likely that the deletion of this part of the promoter removes negative regulatory elements which are functional in Saccharomyces cerevisiae. This increase in activity is the main event which may explain the high increase in gene expression (60-fold) previously observed for an upstream deletion obtained during subcloning experiments of the β-glucosidase gene. It is also shown that the expression of the gene greatly depends upon the nature of the recipient strain, the growth phase of the cell and that of the vector carrying it.
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  • 63
    ISSN: 1617-4623
    Keywords: Aneuploidy ; Yeast ; Saccharomyces cerevisiae ; Methyl benzimidazol-2-yl carbamate ; Mitosis ; Meiosis
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    Topics: Biology
    Notes: Summary A system is described in which spontaneous and chemically-induced mitotic and meiotic hyperploidy can be assayed in the same diploid culture of Saccharomyces cerevisiae. Monitoring gene dosage changes at two loci on chromosome VIII, the test utilizes a leaky temperature-sensitive allele arg4-8 and low level copper resistance conferred by the single copy allele cup1 s. An extra chromosome VIII provides simultaneous increased dosage for both genes, resulting in colonies that are both prototrophic for arginine at 30° C and copper resistant. During mitotic cell divisions in diploids, spontaneous chromosome VIII hyperploids (trisomes and tetrasomes) occur at a frequency of 6.4×10-6 per viable cell. Among ascospores, the spontaneous chromosome VIII disome frequency is 5.5×10-6 per viable spore. The tubulin-binding reagent methyl benzimidazol-2-yl carbamate (MBC) elicits enhanced levels of mitotic and meiotic aneuploidy relative to control levels. The system represents a novel model for examining chromosome behavior during mitosis and meiosis and provides a sensitive and quantifiable procedure for examining chemically induced aneuploidy.
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  • 64
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    Molecular genetics and genomics 215 (1989), S. 401-406 
    ISSN: 1617-4623
    Keywords: Protein secretion ; Saccharomyces cerevisiae ; Invertase ; Endoplasmic reticulum ; Secretion mutants
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    Topics: Biology
    Notes: Summary Intercompartmental transport of secreted proteins in yeast was analysed using invertase mutants. Deletions and insertions at the BamHI (position +787) or the Asp718 (position +1159) sites of the SUC2 gene led to mutant proteins with different behaviour regarding secretion, localization and enzyme activity. The deletion mutants showed accumulation of core glycosylated material in the endoplasmic reticulum (ER) a decrease of secreted protein by 5%–30% and loss of enzyme activity. The secreted material was localized in the culture medium and not — as is normal for invertase-in the cell wall. No delay in transport from the Golgi to the cell surface was observed, indicating that the rate-limiting step for secretion is at the ER-Golgi stage. Two insertion mutants, pIPA and pIPB, retained enzyme activity. Mutant pIPB showed 10% secretion, while 60%–70% secretion was observed for pIPA. While the non-secreted material accumulated in the ER, the secreted material was present in the cell wall. The results suggest that the presence of structures incompatible with secretion leads to ER accumulation of mutated invertase.
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  • 65
    ISSN: 1617-4623
    Keywords: RNA splicing ; Maturase ; Recombinase ; Mitochondria ; Saccharomyces cerevisiae
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    Notes: Summary When the bI4 RNA maturase, encoded by the fourth intron of the mitochondrial cytochrome b gene of Saccharomyces cerevisiae, was expressed in Escherichia coli, formation of intra-chromosomal Lac+ recombinants was stimulated threefold. This “hyper-rec” phenotype was recA as well as recBCD dependent. The most active form of the bI4 maturase stimulated homologous recombination whereas splicing deficient mutants of bI4 maturase were either deficient in or unable to stimulate homologous recombination.
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  • 66
    ISSN: 1617-4623
    Keywords: Nucleotide sequence ; PET gene ; Mitochondrial import ; Saccharomyces cerevisiae
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    Notes: Summary Translation of mitochondrial cytochrome b mRNA in yeast is activated by the product of the nuclear gene CBS1. CBS1 encodes a 27 kDa precursor protein, which is cleaved to a 24 kDa mature protein during the import into isolated mitochondria. The sequences required for mitochondrial import reside in the amino-terminal end of the CBS1 precursor. Deletion of the 76 amino-terminal amino acids renders the protein incompetent for mitochondrial import in vitro and non-functional in vivo. When present on a high copy number plasmid and under the control of a strong yeast promoter, biological function can be restored by this truncated derivative. This observation indicates that the CBS1 protein devoid of mitochondrial targeting sequences can enter mitochondria in vivo, possibly due to a bypass of the mitochondrial import system.
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  • 67
    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; TRP4 gene ; PRtransferase ; Promoter analysis ; Regulation
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    Notes: Summary The TRP4 gene of Saccharomyces cerevisiae, which encoded anthranilate phosphoribosyl transferase (E.C.2.4.2.18), is subject to the general control of amino acid biosynthesis. The regulation takes place at the transcriptional level by increasing the amount of initiation and not by changing the stability of mRNA. We have observed a change in the utilization of TRP4 mRNA start sites, depending on whether cells were grown under repressing or derepressing conditions. The function of promoter elements has been tested by deletion analysis with a plasmid-encoded TRP4 gene. A routinely practicable method was used for copy-number calibration of plasmids based on 2 μm DNA. Promoter structures and spacing problems in the TRP4 promoter region are discussed.
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  • 68
    ISSN: 1617-4623
    Keywords: Mitochondrial introns ; Reverse transcriptase ; Saccharomyces cerevisiae
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    Notes: Summary Some pet- (or mit-) mutations impeding the splicing of one or several intron(s) of the yeast mitochondrial pre-mRNA(s) are suppressed in vivo by the DNA deletion of these introns. We have genetically demonstrated that introns aI1 and/or aI2 of the cytochrome c oxidase subunit 1 gene are necessary for this deletion process. The facts that adjacent introns are simultaneously deleted and that, in the pet- (or mit-) mutants which easily revert by intron deletion, the splicing of the introns they affect is only partially blocked, suggest that the intron encoded proteins aI1 and/or aI2 could intervene by means of their putative reverse transcriptase activity.
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  • 69
    ISSN: 1615-6102
    Keywords: Microtubule neoformation ; Nocodazole ; Protoplasts ; Saccharomyces cerevisiae
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    Topics: Biology
    Notes: Summary By following microtubule neoformation after their complete destruction by nocodazole, we analyzed the pattern of microtubule nucleation in protoplasts ofSaccharomyces cerevisiae. Using immunofluorescence, the drug was shown to induce rapid and complete disassembly of both cytoplasmic and spindle microtubules and to selectively block protoplast nuclear division at a defined stage of the cell cycle. Treated protoplasts placed in a drug-free environment recovered a more abundant microtubular system. The majority of microtubules re-formed at SPBs whereas a minority of free-ended microtubules nucleated in the cytoplasm of the protoplasts without any detectable association with recognizable nucleation sites. Random nucleation of free microtubules might be induced by high amounts of unpolymerized tubulin likely to be present in the protoplasts at the moment of drug release.
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  • 70
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    Aquatic ecology 22 (1988), S. 31-34 
    ISSN: 1573-5125
    Keywords: estuaries ; tidal mixing ; suspended matter ; sedimentation ; nitrogen ; plankton ; benthos
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract An estuary such as the Westerschelde is a highly dynamic environment, both on an ecological time scale where climatic and hydrodynamic forces, mainly the tides, shape a very variable environment and on a geological, evolutionary time scale, since estuaries are young and very unstable habitats. Low species diversity and high adaptability of the resident animal and plant populations are characteristic of estuarine habitats where large fluctuations in submersion, salinity, temperature etc. occur. The existing biota are therefore resilient to environmental stress and effects of the important influx of anorganic and organic pollutants from the river Schelde and its tributaries on the biota in the estuary are not easy to detect. Although water movement and sedimentation patterns in the Westerschelde are relatively well known, there exists little information on important ecological processes such as primary production and heterotrophic metabolism in the estuary.
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  • 71
    ISSN: 1573-5036
    Keywords: carbon ; erosion ; land clearing ; nitrogen ; noncalcic brown soils ; phosphorus ; spatial variability ; xerosols
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Several transects of topsoil samples were taken immediately after land clearing and one year later from a savannah soil in the semiarid NE of Brazil. Natural spatial variability of key fertility indicators (C, N, P) was large with coefficients of variation 〉50%. This variability was related to heterogeneity of the soil parent material, and to relative slope position which affected deposition and removal of erodible materials. The distribution of gravel and different particle size fractions was an indicator of the variability as related to soil formation and erosional resorting. One year after the site was cleared and planted to trees, the decrease in C, N and resin-extractable P was in the same range as the initial spatial variability. Treatment effects were therefore difficult to observe but became more obvious when regression analysis on soil textural components was used to reduce data variability.
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  • 72
    ISSN: 1573-5036
    Keywords: actinorhizal ; Ceanothus ; Frankia ; nitrogen ; propagation ; inoculation ; nodulation
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    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Rooted cuttings ofCeanothus griseus varhorizontalis were irrigated with 0, 10, 20, 50, 75 or 100ppm nitrogen as NH4NO3 for eight weeks prior to inoculation with infectiveFrankia. After inoculation, half of the plants for each treatment nitrogen level continued to be irrigated with the preconditioning nitrogen level and half were given no more supplemental nitrogen. For plants continuously receiving nitrogen, nodule initiation (nodule number) was inversely correlated with increasing supplemental nitrogen levels, and suppressed above 50 ppm N. Leaf nitrogen above 2% in continuous-N plants correlated with greatly reduced or suppressed nodulation. Plants maintained after inoculation without supplemental nitrogen showed influence of the prior nitrogen treatment on nodulation. Preconditioning at 50 ppm and above greatly reduced the number of nodules formed. The evidence suggests that stored internal nitrogen can regulate nodulation. Plant biomass accumulated maximally when nodulation was suppressed, at 75 and 100 ppm supplemental N applied continuously. Internode elongation during the nodulation period occurred only on nodulated plants, or in the presence of supplemental N (10 ppm and above).
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  • 73
    ISSN: 1573-9104
    Keywords: barley ; Triticeae ; grain ; amino acid composition ; nitrogen ; chemical score
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Barley grains (9 samples from 7 cultivars) with nitrogen contents (N) ranging from 1.45 to 4.01% of dry matter were analysed for their amino acid (AA) composition with high accuracy from six different hydrolysates per sample. AA levels in grain increased as linear functions ofN with correlation coefficients close to unity. A comparison with literature data confirmed that the AA composition of any grain sample of normal barley can be predicted from itsN for all phenotypes and genotypes. AAs in grain protein changed as hyperbolic functions ofN which increased for Phe, Pro and Glx but more or less strongly decreased for the other AAs. By plotting AA scores againstN, barley proteins were shown to be always richer than wheat and rye in Val and Phe + Tyr; sometimes richer than both other species forN〈2 (Lys); 2.2 (Leu and Ile); 3.4 (Thr); sometimes intermediate to wheat and rye above the latterN values. They were also intermediate in sulphur AAs forN〈1.9 and drastically poorer forN〉1.9. However, they were richer than both other species in Trp forN〉1.6. The hyperbolic variations of non-protein nitrogen and nitrogen-to-protein conversion factors were determined as a function ofN and also compared with those of wheat and rye.
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    Molecular genetics and genomics 218 (1989), S. 531-535 
    ISSN: 1617-4623
    Keywords: Origin of replication ; Promoter ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In an attempt to devise a new assay for ARS-binding proteins we have inserted the HO ARS between the upstream activation site and the TATA region of the yeast CYC1 promoter. A marked reduction in promoter activity is observed. Inactivation of the HO ARS element by point mutation does not restore promoter activity to its original level, although a modest activation is seen. We have also inserted the HO ARS into the intron of the yeast actin gene; although there is no apparent deleterious effect on transcription, the activity of the ARS is abolished in this new environment.
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  • 75
    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; Heavy metal resistance ; DNA sequence ; Membrane
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    Notes: Summary A DNA fragment conferring resistance to zinc and cadmium ions in the yeast Saccharomyces cerevisiae was isolated from a library of yeast genomic DNA. Its nucleotide sequence revealed the presence of a single open reading frame (ORF; 1326 bp) having the potential to encode a protein of 442 amino acid residues (molecular mass of 48.3 kDa). A frameshift mutation introduced within the ORF abolished resistance to heavy metal ions, indicating the ORF is required for resistance. Therefore, we termed it the ZRC1 (zinc resistance conferring) gene. The deduced amino acid sequence of the gene product predicts a rather hydrophobic protein with six possible membrane-spanning regions. While multiple copies of the ZRC1 gene enable yeast cells to grow in the presence of 40 mM Zn2+, a level at which wild-type cells cannot survive, the disruption of the chromosomal ZRC1 locus, though not a lethal event, makes cells more sensitive to zinc ions than are wild-type cells.
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  • 76
    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; a-factor ; Conjugation ; G1 arrest ; ssl mutations
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Nine independent mutants which are supersensitive (ssl −) to G1 arrest by the mating hormone a-factor were isolated by screening mutagenized Saccharomyces cerevisiae MATα cells on solid medium for increased growth inhibition with a-factor. These mutants carried lesions in two complementation groups, ssl1 and ssl2. Mutations at the ssl1 locus were mating type specific: MATα ssl1 − cells were supersensitive to α-factor but MATα ssl1 − were not supersensitive to α-factor. In contrast, mutations at the ssl2. locus conferred supersensitivity to the mating hormone of the opposite mating type on both MATα, and MATa cells. The α-cell specific capacity to inactivate externally added a-factor was shown to be lacking in MATα ssl1 − mutants whereas MATα ssl2. cells were able to inactivate a-factor. Complementation analysis showed that ssl2 and sst2, a mutation originally isolated as conferring supersensitivity to α-factor to MATa cells, are lesions in the same gene. The ssl1 gene was mapped 30.5 centi-Morgans distal to ilv5 on chromosome XII.
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  • 77
    ISSN: 1617-4623
    Keywords: Aspergillus oryzae ; Alkaline protease ; Prepro sequence ; Heterologous expression ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have cloned and determined the nucleotide sequence of a cDNA fragment for the entire coding region of the alkaline protease (Alp) from a filamentous ascomycete Aspergillus oryzae. According to the deduced amino acid sequence, Alp has a putative prepro region of 121 amino acids preceding the mature region, which consists of 282 amino acids. A consensus sequence of a signal peptide consiting of 21 amino acids is found at the N-terminus of the prepro region. The primary structure of the mature region shares extensive homology (29%–44%) with those of subtilisin families, and the three residues (Asp 32, His 64 and Ser 221 in subtilisin BPN′) composing the active site are preserved. The entire cDNA, coding for prepro Alp, when introduced into the yeast Saccharomyces cerevisiae, directed the secretion of enzymatically active Alp into the culture medium, with its N-terminus and specific activity identical to native Aspergillus Alp.
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  • 78
    ISSN: 1617-4623
    Keywords: Gene conversion ; Crossing-over ; Mismatch repair ; Saccharomyces cerevisiae ; Psoralens
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The induction of gene conversion and mitotic crossing-over by photoaddition of psoralens, 254 nm ultraviolet radiation, and nitrogen mustards was determined in diploid cells homozygous for the pso3-1 mutation and in the corresponding wild type of Saccharomyces cerevisiae. For these different agents, the frequency of non-reciprocal events (conversion) is reduced in the pso3-1 mutant compared to the wild type. In contrast, the frequency of reciprocal events (crossing-over) is increased at a range of doses. These observations, together with the block in induced mutagenesis for both reverse and forward mutations previously reported for the pso3-1 mutant, suggest that the PS03 gene product plays a role in mismatch repair of short patch regions. The block in gene conversion in the pso3 homozygous diploid leads, in the case of nitrogen mustards, to specific repair intermediates which are lethal to the cells.
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  • 79
    ISSN: 1617-4623
    Keywords: Enhanced secretion ; Human lysozyme production ; Protease mutant ; Protein processing ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Yeast mutant strains which secrete large amounts of human lysozyme were screened using an agar medium containing bacterial cells. Nine mutants secreted over 10 times more lysozyme than the wild-type parent strain. The mRNA levels for lysozyme in the mutants were not higher than that of the wild-type strain. Three of the mutant strains were deficient in carboxypeptidase Y activity. It was found that the protease deficiency was caused by a deficiency in conversion of proenzyme to mature enzyme in ssl1 mutant cells. The ssl1 gene was found to be closely linked to the centromere and determine both the efficiency of secretion of lysozyme and the processing of carboxypeptidase Y.
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  • 80
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    Molecular genetics and genomics 219 (1989), S. 495-498 
    ISSN: 1617-4623
    Keywords: ARS1 ; Plasmid multimerization ; RAD52 ; Saccharomyces cerevisiae ; Eukaryotic recombination
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A mutant plasmid, pX, derived from the 1453 base pair small plasmid, YARp1 (or TRP1 RI circle), consists of 849 base pairs of DNA bearing the TRP1 gene and the ARS1 sequence of Saccharomyces cerevisiae and, unlike YARp1 and other commonly used yeast plasmids, highly multimerizes in a S. cerevisiae host. The multimerization of pX was dependent on RAD52, which is known to be necessary for homologous recombination in S. cerevisiae. Based upon this observation, a regulated system of multimerization of pX with GAL1 promoter-driven RAD52 has been developed. We conclude that the regulated multimerization of pX could provide a useful model system to study genetic recombination in the eukaryotic cell, in particular to investigate recombination intermediates and the effects of various trans-acting mutations on the multimerization and recombination of plasmids.
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  • 81
    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; Yeast ; Copy number ; Thymidine kinase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A 2 μm circle-based chimaeric plasmid containing the yeast LEU2 and the Herpes Simplex Virus type 1 thymidine kinase (HSV-1 TK) genes was constructed. Transformants grown under selective conditions for the LEU2 gene harboured the plasmid at about 15 copies per cell whilst selection for the HSV-1 TK gene led to an increase to about 100 copies per cell. Furthermore, the plasmid copy number could be controlled by the stringency of selection for the TK gene, and the increase in TK gene dosage was reflected in an increase in intracellular thymidine kinase activity. The mitotic stability of the plasmid in “high-copy” and “low-copy” number cells was determined. “High-copy” number cells showed a greater mitotic stability. The relationship of TK expression to plasmid copy number may be useful for the isolation of plasmid copy number mutants in yeast and the control of heterologous gene expression.
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  • 82
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    Molecular genetics and genomics 212 (1988), S. 426-431 
    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; 2 μm circle ; Plasmid-partitioning
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The yeast 2 μm circle encodes four major transcribed open reading frames, A, B, C and D. Products of ORF's A, B and C, together with the inverted repeats and the other cis-acting loci ORI and STB, have been shown to be involved in plasmid maintenance. However, the function of ORF D has remained unclear. We have therefore carried out studies on 2 μm derivatives with both insertional and frameshift mutations in D. Our results indicate that there is a protein product encoded by ORF D, which is involved in plasmid maintenance. When the copy number of the C gene was reduced to one, by chromosomal integration, we observed striking differences in the efficiency of partitioning of D + and D − plasmid derivatives. Absence of D function could be compensated by an increase in dosage of the C gene, indicating that the D product may act to regulate C expression. Since the C product has been implicated in copy number control as well as partitioning, our data suggest that the D product may also be involved in both of these processes.
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  • 83
    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; TRP1 gene ; Yeast promoter ; Yeast vectors ; Copy number
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The TRP1 gene of Saccharomyces cerevisiae is the only TRP gene which is not derepressible by the general control regulatory system. In the TRP1 promoter transcription starts at five initiation sites, organized in two clusters. The two transcripts of the first, more upstream cluster include a long leader sequence of approximately 200 bp. A transcriptional terminator element located in the 5′ region of the TRP1 gene is essential for accurate gene expression. In partial TRP1 promoters lacking the terminator, like the original EcoRI TRP1 fragment used in numberous vectors, plasmid-encoded transcription is initiated predominantly in adjacent vector regions, resulting mainly in large, poorly translated transcripts. This poor translation is not due to mRNA instability. The effect can be suppressed by introducing artificial transcription barriers between vector sequences and the truncated EcoRI TRP1 fragment.
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  • 84
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    Molecular genetics and genomics 213 (1988), S. 425-434 
    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; Mitochondria ; Transfer RNA ; syn - mutation ; Revertants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We isolated revertants capable of respiration from the respiratory deficient yeast mutant, FF1210-6C/ 170, which displays greatly decreased mitochondrial protein synthesis due to a single base substitution at the penultimate base of the tRNAAsp gene on mitochondrial (mt) DNA. Three classical types of revertant were identified: (1) same-site revertants; (2) intragenic revertants which restore the base pairing in the acceptor stem of the mitochondrial tRNAAsp; and (3) extragenic suppressors located in nuclear DNA. In addition a fourth type of revertant was identified in which the mutant tRNAAsp is amplified due to the maintenance of both the original mutant mtDNA and a modified form of the mutant mtDNA in which only a small region around the tRNAAsp gene is retained and amplified. The latter form resembles the mtDNA in vegetative petite (rho -) strains which normally segregates rapidly from the wild-type mtDNA. Each revertant type was characterized genetically and by both DNA sequence analysis of the mitochondrial tRNAAsp gene and analysis of the quantity and size of RNA containing the tRNAAsp sequence. These results indicate that the mitochondrial tRNAAsp of the mutant retains a low level of activity and that the presence of the terminal base pair in tRNAAsp is a determinant of both tRNAAsp function and the maintenance of wild-type levels of tRNAAsp.
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  • 85
    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; URA3 locus ; Cyclobutane dimer ; Pyrimidine-pyrimidone (6-4) photoproduct ; “A rule”
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A forward mutation system has been developed to obtain rapidly clonable mutants at the URA3 locus in yeast by means of selection for 5-fluoroorotic acid resistance. We have used this system to determine base changes in 35 spontaneous and 34 ultraviolet radiation-induced ura3 base substitution mutants. Other mutants (frameshift, deletion, duplication, replacement) were detected as well. Evidence is reported which suggests cyclobutane dimers are the principal mutagenic lesions induced by UV radiation in stationary phase cells of the yeast Saccharomyces cerevisiae. Since most of the induced lesions are at 5′-TT-3′ sites, the results suggest that the “A-rule”, preferential insertion of adenine residues opposite poorly pairing sites in DNA, does not apply for yeast cells irradiated in stationary phase, whereas the spontaneous mutation data indicate that the A-rule applies for cells in logarithmic phase. Most of the spontaneous mutations are transversions. UV-induced transitions and transversions occur at approximately equal frequencies.
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  • 86
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    Molecular genetics and genomics 214 (1988), S. 425-432 
    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; Mitochondrial DNA ; Intergenic sequences ; Transmission ; Genetic crosses
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Mitochondrial DNAs (mtDNA) from four stable revertant strains generated from high frequency petite forming strains of Saccharomyces cerevisiae have been shown to contain deletions which have eliminated intergenic sequences encompassing ori1, ori2 and ori7. The deleted sequences are dispensable for expression of the respiratory phenotype and mutant strains exhibit the same relative amount of mtDNA per cell as the wild-type (wt) parental strain. These deletion mutants were also used to study the influence of particular intergenic sequences on the transmission of closely linked mitochondrial loci. When the mutant strains were crossed with the parental wt strains, there was a strong bias towards the transmission into the progeny of mitochondrial genomes lacking the intergenic deletions. The deficiency in the transmission of the mutant regions was not a simple function of deletion length and varied between different loci. In crosses between mutant strains which had non-overlapping deletions, wt mtDNA molecules were formed by recombination. The wt recombinants were present at high frequencies among the progeny of such crosses, but recombinants containing both deletions were not detected at all. The results indicate that mitochondrial genomes can be selectively transmitted to progeny and that two particular intergenic regions positively influence transmission. Within these regions other sequences in addition to ori/rep affect transmission.
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  • 87
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    Molecular genetics and genomics 214 (1988), S. 263-270 
    ISSN: 1617-4623
    Keywords: DNA sequence ; PET gene ; Saccharomyces cerevisiae ; Transcription initiation ; Translation activator
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In yeast, synthesis of apocytochrome b from mitochondrial COB mRNA depends on at least three nuclear gene products. The translation stimulatory effect by two of these nuclear genes, CBS1 and CBS2, is mediated by the 5′-untranslated leader of COB mRNA. In this report, we show that CBS2 is located on chromosome IV and provide genetic evidence that the CBS2 gene encodes a polypeptide. Determination of the DNA sequence reveals a contiguous open reading frame of 1167 bp. The deduced polypeptide has a calculated molecular weight of 44.5 kDa and is characterized by a high content of positively charged amino acids. It has no significant homology to any known protein. The CBS2 gene is transcribed into low abundance mRNA species with a major transcription initiation site located 97 bp upstream from the ATG start codon next to a poly(dA-dT) stretch.
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  • 88
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    Molecular genetics and genomics 215 (1989), S. 455-462 
    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; Orotate phosphoribosyl transferase ; Nucleotide sequence ; Transcription ; Regulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The URA5 gene of Saccharomyces cerevisiae encodes orotate phosphoribosyl transferase (EC 2.4.2.10; OPRTase) which catalyses the transformation of orotate to OMP in the pyrimidine pathway. We present in this paper the cloning and the sequencing of this gene, the last in the yeast pyrimidine pathway to be cloned. We have deduced the protein sequence of the OPRTase of S. cerevisiae from the DNA sequence and compared it to that of Escherichia coli, Podospora anserina and Dictyostelium discoideum. Some important similarities in the structure of these four proteins have been found. Finally, we have quantified the transcription of the URA5 gene in different physiological conditions and confirmed that it was not under the control of UTP or any intermediary product of the pathway.
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  • 89
    ISSN: 1617-4623
    Keywords: DNA sequence ; PET gene ; Saccharomyces cerevisiae ; Mitochondrial import ; cytochrome c oxidase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The yeast nuclear SCO1 gene is required for accumulation of the mitochondrially synthesized cytochrome c oxidase subunits I and II (COXI and COXII). We cloned and characterized the SCO1 gene. It codes for a 0.9 kb transcript. DNA sequence analysis predicts a 33 kDa protein. As shown by in vitro transcription and translation experiments in combination with import studies on isolated mitochodria, this protein is matured into a 30 kDa polypeptide which is tightly associated with a mitochondrial membrane. The possible function of the SCO1 gene product in the assembly of cytochrome c oxidase is discussed.
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  • 90
    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; GLN1 ; Glutamine synthetase ; Regulatory systems ; Transcription
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary TheGLN1 gene ofSaccharomyces cerivisiae was cloned by complementation of agln1 auxotroph. AGLN1-lacZ fusion was constructed to assayGLN1 promoter activity. β-Galactosidase and glutamine synthetase expression in chromosomally integratedGLN1-lacZ fusion strains were co-regulated in response to a shift from glutamine to glutamate as the nitrogen source, purine limitation, and 3-aminotriazole-induced histidine starvation. Regulation ofGLN1 expression by each of the three pathways occurred at the transcriptional level. Increased accumulation ofGLN1 mRNA was observed within 5 min after a shift from glutamine to glutamate as the nitrogen source. After 5 min following glutamine addition to the cells growing with glutamate as nitrogen source. This indicates that theGLN1 message is unstable and has a half-life of approximately 3 min. Deletion analysis indicated that the sequences required forGLN1 expression are located within approximately 350 bp upstream from the transcriptional initiation site.
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  • 91
    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; DNA repair ; Cross-link ; Transposon mapping ; Nitrogen mustard
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have isolated yeast gene SNM1 via complementation of sensitivity towards bi- and tri-functional alkylating agents in haploid and diploid yeast DNA repair-deficient snm1-1 mutants. Four independent clones of plasmid DNA containing the SNM1 locus were isolated after transformation with a YEp24-based yeast gene bank. Subcloned SNM1-containing DNA showed (i) complementation of the repair-deficiency phenotype caused by either one of the two different mutant alleles snm1-1 and snm1-2 ts; (ii) complementation in haploid and diploid yeast snm1-1 mutants by either single or multiple copies of the SNM1 locus; and (iii) that the SNM1 gene is at most 2.4 kb in size. Expression of SNM1 on the smallest subclone, however, was under the control of the GAL1 promotor. Gene size and direction of transcription was further verified by mutagenesis of SNM1 by Tn10-LUK transposon insertion. Five plasmids containing Tn10-LUK insertions at different sites of the SNM1-containing DNA were able to disrupt function of genomic SNM1 after gene transplacement. Correct integration of the disrupted SNM1::Tn10-LUK at the genomic site of SNM1 was verified via tetrad analysis of the sporulated diploid obtained after mating of the SNM1::Tn10-LUK transformant to a haploid strain containing the URA3 SNM1 wild-type alleles. The size of the poly(A)+ RNA transcript of the SNM1 gene is 1.1 kb as determined by Northern analysis.
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  • 92
    ISSN: 1617-4623
    Keywords: pso4-1 mutation ; Saccharomyces cerevisiae ; Error-prone recombinational repair ; Mitotic recombination
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The induction of mitotic gene conversion and crossing-over inSaccharomyces cerevisiae diploid cells homozygous for thepso4-1 mutation was examined in comparison to the corresponding wild-type strain. Thepso4-1 mutant strain was found to be completely blocked in mitotic recombination induced by photoaddition of mono- and bifunctional psoralen derivatives as well as by mono- (HN1) and bifunctional (HN2) nitrogen mustards or 254 nm UV radiation in both stationary and exponential phases of growth. Concerning the lethal effect, diploids homozygous for thepso4-1 mutation are more sensitive to all agents tested in any growth phase. However, this effect is more pronounced in the G2 phase of the cell cycle. These results imply that the ploidy effect and the resistance of budding cells are under the control of thePSO4 gene. On the other hand, thepso4-1 mutant is mutationally defective for all agents used. Therefore, thepso4-1 mutant has a generalized block in both recombination and mutation ability. This indicates that thePSO4 gene is involved in an error-prone repair pathway which relies on a recombinational mechanism, strongly suggesting an analogy between thepso4-1 mutation and theRecA orLexA mutation ofEscherichia coli.
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  • 93
    ISSN: 1617-4623
    Keywords: GC clusters ; Mobile elements ; Target sites ; mtDNA ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary GC clusters constitute the major repetitive elements in the mitochondrial (mt) genome of the yeast Saccharomyces cerevisiae. Many of these clusters are optional and thus contribute much to the polymorphism of yeast mtDNAs. We have made a systematic search for polymorphic sites by comparing mtDNA sequences of various yeast strains. Most of the 26 di- or polymorphic sites found differ by the presence or absence of a GC cluster of the majority class, here referred to as the M class, which terminate with an AGGAG motif. Comparison of sequences with and without the GC clusters reveal that elements of the subclasses M1 and M2 are inserted 3′ to a TAG, flanked by A+T rich sequences. M3 elements, in contrast, only occur in tandem arrays of two to four GC clusters; they are consistently inserted 3′ to the AGGAG terminal sequence of a preexisting cluster. The TAG or the terminal AGGAG, therefore, are regarded as being part of the target sites for M1 and M2 or M3 elements, respectively. The dinucleotide AG is in common to both target sites; it also occurs at the 3′ terminus (AGGAG). This suggests its duplication during GC cluster insertion. This notion is supported by the observation that GC clusters of the minor classes G and V similarily repeat at their 3′ terminus a GT or an AA dinucleotide, respectively, from their putative target sites.
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  • 94
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    Molecular genetics and genomics 218 (1989), S. 57-63 
    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; PET gene ; Transcriptional regulation ; Anaerobiosis ; 5′ mapping
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Translation of mitochondrial cytochrome b RNA in yeast requires the product of the nuclear gene CBS1, a 27.5 kDa soluble mitochondrial protein. In this paper we show that the CBS1 gene is located on chromosome IV immediately adjacent to COX9, the gene coding for cytochrome c oxidase subunit VIIa. CBS1 is transcribed as a very low abundant 900 b RNA. Transcription starts at a single position 101 bp upstream of the CBS1 initiation codon. At positions-39 to-27 of its leader sequence it contains a small open reading frame of 4 codons. By monitoring the β-galactosidase activity of a CBS1/lacZ fusion construct we show that expression of CBS1 is subjected to regulation by oxygen and by glucose: the β-galactosidase activity is elevated threefold in glycerol or galactose grown cells compared to that in glucose grown cells. A further threefold reduction of the activity is observed in anaerobically grown cells. In accordance with this result is the observation that the steady-state level of CBS1 mRNA of anaerobically grown cells is ninefold lower than that of aerobically cultured cells.
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  • 95
    ISSN: 1573-5036
    Keywords: added nitrogen interaction ; green manure ; 15N ; N loss ; neem ; nitrate ; nitrogen ; Oryza sativa L. ; Vigna unguiculata (L.) Walp
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Little is known about whether the high N losses from inorganic N fertilizers applied to lowland rice (Oryza sativa L.) are affected by the combined use of either legume green manure or residue with N fertilizers. Field experiments were conducted in 1986 and 1987 on an Andaqueptic Haplaquoll in the Philippines to determine the effect of cowpea [Vigna unguiculata (L.) Walp.] cropping systems before rice on the fate and use efficiency of15N-labeled, urea and neem cake (Azadirachta indica Juss.) coated urea (NCU) applied to the subsequent transplanted lowland rice crop. The pre-rice cropping systems were fallow, cowpea incorporated at the flowering stage as a green manure, and cowpea grown to maturity with subsequent incorporation of residue remaining after grain and pod removal. The incorporated green manure contained 70 and 67 kg N ha−1 in 1986 and 1987, respectively. The incorporated residue contained 54 and 49 kg N ha−1 in 1986 and 1987, respectively. The unrecovered15N in the15N balances for 58 kg N ha−1 applied as urea or NCU ranged from 23 to 34% but was not affected by pre-rice cropping system. The partial pressure of ammoniapNH3, and floodwater (nitrate + nitrite)-N following application of 29 kg N ha−1 as urea or NCU to 0.05-m-deep floodwater at 14 days after transplanting was not affected by pre-rice cropping system. In plots not fertilized with urea or NCU, green manure contributed an extra 12 and 26 kg N ha−1, to mature rice plants in 1986 and 1987, respectively. The corresponding contributions from residue were 19 and 23 kg N ha−1, respectively. Coating urea with 0.2g neem cake per g urea had no effect on loss of urea-N in either year; however, it significantly increased grain yield (0.4 Mg ha−1) and total plant N (11 kg ha−1) in 1987 but not in 1986.
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  • 96
    ISSN: 1573-5036
    Keywords: decomposition ; green manure ; litterbag ; nitrogen ; paddy rice ; Sesbania sesban ; Sri Lanka
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Sesbania sesban was evaluated as green manure crop for lowland rice in the Dry Zone of Sri Lanka. The legume was grown during a fallow period before lowland rice (Oryza sativa) and ploughed under just before transplanting. Weight loss and nitrogen content in litterbags containing leaves, stems and roots of the legume were monitored. Comparisons were made between rice yields from 20 m2 plots after green manuring in combination with different nitrogen fertilizer levels (0, 2.4, 4.8 and 7.2 gm−2) and nitrogen fertilizer (9.6 gm−2) alone. Above-ground biomass ofS. sesban was 440 gm−2 (dry wt) when ploughed under after 84 days growth. N-content in leaves, stems and roots was 3.76%, 0.41% and 0.73%, respectively. This gave a N-input fromS. sesban of 9.2 gm−2 (8.3 g from above-ground parts and 0.9 g from roots). The corresponding K and P inputs were 7.3 and 0.6 gm−2 respectively. The nitrogen rich leaves, which contained 88% of the nitrogen in the above-ground parts, decomposed and released its nitrogen much more rapidly than the stems and roots. After only four days the leaves had released 5.3 g Nm−2 and after 14 days they had released 6.4 g Nm−2. The highest rice yield (505 gm−2) was obtained usingS. sesban and 4.8 gm−2 of N-fertilizer. The yields with only N-fertilizer or onlyS. sesban were 442 gm−2 and 396 gm−2, respectively. Due to the rapid decomposition of the nitrogen rich leaves,S. sesban did not behave as a slow release fertilizer. Thus, it is not necessary to apply nitrogen fertilizers as a basal dose.
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  • 97
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    Plant and soil 116 (1989), S. 77-84 
    ISSN: 1573-5036
    Keywords: alfalfa ; amino acids ; ammonium ; Glycine max (L.) Merr. ; leachates ; lucerne ; Medicago sativa L. ; nitrogen ; rhizosphere ; root exudates ; soyabean
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract An enclosed root chamber containing sterile sand medium was used to study net nitrogen (N) release from actively growing root systems of ‘Saranac’ alfalfa (Medicago sativa L.) and ‘Fiskeby V’ soybean (Glycine max L. Merr.). Plants were inoculated with a rhizobial strain appropriate to each host, irrigated with N-free nutrient solution, and grown either to 85 or to 173 d after germination (alfalfa) or to physiological maturity (soybean). Alfalfa released 4.5% of symbiotically-fixed plant N into the root zone over its growth period; soybean released 10.4% of plant N. Root zone leachates were analyzed for total N and for amino acid and ammonium content. Significant ammonium-N release occurred from the alfalfa but not the soybean root system; little amino-N was released by root systems of either species. Shoot harvest and water deficit caused increased release of N from alfalfa roots. The results provide evidence that alfalfa and soybean released significant proportions of their N into the root zone, and indicate that while substantial ammonium-N was released from alfalfa roots, passive leakage of amino-N was not a primary mechanism for N release from root systems of either species.
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  • 98
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    Plant and soil 106 (1988), S. 35-41 
    ISSN: 1573-5036
    Keywords: carbon ; crop residue ; mineralization ; nitrogen ; organic matter ; sulfur
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The mineralization of C, N, and S from residues of three different crop species (wheat, lentil, and rape) grown under diverse nutritional regimes was measured over a 12-week incubation period under controlled conditions. The rate of decomposition, as measured by CO2 evolution, varied considerably among treatments and appeared to be controlled almost entirely by N content of the residue (R2=0.98). Similarly, N mineralization was strongly tied to N concentration. The critical N concentration, below which significant immobilization of N occurred, declined over time, ranging from 1.9% at day 14 to 1.1% at day 84. Mineralization of S was positively correlated with initial S concentration (R2=0.95) and negatively related to N concentration, apparently because of a dilution effect. The results demonstrate that decomposition and N and S mineralization of crop residues, under conditions prevalent in the experiment, are primarily a function of their nutrient concentrations rather than biochemial composition related to crop species. As a result, it should be possible to enhance rate of residue decomposition, increase quantities of N and S mineralized, and avert detrimental immobilization losses in the following year by governing the nutritional regime under which the crop is grown.
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  • 99
    ISSN: 1573-5036
    Keywords: decomposition ; lignin ; litter ; nitrogen ; sub-tropical-forest
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The effect of the initial lignin and nitrogen contents of the leaves of five tree species, on their decomposition dynamics, was studied in the sub-tropical forest ecosystem existing at Shiroy hills. The decomposition dynamics of litter materials are described by inverse linear relationships between the percentage of the original biomass remaining and the nitrogen concentration in the residual material. Initial lignin and nitrogen were highly correlated with remaining biomass (r=0.94 and 0.77). The rate constants (K), for the annual leaf mass loss, ranged from −0.18 to −0.56. The values for initial lignin (%), initial nitrogen (%), the ratio between initial lignin, initial nitrogen and the annual decomposition rate constants were compared with similar results from other studies in various climatic zones of the world.
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  • 100
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    Plant and soil 105 (1988), S. 105-111 
    ISSN: 1573-5036
    Keywords: chronosequence ; desert ; ion-exchange resins ; mineralization ; nitrogen ; nutrient availability ; phosphorus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Anin situ resin bag technique was used to measure the relative availabilities of N and P along a chronosequence of soils in southern New Mexico, and was compared to two more common indices of nutrient availability. Accumulations of N and P during 10-week intervals over an 18 month period were separable into wet season (September–January) and dry season (February–August) groups, with wet season values significantly greater than dry season values. Only accumulations during the wet season showed significant differences among sites, thus stressing the role of field water regime in interpreting resin accumulation results. Total mineral N (NO3+NH4) sorbed by resins was significantly correlated to laboratory N mineralization rates. Although accumulation patterns of N and P were similar to patterns of %N and %P in shrub species growing along the chronosequence, these similarities were not statistically significant. A laboratory experiment demonstrated that bicarbonate-form anion resins are preferable to hydroxyl-form resins, as long as standards are made from solutions extracted by resins to account for variable ion recovery efficiencies.
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