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  • Articles  (3,484)
  • Nature Publishing Group  (2,513)
  • Wiley-Blackwell  (793)
  • Blackwell Publishing Ltd  (178)
  • 1995-1999  (3,484)
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  • 1995-1999  (3,484)
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  • 1
    Electronic Resource
    Electronic Resource
    Energy requirement for pullulanase secretion by the main terminal branch of the general secretory pathway (1997)
    Oxford BSL : Blackwell Publishing Ltd
    Molecular microbiology 24 (1997), S. 0 
    Details
    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: The energy requirement for the second step in pullulanase secretion by the general secretory pathway was studied in Escherichia coli. In order to uncouple the two steps in the secretion pathway (across the cytoplasmic and outer membranes, respectively) and to facilitate kinetic analysis of secretion, a variant form of pullulanase lacking its N-terminal fatty acid membrane anchor was used. The transport of the periplasmic secretion intermediate form of this protein across the outer membrane was not inhibited by concentrations of sodium arsenate in excess of those required to reduce ATP levels to ≤10% of their normal value. Pullulanase secretion was inhibited by the protonophore carbonyl cyanide m-chlorophenyl hydrazone at concentrations which were similar to those reported by others to be required to prevent solute uptake or the export and processing of preproteins across the cytoplasmic membrane, but which were in excess of those required to fully dissipate the proton-motive force and to reduce lactose uptake to a significant extent.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1365-2958.1997.3451726.x
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  • 2
    Electronic Resource
    Electronic Resource
    Transcriptional regulation of the Yersinia pseudotuberculosis pH 6 antigen adhesin by two envelope-associated components (1997)
    Oxford BSL : Blackwell Publishing Ltd
    Molecular microbiology 24 (1997), S. 0 
    Details
    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: The Yersinia pseudotuberculosis pH 6 antigen mediates haemagglutination and adhesion to cultured mammalian cells. The synthesis of pH 6 antigen requires the products of the psaEFABC genes in both Yersinia pseudotuberculosis and Escherichia coli. In-frame deletion mutations of psaE and psaF caused defective haemagglutination. In contrast, we showed that the psaABC genes were sufficient for haemagglutination if they were expressed by a heterologous promoter. Environmental regulation of pH 6 antigen by temperature and pH occurs via regulation of the major pilus protein PsaA at the transcriptional level. Northern blot analyses indicate that the psaA transcript was absent in either psaE or psaF mutant strains. Primer extension analyses indicate that, in Y. pseudotuberculosis, the transcription of the psaE and psaF genes is constitutive. Alkaline phosphatase fusion studies confirm the topology prediction that PsaE and PsaF are both inner-membrane-associated proteins. PsaE consists of an N-terminal cytoplasmic domain, containing sequence similarity to transcriptional regulators found in two-component systems as well as to the Salmonella typhimurium HilA protein, with a C-terminal domain that is periplasmically localized. PsaF is predicted to be oriented with most of the protein in the periplasm, the hydrophobic N-terminus being either integrated in the inner membrane or cleaved as a signal peptide.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1365-2958.1997.3511719.x
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  • 3
    Electronic Resource
    Electronic Resource
    Ffh and FtsY in a Mycoplasma mycoides signal-recognition particle pathway: SRP RNA and M domain of Ffh are not required for stimulation of GTPase activity in vitro (1997)
    Oxford BSL : Blackwell Publishing Ltd
    Molecular microbiology 24 (1997), S. 0 
    Details
    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Mycoplasma mycoides contains a signal-recognition particle (SRP) composed of an RNA molecule and an SRP54 homologue (Ffh). We have now identified a mycoplasma homologue to the α subunit of the mammalian SRP receptor and Escherichia coli FtsY. The protein (MmFtsY) was expressed in E. coli and purified to homogeneity. MmFtsY has a weak intrinsic GTPase activity but GTP hydrolysis was markedly stimulated when it was combined with mycoplasma Ffh (MmFfh) and SRP RNA. Also, in the absence of SRP RNA GTPase activity was significantly enhanced. Furthermore, GTP hydrolysis was stimulated when MmFtsY was combined with the N-terminal GTPase domain (N+G) of MmFfh. These findings indicate that basic features of the GTPase activation mechanism are independent of the C-terminal M domain of the MmFfh protein. We propose that the activation is mediated to a large extent by contacts between the GTPase domains of the mycoplasma Ffh and FtsY proteins and that the contribution of the M domain and SRP RNA in the activation mechanism is mainly for modifying the conformation of the MmFfh GTPase domain.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1365-2958.1997.3551729.x
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  • 4
    Electronic Resource
    Electronic Resource
    The cioAB genes from Pseudomonas aeruginosa code for a novel cyanide-insensitive terminal oxidase related to the cytochrome bd quinol oxidases (1997)
    Oxford BSL : Blackwell Publishing Ltd
    Molecular microbiology 24 (1997), S. 0 
    Details
    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: The structural genes for the cyanide-insensitive terminal oxidase (CIO) of Pseudomonas aeruginosa were sequenced. The locus comprised two open reading frames, cioA and cioB, coding for gene products of 488 and 335 amino acid residues with predicted molecular masses of 54 241 and 37 016 Da respectively. These genes were encoded by a 2.7 kb transcript and probably comprise an operon. Upstream of a major transcriptional start site is a −10 promoter region and, approximately at nucleotides −50 and +13, there are sequences homologous to the binding site of the transcriptional regulator Anr. The deduced amino acid sequences of CioA and CioB are homologous to the cytochrome bd quinol oxidases of Escherichia coli and Azotobacter vinelandii. However, no cytochrome d-like signals were found in wild-type P. aeruginosa strains. An atypical cytochrome d-like signal was seen under low-aeration growth conditions but only in strains in which the cioAB genes were present on a high-copy-number plasmid. The appearance of these cytochrome d-like signals was not paralleled by a concomitant increase in CIO activity. These data support the hypothesis that the CIO of P. aeruginosa does not contain haem d. This raises the possibility that there is a family of bacterial quinol oxidases related to the cytochrome bd of E. coli that can differ in their haem composition from the E. coli paradigm.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1365-2958.1997.3561728.x
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  • 5
    Electronic Resource
    Electronic Resource
    Sequence requirements for the termination of rolling-circle replication of plasmid pT181 (1997)
    Oxford BSL : Blackwell Publishing Ltd
    Molecular microbiology 24 (1997), S. 0 
    Details
    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Most small multicopy plasmids of Gram-positive bacteria and many in Gram-negative bacteria replicate by a rolling-circle (RC) mechanism. The replication initiator proteins encoded by the RC plasmids and single-stranded bacteriophages of Escherichia coli have origin-specific nicking-closing activities that are required for the initiation and termination of RC replication. We have investigated the sequence requirements for termination of RC replication of plasmid pT181. The initiator nick site is located in the loop of a hairpin region (IRII) within the pT181 origin of replication. By mutational analysis, we have found that several nucleotides within the stem of IRII which are critical for the initiation activity are dispensable for termination of replication. We also demonstrate that nucleotides in the right arm of IRII, but not the left arm, are absolutely required for termination of RC replication. We have also identified specific nucleotides in IRII that are critical for its termination activity. The sequence of the right arm of the hairpin must be located downstream of the initiator nick site for termination, suggesting that termination requires a specific orientation of the initiator protein at the origin.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1365-2958.1997.3641730.x
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  • 6
    Electronic Resource
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    Modulator protein RsbR regulates environmental signalling in the general stress pathway of Bacillus subtilis (1997)
    Oxford BSL : Blackwell Publishing Ltd
    Molecular microbiology 24 (1997), S. 0 
    Details
    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Bacillus subtilis responds to signals of environmental and metabolic stress by inducing over 40 general stress genes under the control of the σB transcription factor. σB activity is regulated post-translationally by a multicomponent network composed of two coupled partner-switching modules, RsbX-RsbS-RsbT and RsbU-RsbV-RsbW, each containing a serine phosphatase (X or U), an antagonist protein (S or V), and a switch protein/serine kinase (T or W). The upstream module (X-S-T) is required to transmit signals of environmental stress. In contrast, the downstream module (U-V-W) is required to transmit signals of energy stress as well as the environmental signals conveyed to it from the upstream module. Until now the function of the rsbR gene product was unknown. RsbR shares significant sequence similarity with the RsbS and RsbV antagonist proteins whose phosphorylation states control key protein–protein interactions within their respective modules. Here we present evidence that RsbR is associated with RsbS in the upstream, environmental-sensing module. To investigate RsbR function, we constructed deletion and point mutations within rsbR and tested their effects on expression of σB-dependent reporter fusions, both singly and in combination with other rsb mutations. To determine the possible interaction of RsbR with other Rsb proteins, we tested the ability of wild-type or mutant RsbR to activate transcription in the yeast two-hybrid system in conjunction with other Rsb regulators. On the basis of this genetic analysis, we conclude that RsbR is a positive regulator which modulates σB activity in response to salt and heat stress. Our data further suggest that: (i) RsbR influences the antagonist function of RsbS by direct protein–protein interaction; and (ii) this interaction with RsbS is likely controlled by the phosphorylation state of RsbR.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1365-2958.1997.3631732.x
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  • 7
    Electronic Resource
    Electronic Resource
    Analysis of the architecture of the transcription factor σN (σ54) and its domains by circular dichroism (1997)
    Oxford BSL : Blackwell Publishing Ltd
    Molecular microbiology 24 (1997), S. 0 
    Details
    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Enhancer-dependent transcription in bacteria requires the alternative transcription factor σN (σ54), which forms an RNA polymerase holoenzyme that binds promoters as a transcriptionally inactive complex. We have examined the structure of σN by circular dichroism (CD) analysis. The σN protein and its domains are well structured in the absence of the core RNA polymerase subunits or promoter DNA. Denaturation of σN by temperature as followed by changes in CD shows a concomitant loss of secondary and tertiary structures with a melting temperature of 36°C. The secondary structure displays a two-state melting curve with a second Tm of 85°C. The amino-terminal Region I activation domain together with the acidic Region II does not contribute to the two-state melting. In marked contrast, the integrity of the C-terminal DNA-binding domain is required for the two-state melting. Measurements of pKb also demonstrated that a C-terminal part of σN, but not regions I or I + II, is required for the structural integrity of σN at high pH. Measurements of pKa suggested that α-helical structures are important in σN for the establishment of tertiary structural elements. The tertiary structure near ultraviolet CD signals of σN do not require regions I or I + II but were strongly diminished by C-terminal truncation of σN. Promoter DNA binding resulted in aconformational change in σN, permitting the determination of a binding constant. A typical B-DNA conformation was adopted by the promoter DNA. Implications for the modular domain organization of σN, the function of C-terminal sequences, and domain communication and its role in activation of transcription are discussed.
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    URL: http://dx.doi.org/10.1046/j.1365-2958.1997.3691738.x
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  • 8
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    Transcriptional regulation of the macrophage-induced gene (gspA) of Legionella pneumophila and phenotypic characterization of a null mutant (1997)
    Oxford BSL : Blackwell Publishing Ltd
    Molecular microbiology 24 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Expression of the global stress protein gene (gspA) is induced during the intracellular infection of macrophages and upon exposure of Legionella pneumophila to in vitro stress stimuli. Transcription of gspA is regulated by two promoters, one of which is regulated by the σ32 heat-shock transcription factor. We utilized a gspA promoter fusion to a promoterless lacZ to probe the phagososmal ‘microenvironment’ for the kinetics of exposure of intracellular L. pneumophila to stress stimuli. Expression through the gspA promoter was constitutively induced by approx. 16-fold throughout the intracellular infection, and occurred predominantly through the σ32-regulated promoter. Expression of the gspA promoter was induced approx. 4.5-fold, 5-, 11- and 9-fold upon exposure of L. pneumophila to heat shock, oxidative stress, acid shock, and osmotic shock, respectively. An isogenic insertion mutant of L. pneumophila in gspA (strain AA224) was constructed by allelic exchange in the wild-type strain AA200. Compared to in vitro-grown wild-type strain AA200, AA224 was more susceptible to all four in vitro stress stimuli. The wild-type phenotypes were restored to strain AA224 by complementation with a plasmid containing wild-type gspA. There was no difference between the wild-type strain and the gspA mutant in cytopathogenicity to U937 cells or in their kinetics of intracellular replication within macrophages and amoebae. However, compared to in vitro-grown bacteria, macrophage-grown and amoebae-grown AA200 and AA224 showed an equal and dramatic increase in resistance to in vitro stress stimuli. Our data showed that regardless of the capacity of L. pneumophila to subvert the microbicidal mechanisms of the macrophage, intracellular L. pneumophila is exposed to a high level of stress stimuli throughout the intracellular infection. Although the GspA protein is required for protection of the bacteria against in vitro stress stimuli, and is induced during intracellular multiplication, the loss of its function is probably compensated for by other macrophage-induced and stress-induced proteins within the intracellular environment.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1365-2958.1997.3661739.x
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  • 9
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    Studies on prolysostaphin processing and characterization of the lysostaphin immunity factor (Lif) of Staphylococcus simulans biovar staphylolyticus (1997)
    Oxford, UK : Blackwell Publishing Ltd
    Molecular microbiology 23 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Lysostaphin is an extracellular glycylglycine endopep-tidase produced by Staphylococcus simulans biovar staphylolyticus ATCC1362 that lyses staphylococcal cells by hydrolysing the polyglycine interpeptide bridges of the peptidoglycan. Renewed analysis of the sequence of the lysostaphin gene (Iss), and the sequencing of the amino-terminus of purified prolysostaphin and of mature lysostaphin revealed that lysostaphin is organized as a preproprotein of 493 amino acids (aa), with a signal peptide consisting of 36 aa, a propeptide of 211 aa from which 195 aa are organized in 15 tandem repeats of 13 aa length, and a mature protein of 246 aa. Prolysostaphin is processed in the culture supernatant of S. simulans biovar staphylolyticus by an extracellular cysteine protease. Although prolysostaphin was staphylolytically active, the mature lysostaphin was about 4.5-fold more active. The controlled expression in Staphylococcus carnosus of Iss and Iss with deletions in the prepropeptide region indicated that the tandem repeats of the propeptide are not necessary for protein export or activation of Lss, but keep Lss in a less active state. Intracellular expressed pro- and mature lysostaphin exert staphy-lolytic activity in cell-free extracts, but do not affect growth of the corresponding clones. We characterized a lysostaphin immunity factor gene (lif) which is located in the opposite direction to Iss. The expression of lif in S. carnosus led to an increase in the serine/glycine ratio of the interpeptide bridges of peptidoglycan from 2 to 35%, suggesting that lysostaphin immunity depends on serine incorporation into the interpeptide bridge. If, in addition to lif, Iss is co-expressed the serine/glycine ratio is further increased to 58%, suggesting that Lss selects for optimal serine incorporation. Lif shows similarity to FemA and FemB
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1365-2958.1997.2911657.x
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  • 10
    Electronic Resource
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    Catabolite repression of the Bacillus subtilis gnt operon exerted by two catabolite-responsive elements (1997)
    Oxford, UK : Blackwell Publishing Ltd
    Molecular microbiology 23 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Catabolite repression of Bacillus subtilis catabolic operons is supposed to occur via a negative regulatory mechanism involving the recognition of a cis-acting catabolite-responsive element (cre) by a complex of CcpA, which is a member of the GalR-LacI family of bacterial regulatory proteins, and the seryl-phos-phorylated form of HPr (P-ser-HPr), as verified by recent studies on catabolite repression of the gnt operon. Analysis of the gnt promoter region by deletions and point mutations revealed that in addition to the ere in the first gene (gntR) of the gnt operon (credown), this operon contains another ere located in the promoter region (creup). A translational gntR-lacZ fusion expressed under the control of various combinations of wild-type and mutant credown and creup was integrated into the chromosomal amyE locus, and then catabolite repression of p-galac-tosidase synthesis in the resultant integrants was examined. The in vivo results implied that catabolite repression exerted by creup was probably independent of catabolite repression exerted by credown; both creup and credown catabolite repression involved CcpA. Catabolite repression exerted by creup was independent of P-ser-HPr, and catabolite repression exerted by credown was partially independent of P-ser-HPr. DNase I footprinting experiments indicated that a complex of CcpA and P-ser-HPr did not recognize creup, in contrast to its specific recognition of credown. However, CcpA complexed with glucose-6-phosphate specifically recognized creup as well as credown, but the physiological significance of this complexing is unknown.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1365-2958.1997.2921662.x
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  • 11
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    A two-gene ABC-type transport system that extrudes Na+ in Bacillus subtilis is induced by ethanol or protonophore (1997)
    Oxford, UK : Blackwell Publishing Ltd
    Molecular microbiology 23 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: A transposition mutant of Bacillus subtilis (designated JC901) that was isolated on the basis of growth inhibition by Na at elevated pH, was deficient in energy-dependent Na extrusion. The capacity of the mutant JC901 for Na -dependent pH homeostasis was unaffected relative to the wild-type strain, as assessed by regulation of cytoplasmic pH after an alkaline shift. The site of transposition was near the 3 -terminal end of a gene, natB, predicted to encode a membrane protein, NatB. NatB possesses six putative membrane-spanning regions at its C-terminus, and exhibits modest sequence similarity to regions of eukaryotic Na+/H+ exchangers. Sequence and Northern blot analyses suggested that natB forms an operon with an upstream gene, natA. The predicted product of natA is a member of the family of ATP-binding proteins that are components of transport systems of the ATP-binding cassette (ABC) or traffic ATPase type. Expression of the lacZ gene that was under control of the promoter for natAB indicated that expression of the operon was induced by ethanol and the protonophore carbonylcyanide p-chlorophenylhydrazone (CCCP), and, more modestly, by Na+, and K+, but not by choline or a high concentration of sucrose. Restoration of the natAB genes, cloned in a recombinant plasmid (pJY1), complemented the Na+-sensitive phe-notype of the mutant JC901 at elevated pH and significantly increased the resistance of the mutant to growth inhibition by ethanol and CCCP at pH 7; ethanol was not excluded, however, from the cells expressing natAB, so ethanol-resistance does not result from NatAB-dependent ethanol efflux. Transformation of the mutant with pJY1 did markedly enhance the capacity for Na+
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1365-2958.1997.2951656.x
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  • 12
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    DnaA protein stimulates polA gene expression in Escherichia coli (1997)
    Oxford, UK : Blackwell Publishing Ltd
    Molecular microbiology 23 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: The polA gene of Escherichia coli encodes DNA polymerase I that is involved in DNA replication and repair. Despite the wide knowledge about structure and function of DNA polymerase I, there is little insight into the regulatory mechanisms involved in polA expression. DnaA is the initiator protein for DNA replication in E. coli. There are two putative DnaA-binding sites within the extended promoter region of polA. In this work we studied the influence of altered levels of DnaA protein on polA expression. We found that DnaA overproduction increases polA expression in stationary-phase cultures. The stimulation effect was independent of rpoS, which encodes the sigma factor for stationary-phase-inducible genes. However, it was modulated by ppGpp. Comparative S1 analyses revealed that the induction was based on transcriptional stimulation. Footprint-ing experiments demonstrated that DnaA binds only to the proximal DnaA box near the polA promoter. These results suggest an additional role for DnaA as transcriptional activator of polA at least under certain physiological conditions.
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    URL: http://dx.doi.org/10.1046/j.1365-2958.1997.2961658.x
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  • 13
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    Molecular characterization of the Bacillus anthracis main S-layer component: evidence that it is the major cell-associated antigen (1997)
    Oxford, UK : Blackwell Publishing Ltd
    Molecular microbiology 23 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Bacillus anthracis, the aetiological agent of anthrax, is a Gram-positive spore-forming bacterium. The cell wall of vegetative cells of B. anthracis is surrounded by an S-layer. An array remained when sap, a gene described as encoding an S-layer component, was deleted. The remaining S-layer component, termed EA1, is chromosomally encoded. The gene encoding EA1 (eag) was obtained on two overlapping fragments in Escherichia coli and shown to be contiguous to the sap gene. The EA1 amino acid sequence, deduced from the eag nucleotide sequence, shows classical S-layer protein features (no cysteine, only 0.1% methionine, 10% lysine, and a weakly acidic pi). Similar to Sap and other Gram-positive surface proteins, EA1 has three 'S-layer-homology’motifs immediately downstream from a signal peptide. Single- and double-disrupted mutants were constructed. EA1 and Sap were co-localized at the cell surface of the wild-type bacilli. However, EA1 was more tightly bound than Sap to the bacteria. Electron microscopy studies and in vivo experiments with the constructed mutants showed that EA1 constitutes the main lattice of the B. anthracis S-layer, and is the major cell-associated antigen.
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    URL: http://dx.doi.org/10.1046/j.1365-2958.1997.2941659.x
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  • 14
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    Random shuttle mutagenesis: gonococcal mutants deficient in pilin antigenic variation (1997)
    Oxford, UK : Blackwell Publishing Ltd
    Molecular microbiology 23 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Shuttle mutagenesis has been adapted to randomly mutate the genome of Neisseria gonorrhoeae (gono-coccus; Gc). A size-restricted plasmid library of Gc strain FA1090 was mutated with the mini-transposon mTnEGNS. Randomness was tested by checking for transposon insertion bias between vector and insert DNA, Gc transformation efficiency of individual mutated clones, and representation of unique clones before and after Gc transformation with a mutated pool of DNA. Mutants created by random shuttle mutagenesis were screened, using a colony-based polymerase chain reaction assay, for the ability to undergo pilin antigenic variation. Out of 8064 mutants screened, 22 unique transposon insertion mutants were found to be antigenic variation deficient (Avd). The Avd mutants were separated into five types according to recombination defect-associated phenotypes, including colony growth, natural DNA transformation competence, and repair of DNA damage caused by ultraviolet radiation.
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    URL: http://dx.doi.org/10.1046/j.1365-2958.1997.2971660.x
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  • 15
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    Replication of minichromosomes in a host in which chromosome replication is random (1997)
    Oxford, UK : Blackwell Publishing Ltd
    Molecular microbiology 23 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Minichromosomes are plasmids with the origin of chromosome replication, oriC, as their only origin of replication. In Escherichia coli, minichromosomes are compatible with the chromosome and replicate in a cell-cycle-specific manner at the same time as oriC located on the chromosome initiates replication. In int strains, oriC has been inactivated and replaced by a plasmid origin. Because plasmids control their own replication, chromosome replication is uncoupled from the normal cell-cycle control and is random with respect to the cell cycle in the int strains. We have used an intP1 strain to address the question of whether minicromosome replication is coupled to the replication of the chromosome or is governed by cell-cycle-specific signals. Minichromosome replication was analysed by density-shift experiments and found not to be random in the randomly replicating intP1 host. This suggests that the cell-cycle-specific control functions of oriC replication are operating also in the intP1 strain.
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    URL: http://dx.doi.org/10.1046/j.1365-2958.1997.2981663.x
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  • 16
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    Maltose-binding protein interacts simultaneously and asymmetrically with both subunits of the Tar chemoreceptor (1997)
    Oxford, UK : Blackwell Publishing Ltd
    Molecular microbiology 23 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: The Tar chemotactic signal transducer of Escherichia coli mediates attractant responses to L-aspartate and to maltose. Aspartate binds across the subunit interface of the periplasmic receptor domain of a Tar homodimer. Maltose, in contrast, first binds to the periplasmic maltose-binding protein (MBP), which in its ligand-stabilized closed form then interacts with Tar. Intragenic complementation was used to determine the MBP-binding site on the Tar dimer. Mutations causing certain substitutions at residues Tyr-143, Asn-145, Gly-147, Tyr-149, and Phe-150 of Tar lead to severe defects in maltose chemotaxis, as do certain mutations affecting residues Arg-73, Met-76, Asp-77, and Ser-83. These two sets of mutations defined two complementation groups when the defective proteins were co-expressed at equal levels from compatible plasmids. We conclude that MBP contacts both subunits of the Tar dimer simultaneously and asymmetrically. Mutations affecting Met-75 could not be complemented, suggesting that this residue is important for association of MBP with each subunit of the Tar dimer. When the residues involved in interaction with MBP were mapped onto the crystal structure of the Tar periplasmic domain, they localized to a groove at the membrane-distal apex of the domain and also extended onto one shoulder of the apical region.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1365-2958.1997.3001661.x
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  • 17
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    Gzf3p, a fourth GATA factor involved in nitrogen-regulated transcription in Saccharomyces cerevisiae (1997)
    Oxford, UK : Blackwell Publishing Ltd
    Molecular microbiology 23 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: In Saccharomyces cerevisiae, two positive transcription factors of the GATA family, Gln3p and NiMp/ Gatlp, upregulate the expression of multiple nitrogen pathway genes via upstream 5-GATA-3′ sequences. Another GATA factor, Uga43p/Da180p, downregulates to varying degrees the expression of some nitrogen-regulated genes. Here, we report the functional analysis of a fourth GATA factor, Gzf 3p/Ni12p, whose gene was discovered by systematic sequencing of chromosome X. The Gzf3 protein most closely resembles Uga43p. Similar to Uga43p, Gzf3p has the properties of a negative GATA factor. While Uga43p is active specifically under nitrogen-derepression conditions, Gzf 3p exerts its negative regulatory function specifically on preferred nitrogen sources: it is involved in nitrogen repression of NiMp-dependent transcription. At least one positive GATA factor is required for the UGA43 and GZF3 genes to be expressed. The Uga43p factor negatively regulates GZF3 expression and vice versa. In addition, both Uga43p and Gzf3p moderately regulate expression of their own genes. These two proteins seem to be parts of a complex network of GATA factors which probably play a determining role in nitrogen-regulated transcription.
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    Different structures of selected and unselected araB-lacZ fusions (1997)
    Oxford, UK : Blackwell Publishing Ltd
    Molecular microbiology 23 (1997), S. 0 
    Details
    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Formation of araB-lacZ coding-sequence fusions is a key adaptive mutation system. Eighty-four independent araB-lacZ fusions were sequenced. All fusions carried rearranged MuR linker sequences between the araB and lacZ domains indicating that they arose from the standard intermediate of the well-characterized Mu DNA rearrangement process, the strand transfer complex (STC). Five non-standard araB-lacZ fusions isolated after indirect sib selection had novel structures containing back-to-back inverted MuR linkers. The observation that different isolation procedures gave rise to standard and non-standard fusions indicates that cellular physiology can influence late steps in the multi-step biochemical sequence leading to araB-lacZ fusions. Each araB-lacZ fusion contained two novel DNA junctions. The MuR-lacZ junctions showed‘hot-spotting’according to established rules for Mu target selection. The araB-MuR and MuR-MuR junctions all involved exchanges at regions of short sequence homology. More extensive homology between MuR and araB sequences indicates potential STC isomerization into a resolvable four-way structure analogous to a Holliday junction. These results highlight the molecular complexity of araB-lacZ fusion formation, which may be thought of as a multi-step cell biological process rather than a unitary biochemical reaction.
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    A novel regulatory system required for pathogenicity of Xanthomonas campestris is mediated by a small diffusible signal molecule (1997)
    Oxford BSL : Blackwell Publishing Ltd
    Molecular microbiology 24 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Mutations in the seven clustered rpf genes cause downregulated synthesis of extracellular enzymes and reduced virulence of Xanthomonas campestris pathovar campestris (Xcc). The phenotype of mutants in one of the genes, rpfF, can be restored by a diffusible extracellular factor (DSF) produced by all Xcc strains tested, apart from rpfF and rpfB mutants. DSF accumulates in early stationary phase (when synthesis of enzymes is maximal), but levels decline subsequently. Addition of DSF to exponentially-growing wild-type bacteria does not cause precocious enzyme synthesis. rpfB and rpfF are expressed throughout growth, but the rate increases in early stationary phase. RpfB is predicted to be a long-chain fatty acyl CoA ligase, and RpfF shows some relatedness to enoyl CoA hydratases. The properties of DSF suggest that it may be a fatty-acid derivative, and certain lipid preparations possess DSF activity at higher concentrations. These include lipid extracts and acid-hydrolysed lipopolysaccharide and lipid A from Xcc, and purified dodecanoic and hydroxydodecanoic acid. DSF production is confined to certain xanthomonads. We propose a model for the DSF system, which represents a novel mechanism for regulating virulence factor synthesis in response to physiological or environmental changes.
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    A C-terminal di-leucine motif and nearby sequences are required for NH4+-induced inactivation and degradation of the general amino acid permease, Gap1p, of Saccharomyces cerevisiae (1997)
    Oxford BSL : Blackwell Publishing Ltd
    Molecular microbiology 24 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: The general amino acid permease, Gap1, of Saccharomyces cerevisiae is very active in cells grown on proline as the sole nitrogen source. Adding NH4+ to the medium triggers inactivation and degradation of the permease via a regulatory process involving Npi1p/Rsp5p, a ubiquitin–protein ligase. In this study, we describe several mutations affecting the C-terminal region of Gap1p that render the permease resistant to NH4+-induced inactivation. An in vivo isolated mutation (gap1pgr ) causes a single Glu→Lys substitution in an amino acid context similar to the DXKSS sequence involved in ubiquitination and endocytosis of the yeast α-factor receptor, Ste2p. Another replacement, substitution of two alanines for a di-leucine motif, likewise protects the Gap1 permease against NH4+-induced inactivation. In mammalian cells, such a motif is involved in the internalization of several cell-surface proteins. These data provide the first indication that a di-leucine motif influences the function of a plasma membrane protein in yeast. Mutagenesis of a putative phosphorylation site upstream from the di-leucine motif altered neither the activity nor the regulation of the permease. In contrast, deletion of the last eleven amino acids of Gap1p, a region conserved in other amino acid permeases, conferred resistance to NH4+ inactivation. Although the C-terminal region of Gap1p plays an important role in nitrogen control of activity, it was not sufficient to confer this regulation to two NH4+-insensitive permeases, namely the arginine (Can1p) and uracil (Fur4p) permeases.
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    HlyX, the FNR homologue of Actinobacillus pleuropneumoniae, is a [4Fe–4S]-containing oxygen-responsive transcription regulator that anaerobically activates FNR-dependent Class I promoters via an enhanced AR1 contact (1997)
    Oxford BSL : Blackwell Publishing Ltd
    Molecular microbiology 24 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: The hlyX gene of the pig pathogen Actinobacillus pleuropneumoniae encodes HlyX, a homologue of FNR, the anaerobic transcription regulator of Escherichia coli. The hlyX gene complements the anaerobic respiratory deficiencies of E. coli fnr mutants but also induces the expression of an otherwise latent haemolysin. Therefore, FNR and HlyX have distinct but overlapping regulons. The hlyX gene has been overexpressed as a gst ::hlyX fusion and the HlyX protein purified. Similar to FNR, HlyX can acquire a [4Fe–4S] cluster, which promotes binding to the FNR box (Kd of 20–30 nM) under anaerobic conditions. Expression of hlyX in E. coli induced the anaerobic production of at least five polypeptides, including the yfiD gene product, which were not induced by fnr. Analysis of the yfiD promoter region revealed the presence of two FNR boxes situated at −61.5 and −114.5. Consistent with this observation, expression from the semi-synthetic Class I promoter FF+20pmelR was efficiently activated by HlyX but not by FNR. The weaker level of FNR-mediated activation of Class I promoters suggests that there is a poorer activating contact (activating region 1 (AR1) equivalent) between FNR and RNA polymerase at these promoters and that HlyX possesses an additional or improved AR1. The AR1 of HlyX is partially characterized by a surface-exposed region around amino acid A187, which confers the altered specificity and provides an explanation for the existence of distinct but overlapping HlyX and FNR regulons.
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    URL: http://dx.doi.org/10.1046/j.1365-2958.1997.3801737.x
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    Genetic evidence of separate repressor and activator activities of the XylR regulator of the TOL plasmid, pWWO, of Pseudomonas putida (1997)
    Oxford, UK : Blackwell Publishing Ltd
    Molecular microbiology 23 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: The XylR protein encoded by pWWO, the TOL (toluene biodegradation) plasmid of Pseudomonas putida, activates at a distance the transcription of Pu and Ps, which are the two σ54-dependent promoters of the plasmid, but it also downregulates its own σ70-promoter, Pr, which divergently overlaps the upstream activating sites of Ps. All regulatory elements that control Pr activity have been faithfully reproduced in Escherichia coli, and the basis of the autoregulation of XylR transcription has been examined by monitoring the activity in vivo of different combinations of mutant proteins and promoters in rpoN+ and rpoN- genetic backgrounds. By using PsIPr regions bearing deleted or offset binding sites for XylR and the σ54-containing RNA polymerase, we could show that formation of a nucleoprotein complex involving the polymerase bound to the divergent promoter Ps is not required for downregulation of Pr. Mutant XylR proteins, G268N and A311V (mutated within the NTP-binding region of XylR) or R453H (affected in multi-merization), which are unable to activate (-dependent transcription from Ps, were indistinguishable from the wild-type XylR in their ability to repress a reporter Pr-lacZ fusion. Autoregulation of XylR is therefore due exclusively to the binding of the protein to its target sites at the Pr promoter. This allows one to define sensu stricto XylR as a transcriptional repressor, independently of its activator role in other promoters.
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    URL: http://dx.doi.org/10.1046/j.1365-2958.1997.3091673.x
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    Pathogenicity islands of virulent bacteria: structure, function and impact on microbial evolution (1997)
    Oxford, UK : Blackwell Publishing Ltd
    Molecular microbiology 23 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Virulence genes of pathogenic bacteria, which code for toxins, adhesins, invasins or other virulence factors, may be located on transmissible genetic elements such as transposons, plasmids or bacteriophages. In addition, such genes may be part of particular regions on the bacterial chromosome, termed‘pathogenicity islands’(Pais). Pathogenicity islands are found in Gram-negative as well as in Gram-positive bacteria. They are present in the genome of pathogenic strains of a given species but absent or only rarely present in those of non-pathogenic variants of the same or related species. They comprise large DNA regions (up to 200 kb of DNA) and often carry more than one virulence gene, the G+C contents of which often differ from those of the remaining bacterial genome. In most cases, Pais are flanked by specific DNA sequences, such as direct repeats or insertion sequence (IS) elements. In addition, Pais of certain bacteria (e.g. uropathogenic Escherichia coli, Yersinia spp., Helicobacter pylori) have the tendency to delete with high frequencies or may undergo duplications and amplifications. Pais are often associated with tRNA loci, which may represent target sites for the chromosomal integration of these elements. Bacteriophage attachment sites and cryptic genes on Pais, which are homologous to phage integrase genes, plasmid origins of replication or IS elements, indicate that these particular genetic elements were previously able to spread among bacterial populations by horizontal gene transfer, a process known to contribute to microbial evolution.
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    Autoregulation of the Escherichia coli replication initiator protein, DnaA, is indirect (1997)
    Oxford, UK : Blackwell Publishing Ltd
    Molecular microbiology 23 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: The expression of dnaA is autoregulated, in that transcription of the gene increases when DnaA is inactivated (and initiation of replication prevented) and decreases when DnaA is supplied in excess. However, the inactivation of DnaA does not necessarily lead to increased DnaA production, as dnaA(7s; temperature sensitive) strains which are integratively suppressed by derivatives of the plasmid R1 do not show temperature-induced derepression. Several possible explanations for this unanticipated behaviour were considered and ruled out. We suggest here that the completion of a critical step in initiation may prevent dnaA derepression: although DnaA would be required to complete this step at oriC, DnaA(Ts) would be sufficient at the R1 origin. Autoregulation of dnaA has been attributed to the binding of DnaA at a consensus binding site in the dnaA promoter region. We show here, using reporter systems, that this DnaA-binding site is not required for the autoregu-latory response. We find, further, that replacement of the chromosomal dnaA gene with one containing a mutated binding site causes no demonstrable pheno-typic change: cells with the mutant gene show no disadvantage in competition with dnaA+ cells.
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    Double-glycine-type leader peptides direct secretion of bacteriocins by ABC transporters: colicin V secretion in Lactococcus lactis (1997)
    Oxford, UK : Blackwell Publishing Ltd
    Molecular microbiology 23 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Many non-lantibiotic bacteriocins of lactic acid bacteria are produced as precursors which have N-terminal leader peptides that share similarities in amino acid sequence and contain a conserved processing site of two glycine residues in positions -1 and -2. A dedicated ATP-binding cassette (ABC) transporter is responsible for the proteolytic cleavage of the leader peptides and subsequent translocation of the bacteriocins across the cytoplasmic membrane. To investigate the role that these leader peptides play in the recognition of the precursor by the ABC transporters, the leader peptides of leucocin A, lactococcin A or colicin V were fused to divergicin A, a bacteriocin from Carnobacterlum divergens that is secreted via the cell's general secretion pathway. Production of divergicin was monitored when these fusion constructs were introduced into Leuconostoc gelidum, Lactococcus lactis and Escherichia coli, which carry the secretion apparatus for leucocin A, lactococcins A and B, and colicin V, respectively. The different leader peptides directed the production of divergicin in the homologous hosts. In some cases production of divergicin was also observed when the leader peptides were used in heterologous hosts. For ABC-transporter-dependent secretion in E. coli the outer membrane protein TolC was required. Using this strategy, colicin V was produced in L. lactis by fusing this bacteriocin behind the leader peptide of leucocin A.
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    RNase E: still a wonderfully mysterious enzyme (1997)
    Oxford, UK : Blackwell Publishing Ltd
    Molecular microbiology 23 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Ribonuclease E (RNase E), which is encoded by an essential Escherichia coli gene known variously as rne, ams, and hmp, was discovered initially as an rRNA-processing enzyme but is now known to have a general role in RNA decay. Multiple functions, including the ability to cleave RNA endonucleolyticaliy in AU-rich single-strand regions, RNA-binding capabilities, and the ability to interact with polynucleotide phosphorylase and other proteins implicated in the processing and degradation of RNA, are encoded by its 1061 amino acid residues. The presence of homologues and functional analogues of the rne gene in a variety of prokaryotic and eukaryotic species suggests that its functions have been highly conserved during evolution. While much has been learned in recent years about the structure and functions of RNase E, there is continuing mystery about possible additional activities and molecular interactions of this enzyme.
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    URL: http://dx.doi.org/10.1111/j.1365-2958.1997.tb02593.x
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    The nucleoside diphosphate kinase of Mycobacterium smegmatis: identification of proteins that modulate specificity of nucleoside triphosphate synthesis by the enzyme (1997)
    Oxford BSL : Blackwell Publishing Ltd
    Molecular microbiology 24 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: We report the purification and characterization of the enzyme nucleoside diphosphate kinase (Ndk) from Mycobacterium smegmatis. The N-terminus of the enzyme was blocked but an internal sequence showed approx. 70% homology with the same enzymes from Pseudomonas aeruginosa and Escherichia coli. Immobilization of the mycobacterial nucleoside diphosphate kinase on a Sepharose 4B matrix and passing the total cell extract through it revealed four proteins (P70, P65, P60, and P50, respectively) of Mr 70 kDa, 65 kDa, 60 kDa and 50 kDa that were retained by the column. While the proteins of Mr 70 kDa and 50 kDa modulated the activity of Ndk directing it towards GTP synthesis, the 60 kDa protein channelled the specificity of Ndk entirely towards CTP synthesis. The 65 kDa protein modulated the specificity of Ndk directing it entirely towards UTP synthesis. The specificity for such mycobacterial proteins towards NTP synthesis is retained when they are complexed with P. aeruginosa Ndk. We further demonstrate that the P70 protein is pyruvate kinase and that each of the four proteins forms a complex with Ndk and alters its substrate specificity. Given the ubiquitous nature of Ndk in the living cell and its role in maintaining correct ratios of intracellular nucleoside triphosphates, the implications of the occurrence of these complexes have been discussed in relation to the precursor pool for cell wall biosynthesis as well as RNA/DNA synthesis.
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    The C-terminal domain of the secretin PulD contains the binding site for its cognate chaperone, PulS, and confers PulS dependence on pIVf1 function (1997)
    Oxford BSL : Blackwell Publishing Ltd
    Molecular microbiology 24 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Related outer membrane proteins, termed secretins, participate in the secretion of macromolecules across the outer membrane of many Gram-negative bacteria. In the pullulanase-secretion system, PulS, an outer membrane-associated lipoprotein, is required both for the integrity and the proper outer membrane localization of the PulD secretin. Here we show that the PulS-binding site is located within the C-terminal 65 residues of PulD. Addition of this domain to the filamentous phage secretin, pIV, or to the unrelated maltose-binding protein rendered both proteins dependent on PulS for stability. A chimeric protein composed of bacteriophage f1 pIV and the C-terminal domain of PulD required properly localized PulS to support phage assembly. An in vivo complex formed between the pIV-PulD65 chimera and PulS was detected by co-immunoprecipitation and by affinity chromatography.
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    Structure of the DNA-binding domain of the OmpR family of response regulators (1997)
    Oxford BSL : Blackwell Publishing Ltd
    Molecular microbiology 24 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
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    URL: http://dx.doi.org/10.1046/j.1365-2958.1997.3571723.x
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    Structural basis for differential receptor binding of cholera and Escherichia coli heat-labile toxins: influence of heterologous amino acid substitutions in the cholera B-subunit (1997)
    Oxford BSL : Blackwell Publishing Ltd
    Molecular microbiology 24 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: The closely related B-subunits of cholera toxin (CTB) and Escherichia coli heat-labile enterotoxin (LTB) both bind strongly to GM1 ganglioside receptors but LTB can also bind to additional glycolipids and glycoproteins. A number of mutant CT B-subunits were generated by substituting CTB amino acids with those at the corresponding positions in LTB. These were used to investigate the influence of specific residues on receptor-binding specificity. A mutated CTB protein containing the first 25 residues of LTB in combination with LTB residues at positions 94 and 95, bound to the same extent as native LTB to both delipidized rabbit intestinal cell membranes, complex glycosphingolipids (polyglycosylceramides) and neolactotetraosylceramide, but not to non-GM1 intestinal glycosphingolipids. In contrast, when LTB amino acid substitutions in the 1–25 region were combined with those in the 75–83 region, a binding as strong as that of LTB to intestinal glycosphingolipids was observed. In addition, a mutant LTB with a single Gly-33→Asp substitution that completely lacked affinity for both GM1 and non-GM1 glycosphingolipids could still bind to receptors in the intestinal cell membranes and to polyglycosylceramides. We conclude that the extra, non-GM1 receptors for LTB consist of both sialylated and non-sialylated glycoconjugates, and that the binding to either class of receptors is influenced by different amino acid residues within the protein.
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    A quest for symbiosis-specific genes urges itself upon Rhizobium geneticists (1997)
    Oxford BSL : Blackwell Publishing Ltd
    Molecular microbiology 24 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
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    Isolation and characterization of the mycobacterial phagosome: segregation from the endosomal/lysosomal pathway (1997)
    Oxford BSL : Blackwell Publishing Ltd
    Molecular microbiology 24 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Mycobacteria have the ability to persist within host phagocytes, and their success as intracellular pathogens is thought to be related to the ability to modify their intracellular environment. After entry into phagocytes, mycobacteria-containing phagosomes acquire markers for the endosomal pathway, but do not fuse with lysosomes. The molecular machinery that is involved in the entry and survival of mycobacteria in host cells is poorly characterized. Here we describe the use of organelle electrophoresis to study the uptake of Mycobacterium bovis bacille Calmette Guerin (BCG) into murine macrophages. We demonstrate that live, but not dead, mycobacteria occupy a phagosome that can be physically separated from endosomal/lysosomal compartments. Biochemical analysis of purified mycobacterial phagosomes revealed the absence of endosomal/lysosomal markers LAMP-1 and β-hexosaminidase. Combining subcellular fractionation with two-dimensional gel electrophoresis, we found that a set of host proteins was present in phagosomes that were absent from endosomal/lysosomal compartments. The residence of mycobacteria in compartments outside the endosomal/lysosomal system may explain their persistence inside host cells and their sequestration from immune recognition. Furthermore, the approach described here may contribute to an improved understanding of the molecular mechanisms that determine the intracellular fate of mycobacteria during infection.
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    Characterization of the negative elements involved in silencing the bgl operon of Escherichia coli: possible roles for DNA gyrase, H-NS, and CRP–cAMP in regulation (1997)
    Oxford BSL : Blackwell Publishing Ltd
    Molecular microbiology 24 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: The bgl operon of Escherichia coli is rendered cryptic and uninducible in wild-type cells by the presence of DNA structural elements that negatively regulate transcription. We have carried out a detailed analysis of the sequences implicated in negative regulation. Fine-structure deletion analysis of the upstream sequences showed the presence of at least two elements involved in silencing the promoter. Chemical probing of genomic DNA in vivo showed that a region of dyad symmetry, present upstream of the promoter, is hypersensitive to KMnO4. The hypersensitive region detected corresponds to the potential cruciform structure implicated earlier in negative regulation. Enhancement of transcription from the wild-type promoter, observed in the presence of the gyrase inhibitor novobiocin, was absent in a mutant that carried point mutations in the inverted repeat. This observation suggests that the activation seen in a gyrase mutant is mediated by destabilization of the cruciform because of reduced supercoiling. Deletion of sequences downstream of the potential cruciform also resulted in an increase in transcription, indicating the presence of a second regulatory element. Measurement of transcription from the bgl promoter carrying the deletion, in a strain that has a mutation in the hns gene, indicated that this region is likely to be involved in binding to H-NS or a protein regulated by H-NS, which acts as a non-specific repressor. We also provide evidence which suggests that transcriptional activation by mutations at the cAMP receptor protein (CRP)-binding site is mediated partly by antagonization of the negative effect of H-NS by CRP–cAMP as a result of its increased affinity for the mutant site.
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    Neisseria gonorrhoeae reverse transcriptase activity does not mediate pilin gene conversion (1997)
    Oxford BSL : Blackwell Publishing Ltd
    Molecular microbiology 24 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
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    URL: http://dx.doi.org/10.1046/j.1365-2958.1997.3681707.x
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    Action at a distance for glp repressor control of glpTQ transcription in Escherichia coli K-12 (1997)
    Oxford BSL : Blackwell Publishing Ltd
    Molecular microbiology 24 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: The adjacent, divergently transcribed glpACB and glpTQ operons of Escherichia coli encode the anaerobic glycerol 3-phosphate dehydrogenase and glycerol 3-phosphate transporter/phosphodiesterase, respectively. These operons are negatively controlled by glp repressor binding to operators that overlap the glpA promoter elements. Using DNase I footprinting, three additional operators (OT1–3) were identified at positions +307 to +359 within the glpT coding region. To assess a potential regulatory role for these remote operators in vivo, a glpT–lacZ transcriptional fusion containing all of the glpA and glpT operators was constructed. The response of this fusion to the glp repressor was compared to fusion constructs in which OT1 and OT3 were inactivated, either by deletion or by site-directed mutagenesis. It was found that repression of glpT conferred by binding of glp repressor to glpA operators was increased about three- to fourfold upon introduction of the remote glpT operators. In addition, two integration host factor (IHF) binding sites were identified downstream of the glpT transcriptional start site at positions +15 to +51 and +193 to +227. A regulatory role for IHF was demonstrated by showing that repression of glpT mediated by GlpR was decreased about twofold in strains deficient in IHF and that mutations in IHF1 and/or IHF2 decreased repression about two- to threefold. The effect of IHF was apparent only when the remote operators were present. All of the results are consistent with a model of repression involving GlpR binding simultaneously to the glpA and remote glpT operators, with intervening DNA forming a loop.
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    Polypeptide chain release factors (1997)
    Oxford BSL : Blackwell Publishing Ltd
    Molecular microbiology 24 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Newly synthesized polypeptide chains are released from peptidyl-tRNA when the ribosome encounters a stop signal on mRNA. Extra-ribosomal proteins (release factors) play an essential role in this process. Although the termination process was first discovered in the late 1960s, much of the mechanism has remained obscure. However, important steps have recently been made in both prokaryotic and eukaryotic organisms in unlocking the secrets of this vital stage in protein synthesis. In this review we summarize these advances and focus attention on the remaining areas of uncertainty, particularly with respect to the models that have been proposed for the action of the GTP-hydrolysing termination factors in prokaryotes and eukaryotes, i.e. RF3 and eRF3.
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    The σS level in starving Escherichia coli cells increases solely as a result of its increased stability, despite decreased synthesis (1997)
    Oxford BSL : Blackwell Publishing Ltd
    Molecular microbiology 24 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: The σS level in starving (stationary phase) Escherichia coli cells increases four- to sixfold following growth in a defined or a complex medium. Chemostat-grown cells, subjected to increasing carbon starvation, also become progressively richer in σS content. These increases occur despite reduced transcription of the σS-encoding gene, rpoS, and translation of rpoS mRNA, and result solely from a large increase in the stability of the sigma protein. Previous results, based on rpoS ::lacZ transcriptional and translational fusions, and on methionine incorporation in σS, had suggested increased synthesis of σS in starving cells. Alternative explanations for these results consistent with the conclusions of this paper are discussed.
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    Cross-talk to the genes for Bacillus anthracis capsule synthesis by atxA, the gene encoding the frans-activator of anthrax toxin synthesis (1997)
    Oxford, UK : Blackwell Publishing Ltd
    Molecular microbiology 23 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: The two major virulence factors of Bacillus anthracis are the tripartite toxin and the polyglutamate capsule, which are encoded by genes on the large plasmids, pX01 and pX02, respectively. The genes atxA, located on pX01, and acpA, located on pX02, encode positive frans-acting proteins that are involved in bicarbonate-mediated regulation of toxin and capsule production, respectively. A derivative strain cured of pX01 produced less capsular substance than the parent strain harbouring both pX01 and pX02, and electroporation of the strain cured of pX01 with a plasmid containing the cloned atxA gene resulted in an increased level of capsule production. An acpA-null mutant was complemented by not only acpA but also the atxA gene. The cap region, which is essential for encapsulation, contains three genes capB, capC, and cap A, arranged in that order. The atxA gene stimulated capsule synthesis from the cloned cap region. Transcriptional analysis of cap by RNA slot-blot hybridization and primer-extension analysis revealed that atxA activated expression of cap in trans at the transcriptional level. These results indicate that cross-talk occurs, in which the pX01-located gene, atxA, activates transcription of the cap region genes located on pX02. We identified two major apparent transcriptional start sites, designated P1 and P2, located at positions 731 bp and 625 bp, respectively, upstream of the translation-initiation codon of capB. Transcription initiated from P1 and P2 was activated by both atxA and acpA, and activation appeared to be stimulated by bicarbonate. Deletion analysis of the upstream region of the cap
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    DNA topology in hyperthermophilic archaea: reference states and their variation with growth phase, growth temperature, and temperature stresses (1997)
    Oxford, UK : Blackwell Publishing Ltd
    Molecular microbiology 23 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: In order to address the dynamics of DNA topology in hyperthermophilic archaea, we analysed the topological state of several plasmids recently discovered in Thermococcales and Sulfolobales. All of these plasmids were from relaxed to highly positively super-coiled in vitro, i.e. they exhibited a significant linking excess compared to the negatively supercoiled plasmids from mesophilic organisms (both Archaea and Bacteria). In the two archaeai orders, plasmid linking number (Lk) decreased as growth temperature was lowered from its optimal value, i.e. positively super-coiled plasmids were relaxed whereas relaxed plasmids became negatively supercoiled. Growth temperatures above the optimum correlated with higher positive supercoiling in Sulfolobales (Lk increase) but with relaxation of positive supercoils in Thermococcus sp. GE31. The topological variation of plasmid DNA isolated from cells at different growth phases were found to be species specific in both archaeai orders. In contrast, the direction of topological variation under temperature stress was the same, i.e. a heat shock correlated with an increase in plasmid positive supercoiling, whilst a cold shock induced negative supercoiling. The kinetics of these effects were analysed in Sulfolobales. In both temperature upshift (from 80 to 85C) and downshift (from 80 to 65C), a transient sharp variation of Lk occurred first, and then DNA supercoiling progressively reached levels typical of steady-state growth at the final temperature. These results indicate that DNA topology can change with physiological states and environmental modifications in hyperthermophilic archaea.
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    Cell-type specificity of the Anabaena fdxN-element rearrangement requires xisH and xisl (1997)
    Oxford, UK : Blackwell Publishing Ltd
    Molecular microbiology 23 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: The fdxN element, along with two other DNA elements, is excised from the chromosome during heterocyst differentiation in Anabaena sp. strain PCC 7120. Previous work showed that rearrangement of the fdxN element requires the xisF gene, which encodes a site-specific recombinase, and suggested that at least one other heterocyst-specific factor is involved. Here we report that the xisH and xisl genes are necessary for the heterocyst-specific excision of the fdxN element. Deletion of a 3.2 kb region downstream of the xisF gene blocked the fdxN-element rearrangement in hetero-cysts. The 3.2 kb deletion was complemented by the two overlapping genes xisH and xisl. Interestingly, extra copies of xlsHI on a replicating plasmid resulted in the xisF-dependent excision of the fdxN element in vegetative cells. Therefore, xisHI are involved in the control of cell-type specificity of the fdxN rearrangement. The xisHI genes had no effect on the two other DNA rearrangements. The xisHl-induced excision of the fdxN element produced strains lacking the element and demonstrates that the 55 kb element contains no essential genes. xisH and xisl do not show similarity to any known genes.
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    Temperature-regulated mRNA accumulation and stabilization for fatty acid desaturase genes in the cyanobacterium Synechococcus sp. strain PCC 7002 (1997)
    Oxford, UK : Blackwell Publishing Ltd
    Molecular microbiology 23 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Cyanobacteria acclimate to low-temperature conditions by desaturating their membrane lipids. The desB (ω3 desaturase) and desC (A9 desaturase) genes of Synechococcus sp. strain PCC 7002 were cloned and characterized, and the expression of the desA (Δ12 desaturase), desB and desC genes was studied as a function of temperature. The steady-state mRNA abundance for the desA gene was threefold higher in cells grown at 22 C than in cells grown at 38°C. des B transcripts were not detected at 38°C, but were abundant in cells grown at 22°C. Levels of desC mRNA were similar at both growth temperatures. The mRNA levels of each desaturase gene increased within 15min of a temperature shift-down to 22°C, and mRNA levels recovered within 15min after a shift-up to 38°C. The cold-induced accumulation of transcripts from the desA and desB genes was suppressed by the addition of chloramphenicol, but the transient elevation of the desC transcript levels at 22°C was not affected by chloramphenicol. The half-lives of the desA and desB mRNAs were significantly longer in cells grown at 22°C than in cells grown at 38°C, but the desC mRNA had a similar half-life at both temperatures. These studies reveal three patterns of temperature regulation for the desaturase genes, whose expression is tightly controlled by a combination of mRNA synthesis and stabilization. These studies demonstrate that elevation of desaturase mRNA levels is not the rate-limiting event during the low-temperature acclimation of cyanobacteria.
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    The Bacillus subtilis chromatin-associated protein Hbsu is involved in DNA repair and recombination (1997)
    Oxford, UK : Blackwell Publishing Ltd
    Molecular microbiology 23 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: The Bacillus subtilis hbs gene encodes an essential chromatin-associated protein termed Hbsu. Hbsu, the counterpart of the Escherichia coli HU protein, binds DNA in a non-specific way but has a clear preference for bent, kinked or altered DNA sequences. To investigate the role of Hbsu in DNA repair and DNA recombination we have constructed a series of site-directed mutants in the hbs gene and used these mutant genes to substitute the wild-type chromosomal hbs gene. The hbs47 mutation, which codes for a mutant protein in which residue Phe-47 has been replaced by Trp, does not cause any discernible phenotype. Additional substitution of residue Arg-55 by Ala (hbs4755 mutation) rendered cells deficient in DNA repair, homologous recombination and (i protein-mediated site-specific recombination. We have also tested the effect on DNA repair of the hbs4755 mutation in combination with mutations in different functions of homologous DNA recombination (recA, recF, recG, recti and addAB). The hbs4755 mutation did not modify the sensitivity of recH and addAB cells to the DNA-damaging agents methylmethane sulphonate (MMS) or 4-nitroquinoline-1-oxide (4NQO), and it only marginally affected recF and recG cells. The hbs4755 mutation blocked intermolecular recombination in recH cells and markedly reduced it (20- to 50-fold) in recF and recG cells, but had no effect on addAB cells. Taken together, these data indicate that the Hbsu protein is required for DNA repair and for homologous DNA recombination.
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    Former prime minister to stand for top WHO post (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 481-482 
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    ISSN: 1546-170X
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Medicine
    Notes: [Auszug] Gro Harlem Brundtland, Norway's former prime minister, has announced that she is willing to stand for election for the post of director-general of the World Health Organization. The UN agency's executive board, made up of health specialists from member states, will vote on the appointment next ...
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    Senate ban on euthanasia splits Australia's doctors (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 479-479 
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    ISSN: 1546-170X
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Medicine
    Notes: [Auszug] Australia's medical community has been deeply divided by the Senate's decision to overturn the world's first law allowing voluntary euthanasia. The Northern Territory's Rights of the Terminally I11 Act 1995, which came into effect last summer, was thrown out by the Senate before Easter in an ...
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    Genetic privacy bill “may hinder research” (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 481-481 
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    ISSN: 1546-170X
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Medicine
    Notes: [Auszug] A bill before Congress that seeks to protect people's genetic privacy has run into unexpected opposition — from members of the science and biotechnology communities. Although scientists strongly support the section of the bill that aims to prevent genetic discrimination in the workplace ...
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    A homologue of Drosophila tissue polarity gene frizzled is expressed in migrating myofibroblasts in the infarcted rat heart (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 541-544 
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    ISSN: 1546-170X
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Medicine
    Notes: [Auszug] Myocardial infarction results in the formation of granulation tissue in the injured ventricular wall. This tissue contains myofibroblasts in highly organized arrays; their contractile properties may help to prevent the infarct area from dilatation. The mechanisms that control myofibroblast ...
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    Sunlight and skin cancer: Inhibition of p53 mutations in UV-irradiated mouse skin by sunscreens (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 510-514 
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    ISSN: 1546-170X
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Medicine
    Notes: [Auszug] UV-induced mutations in the p53 tumor suppressor gene play an essential role in skin cancer development. We report here that such mutations can be detected in UV-irradiated mouse skin months before the gross appearance of skin tumors. Application of SPF-15 sunscreens to mouse skin before each UV ...
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    Protection of chimpanzees from high-dose heterologous HIV-1 challenge by DNA vaccination (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 526-532 
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    ISSN: 1546-170X
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Medicine
    Notes: [Auszug] Novel approaches for the generation of more effective vaccines for HIV-1 are of significant importance. In this report we analyze the immunogenicity and efficacy of an HIV-1 DNA vaccine encoding env, rev and gag/pol in a chimpanzee model system. The immunized animals developed specific cellular and ...
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    Targeting gene expression to hypoxic tumor cells (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 515-520 
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    ISSN: 1546-170X
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Medicine
    Notes: [Auszug] Solid tumors with areas of low oxygen tension (hypoxia) have a poor prognosis, as celts in this environment often survive radiation and chemotherapy. In this report we describe how this hypoxic environment can be used to activate heterologous gene expression driven by a hypoxia-responsive element ...
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    Estrogen inhibits the vascular injury response in estrogen receptor α-deficient mice (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 545-548 
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    ISSN: 1546-170X
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Medicine
    Notes: [Auszug] The atheroprotective effects of estrogen in women are well recognized1, but the underlying mechanisms responsible are not well understood. Blood vessel cells express the classic estrogen receptor, ERα (ref. 2—6), and are directly affected by estrogen, which inhibits the development of ...
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    HIV CCR5 resistance incomplete (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 372-373 
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    ISSN: 1546-170X
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Medicine
    Notes: [Auszug] Hopes that some people might be genetically protected from infection with HIV have been dampened by the findings of researchers in Australia. The findings confirm the fears of many scientists, who had warned people against abandoning safer sex on the basis of these hopes. Last year there was ...
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    Gel microdrop technology for rapid isolation of rare and high producer cells (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 583-585 
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    ISSN: 1546-170X
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Medicine
    Notes: [Auszug] Increasing use of monoclonal antibodies and recombinant proteins has created a demand for methods that accelerate and simplify generation of stable cell lines. Mammalian cell-line development involves isolation or engineering of protein-secreting cells and screening for specificity and high ...
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    HIV-1 antibody : debris or virion? (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 366-367 
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    ISSN: 1546-170X
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Medicine
    Notes: [Auszug] To the editor — Many hypotheses have been put forward to explain the inability of an antibody response to clear or control HIV-1 infection. These include viral variation, lack of efficacy against cell-to-cell spread and the inhibition of activity at certain privileged anatomical sites. We ...
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    Transmission of L. infantum by blood donors (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 368-368 
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    ISSN: 1546-170X
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Medicine
    Notes: [Auszug] To the editor — The possible transmission of Leishmania via blood transfusion was raised by the World Health Organization in 1990. We screen blood from donors to the Monaco Blood Bank who live in areas endemic for L. infantum, checking for potential asymptomatic Leishmania carriers and ...
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    Progression of metastatic human prostate cancer to androgen independence in immunodeficient SCID mice (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 402-408 
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    ISSN: 1546-170X
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Medicine
    Notes: [Auszug] Prostate cancer mortality results from metastasis to bone and hormone-independent tumor growth. Models to study these progressive changes are lacking. Here we describe the propagation of advanced human prostate cancer by direct transfer of surgical samples from patients into immune-deficient male ...
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    Neurotrophic actions of nonimmunosuppressive analogues of immunosuppressive drugs FK506, rapamycin and cyclosporin A (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 421-428 
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    ISSN: 1546-170X
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Medicine
    Notes: [Auszug] We show that the nonimmunosuppressive analogues of the immunosuppressive drugs FK506, rapamycin and cyclosporin A promote neurite outgrowth both in PC12 cells and sensory neu-ronal cultures of dorsal root ganglia with potencies resembling their immunosuppressive homologues. Neurotrophic potencies ...
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    Viral vector-mediated transduction of a modified platelet factor 4 cDNA inhibits angiogenesis and tumor growth (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 437-442 
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    ISSN: 1546-170X
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Medicine
    Notes: [Auszug] Chronic systemic delivery of therapeutic proteins, such as inhibitors of angiogenesis, present a number of difficult pharmacological challenges. To overcome these problems for one such protein, we constructed retroviral and adenoviral vectors that express a novel, secretable form of the ...
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    Elevation of oxygen release by nitroglycerin without an increase in blood flow in the hepatic microcirculation (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 456-459 
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    Source: Nature Archives 1869 - 2009
    Topics: Biology , Medicine
    Notes: [Auszug] The effect of nitroglycerin on oxygen (O2) release in the microcirculation was investigated by examining single, unbranched hepatic sinusoids of rats using dual-spot microspectroscopy. Nitroglycerin significantly increased O2 release from erythrocytes flowing in the sinusoids. Differences in O2 ...
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    Dying in a Japanese Hospital (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 463-464 
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    Source: Nature Archives 1869 - 2009
    Topics: Biology , Medicine
    Notes: [Auszug] by Fumio Yamazaki The Japan Times, ISBN: 4-7890-0845-2 Reviewed by Yusuke Nakamura, Director, Human Genome Center, Laboratory of Molecular Medicine, 1-37-1 Kami-ikebukuro, Tokyo 108, Japan Most terminal cancer patients in Japan have to fight for their lives without knowing the truth; they are ...
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    Histological evidence of fetal pig neural cell survival after transplantation into a patient with Parkinson's disease (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 350-353 
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    ISSN: 1546-170X
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Medicine
    Notes: [Auszug] The movement disorder in Parkinson's disease results from the selective degeneration of a small group of dopaminergic neurons in the substantia nigra pars compacta region of the brain. A number of exploratory studies using human fetal tissue allografts have suggested that transplantation of ...
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    Engineering herpes simplex virus vectors for gene transfer to neurons (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 357-359 
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    ISSN: 1546-170X
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Medicine
    Notes: [Auszug] The delivery and expression of transgenes will require individualized approaches tailored to the specific disease and target tissue. For the central and peripheral nervous system, where neurons are postmitotic and intricate cell–cell relationships and regional specialization complicate ex ...
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    On the market (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 362-364 
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    ISSN: 1546-170X
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Medicine
    Notes: [Auszug] AUTOMATION Interior view of the Mantis bioprocessing system showing the robotic system in close-up with ‘picking’ head in place. The Mantis standalone, multifunctional robotic bioprocessing system from Linear Drives is designed to provide high speed and accurate and automated gridding ...
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    Some UK medical schools make the grade (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 133-133 
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    Source: Nature Archives 1869 - 2009
    Topics: Biology , Medicine
    Notes: [Auszug] The leaders of Britain's medical schools are engaged in an intense round of navel-gazing following a mixed report card on their research efforts that has emerged from a nationwide review of the quality of research in Britain's universities. Organized by the Higher Education Funding Council for ...
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    TNF antibodies get new lease (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 130-130 
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    Source: Nature Archives 1869 - 2009
    Topics: Biology , Medicine
    Notes: [Auszug] Attempts by biotech companies to market antibody-based therapies for septic shock have been repeatedly beaten back by dismal clinical results and a skeptical US Food and Drug Administration (FDA). “It's sepsis 5, biotech 0,” said one stock analyst, referring to the clinical and ...
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    Testing, testing … testing? (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 131-132 
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    Source: Nature Archives 1869 - 2009
    Topics: Biology , Medicine
    Notes: [Auszug] With the commercialization of four genetic tests for breast cancer, it would be reasonable to expect a degree of clarity about risk, prevention and treatment of the disease. Instead, the tests “have opened a Pandora's box” of medical and ethical questions, according to one genetic ...
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    Suppression of SPARC expression by antisense RNA abrogates the tumorigenicity of human melanoma cells (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 171-176 
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    ISSN: 1546-170X
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Medicine
    Notes: [Auszug] Acquisition of invasive/metastatic potential is a key event in tumor progression. Cell surface glycoproteins and their respective matrix ligands have been implicated in this process. Recent evidence reveals that the secreted glycoprotein SPARC (secreted protein, acidic and rich in cysteine) is ...
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    Accommodation of vascularized xenografts: Expression of “protective genes” by donor endothelial cells in a host Th2 cytokine environment (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 196-204 
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    Source: Nature Archives 1869 - 2009
    Topics: Biology , Medicine
    Notes: [Auszug] Organ xenografts under certain circumstances survive in the presence of anti-graft antibodies and complement, a situation referred to as “accommodation.” We find that the endothelial cells (ECs) in hamster hearts that accommodate themselves in rats express genes, such as A20 and bcl-2, ...
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    Expression of cell-cycle regulators p27 Kip1 and cyclin E, alone and in combination, correlate with survival in young breast cancer patients (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 222-225 
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    Source: Nature Archives 1869 - 2009
    Topics: Biology , Medicine
    Notes: [Auszug] Mutations in certain genes that regulate the cell cycle, such as p16 and p53, are frequently found in human cancers1. However, tumor-specific mutations are uncommon in genes encoding cyclin E and the CDK inhibitor p27Kip1, two cell-cycle regulators that are also thought to contribute to tumor ...
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    Typhoid Mary: Captive to the Public's Health (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 241-242 
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    Source: Nature Archives 1869 - 2009
    Topics: Biology , Medicine
    Notes: [Auszug] by Judith Walzer Leavitt Beacon Press 1996 ISBN: 0-8070-2102-4 $25.00 REVIEWED BY JAMES CURRAN Dean, Rollins School of Public Health, Emory University, 1518 Clifton Rd. NE, Atlanta, GA, 30322 Why don't we treat HIV/AIDS like every other disease? Why shouldn't all carriers of dangerous ...
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    Puzzling over prion partners (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 1322-1322 
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    Source: Nature Archives 1869 - 2009
    Topics: Biology , Medicine
    Notes: [Auszug] To understand what goes wrong in disease, we often need to know how the biological systems usually operate and work backward from there. The study of transmissible spongiform encephalopathies (TSEs)—such as bovine spongiform encephalopathy, Creutzfeldt-Jakob disease and scrapie—is no ...
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    Regeneration in the adult central nervous system: Experimental repair strategies (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 1329-1335 
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    Source: Nature Archives 1869 - 2009
    Topics: Biology , Medicine
    Notes: [Auszug] Genes dictate development of the major cytoarchitectonic plan comprising the central nervous system. During formation, this plan may become disturbed for a variety of reasons, but once development is completed, the principle wiring diagram remains fixed. At the level of a single neuron and ...
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    AMA aims for ethical high road following Sunbeam disaster (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 1183-1183 
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    Source: Nature Archives 1869 - 2009
    Topics: Biology , Medicine
    Notes: [Auszug] In an effort to clean house following its debacle with Sunbeam (Delray Beach, FL), the American Medical Association (AMA) announced the departure of three executives key to the ill-fated endorsement deal. A week later it publicized a plan to gather a panel of nationally known medical ethicists to ...
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    $34 million neuropeptide deal (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 1183-1183 
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    Source: Nature Archives 1869 - 2009
    Topics: Biology , Medicine
    Notes: [Auszug] Neuropeptide research is becoming increasingly valuable, not only in terms of its contribution to medical science but also in terms of hard cash. Warner Lambert recently signed a $34 million deal with New Jersey based Synaptic Pharmaceuticals to “advance research on the central ...
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    DNA Simplified: The Hitchhiker's Guide to DNA (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 1409-1409 
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    Source: Nature Archives 1869 - 2009
    Topics: Biology , Medicine
    Notes: [Auszug] by Daniel H. Farkas AACC Press, Washington, DC, 104 pp. ISBN 0-915274-84-1 Reviewed by Carina Dennis Assistant Editor, Nature Genetics “If you are good in the kitchen, you're good in the laboratory and vice versa”, according to Daniel Farkas, in his latest book, DNA Simplified: The ...
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    Mass spectrometry from miniaturized arrays for full comparative DNA analysis (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 1413-1416 
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    Source: Nature Archives 1869 - 2009
    Topics: Biology , Medicine
    Notes: [Auszug] The wealth of genetic information stemming from the Human Genome Project has broad implications impacting biomedical and pharmaceutical research1. A critical cornerstone is defining the role of gene function as a contributor to etiology of diseases, but making such correlations is complex given the ...
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    Dimers are a girl's best friend (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 1199-1200 
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    Source: Nature Archives 1869 - 2009
    Topics: Biology , Medicine
    Notes: [Auszug] NOT EVERYONE EXPOSED to HIV develops the circulating antibodies that are the hallmark of persistent infection and ultimate immunological collapse. Reaching a better understanding of how some people resist HIV infection should provide valuable clues in the search for an effective vaccine. The ...
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    The UK endorses DIY genetic screening (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 1180-1180 
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    Source: Nature Archives 1869 - 2009
    Topics: Biology , Medicine
    Notes: [Auszug] The UK has become one of the first countries in the world to endorse mail-order genetic screening. But in doing so, the Department of Health has announced a stringent list of requirements that it says it will expect to be met by any company planning to market genetic screening tests directly to the ...
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    A novel protein that participates in nonself discrimination of malignant cells by homologous complement (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 1266-1270 
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    Source: Nature Archives 1869 - 2009
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    Notes: [Auszug] The human complement (C) system protects an individual against substances of nonself origin, including xenografts and microbial pathogens1. Human cells express C-regulatory proteins, CD46 and CD55, thereby circumventing attack by C3, a major effector of C (ref. 2). Nevertheless, certain malignant ...
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    On the market (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 1297-1299 
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    Source: Nature Archives 1869 - 2009
    Topics: Biology , Medicine
    Notes: [Auszug] UNDER THE HAMMER Going, Going … Sold is a new auction service on the World Wide Web specializing in used and refurbished laboratory equipment. This new service, offered by Internet Auctioneers International, is said to offer significant advantages over conventional sales methods for both ...
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    Estrogen accelerates cutaneous wound healing associated with an increase in TGF-β1 levels (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 1209-1215 
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    Source: Nature Archives 1869 - 2009
    Topics: Biology , Medicine
    Notes: [Auszug] The cellular and molecular mechanisms underlying the effects of aging on human cutaneous wound healing are poorly understood, and the possible role of reproductive hormones in this process has never been investigated. We report that aging in healthy females was associated with a reduced rate of ...
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    CCR2 chemokine receptor and AIDS progression (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 1052-1053 
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    Source: Nature Archives 1869 - 2009
    Topics: Biology , Medicine
    Notes: [Auszug] Recently we reported an association between the CCR2-64I chemokine receptor mutation and delayed onset of AIDS1. Michael et al. have now examined the San Francisco Mens Health Study (SFMHS) AIDS cohort for CCR2 genotypes and in this issue of Nature Medicine report that they fail to detect the ...
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    From human rights to the human genome : stopping biopiracy (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 1056-1056 
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    Source: Nature Archives 1869 - 2009
    Topics: Biology , Medicine
    Notes: [Auszug] Genetic research by academic institutions and biotechnology companies around the globe is yielding long-sought-after clues to mutations that could help cure a wide range of diseases. This research is being increasingly conducted on tissue samples from native populations known to have increased ...
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    Osteopetrosis in mice lacking NF-κB1 and NF-κB2 (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 1285-1289 
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    Source: Nature Archives 1869 - 2009
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    Notes: [Auszug] The nfkbl and nfkb2 genes encode closely related products regulating immune and inflammatory responses1–3. Their role during development and differentiation remains unclear. The generation of nfkb1 null mice (p50−/−) resulted in altered immune responses, but had no effect on ...
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    Sex, Race, and Science (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 1292-1293 
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    Source: Nature Archives 1869 - 2009
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    Notes: [Auszug] EUGENICS IN THE DEEP SOUTH by Edward J.Larson Johns Hopkins University Press, Baltimore, $14.95,251 pp. ISBN 0-80185-511-X, 1996 REVIEWED BY JON BECKWITH American Cancer Society Research Professor, Harvard Medical School, Boston, MA 02115 Halfway through reading Edward Larson's ...
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    Next year's Nobels? (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 1051-1051 
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    Source: Nature Archives 1869 - 2009
    Topics: Biology , Medicine
    Notes: [Auszug] The Albert Lasker Medical Research Awards were established in 1946 to celebrate scientists, physicians and public servants whose accomplishments have had a major impact on the treatment or understanding of disease. Nature Medicine is delighted to be publishing commentaries penned by the three 1997 ...
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    Italian mental hospitals closer to closing? (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 1054-1054 
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    Source: Nature Archives 1869 - 2009
    Topics: Biology , Medicine
    Notes: [Auszug] To the editor — As reminded by Nature Medicine1, the threat of central government sanctions have put further pressure on the Italian Regions to close all remaining psychiatric hospitals (PH) as part of the country's bold 1978 plan to revolutionize its care of the mentally ill by releasing ...
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    Gastrogenomic delights: A movable feast (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 1076-1078 
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    Source: Nature Archives 1869 - 2009
    Topics: Biology , Medicine
    Notes: [Auszug] RECENTLY, WE BIOLOGISTS have been treated to a feast of the complete genome sequences of two gut bacteria: Helicobacter pylori reported by Tomb et al.1 in Nature and Escherichia coli reported by Blattner et al.2 in Science. Complete sequences of eight microbes have now been published (Table 1), and ...
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    bFGF and tumor angiogenesis : Back in the limelight? (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 1083-1084 
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    Source: Nature Archives 1869 - 2009
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    Notes: [Auszug] RESEARCH ON TUMOR angiogenesis, which began in earnest 25 years ago, is hot news. This was not the case until relatively recently, so why the big change? The answer can be traced to the discovery and molecular characterization of a diverse family of regulators of angiogenesis, both stimulators and ...
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    GLUT4 heterozygous knockout mice develop muscle insulin resistance and diabetes (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 1096-1101 
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    Source: Nature Archives 1869 - 2009
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    Notes: [Auszug] GLUT4, the insulin-responsive glucose transporter, plays an important role in postprandial glucose disposal. Altered GLUT4 activity is suggested to be one of the factors responsible for decreased glucose uptake in muscle and adipose tissue in obesity and diabetes. To assess the effect of GLUT4 ...
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    Various views on anonymity (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 2-2 
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    Notes: [Auszug] To the editor — It is my observation that the quality of manuscript reviews has decreased dramatically over the past five years. Reviewers with professional or economic conflicts of interest, or even mere personal animus, will say anything to block or to delay publication of important ...
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    Transcytosis of infectious human immunodeficiency virus across a tight human epithelial cell line barrier (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 42-47 
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    Notes: [Auszug] Contact between various epithelial cell lines and HIV chronically infected mononuclear cell lines results in a massive and rapid budding of HIV virions toward the epithelium followed by their internalization into epithelial endosome-like structures. Here it is shown that as early as 30 min utes ...
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    Commissioner Kessler's activist legacy (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 6-7 
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    Notes: [Auszug] David A. Kessler, whose aggressive and controversial six-year tenure as commissioner of the US Food and Drug Administration (FDA) touched the lives of virtually every American consumer, will return to private life in February. He leaves an agency largely restored to its original mission as a ...
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    Inositol prevents folate-resistant neural tube defects in the mouse (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 60-66 
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    Notes: [Auszug] Clinical trials demonstrate that up to 70% of neural tube defects (NTDs) can be prevented by folk acid supplementation in early pregnancy, whereas the remaining NTDs are resistant to folate. Here, we show that a second vitamin, myo-inositol, is capable of significantly reducing the incidence ...
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    In the image and likeness of God: Always? (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 7-7 
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    Notes: [Auszug] “It is not the role of the state to decide who shall live and who shall die.” These words, spoken by Joseph Cardinal Ratzinger (prefect of the Roman Catholic Church's Congregation for the Doctrine of the Faith) opened a remarkable conference held recently in Vatican City. Entitled ...
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    Politicians call halt to “drive-through” mastectomies (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 479-479 
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    Notes: [Auszug] New York has become the first state to ban “drive-through” mastectomy — the surgical treatment of breast cancer as an outpatient procedure. The state law, passed last month, comes ahead of a proposed federal bill that aims to stop this increasing practice in the USA. Media ...
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    Britain plans for research into primary care (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 480-480 
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    Notes: [Auszug] Keen to see the National Health Service deliver what it calls “evidence-based” medicine, the British government has launched a major initiative designed to boost research into primary health care. Its goals range from increasing the effectiveness of nursing care to determining what ...
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    T-cell homeostasis in HIV infection is neither failing nor blind: Modified cell counts reflect an adaptive response of the host (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 486-490 
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    Source: Nature Archives 1869 - 2009
    Topics: Biology , Medicine
    Notes: [Auszug] The decisive pathological processes associated with HIV disease occur in infected lymphoreticular tissues (LT). Because our ability to sample these sites is limited, the blood compartment necessarily serves as a window on these processes. Changes in viral load and in T-cell counts, which occurred ...
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1038/nm0597-486
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  • 98
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    Chromosomal breakpoints hit the spot (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 496-497 
    Details
    ISSN: 1546-170X
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Medicine
    Notes: [Auszug] THE IMPORTANCE OF chromosomal abnormalities in tumor development has been clear since pioneering cytogenetic studies successfully defined recurrent changes in specific types of tumors. The concept that proto-oncogenes or suppressor genes are involved in the various forms of microscopically ...
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1038/nm0597-496
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  • 99
    Electronic Resource
    Electronic Resource
    Neurotrophic factor therapy : Keeping score (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 497-498 
    Details
    ISSN: 1546-170X
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Medicine
    Notes: [Auszug] THE THERAPEUTIC BENEFITS of neurotrophic factors have been proclaimed for several years. Unfortunately, their clinical track record so far has not been impressive. Both ciliary neurotrophic factor (CNTF) and brain-derived neurotrophic factor (BDNF) failed to benefit amyotrophic lateral sclerosis ...
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1038/nm0597-497
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  • 100
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    HIV infection induces changes in CD4 + T-cell phenotype and depletions within the CD4 + T-cell repertoire that are not immediately restored by antiviral or immune-based therapies (1997)
    [s.l.] : Nature Publishing Group
    Nature medicine 3 (1997), S. 533-540 
    Details
    ISSN: 1546-170X
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Medicine
    Notes: [Auszug] Changes in CD4+ T-cell surface marker phenotype and antigen receptor (TCR) repertoire were examined during the course of HIV infection and following therapy. A preferential decline in naive CD4+ T cells was noted as disease progressed. Following protease inhibitor therapy, naive CD4+ T cells ...
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1038/nm0597-533
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