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  • 1
    Electronic Resource
    Electronic Resource
    New members of the Escherichia coliσE regulon identified by a two-plasmid system (2003)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 225 (2003), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: A previously established method, based on a two-plasmid system, was used to identify promoters recognized by RNA polymerase containing the extracytoplasmic stress response sigma factor σE in Escherichia coli. In addition to previously identified rpoE-dependent promoters, 11 new promoters potentially directing the expression of 15 genes were identified that were active only after over-expression of rpoE. The promoters were confirmed and transcriptional start points of the promoters were determined by primer extension analysis and S1-nuclease mapping. All the promoters contained sequences similar to the consensus sequence of rpoE-dependent promoters. The new rpoE-dependent promoters governed expression of genes encoding proteins involved in primary metabolism (fusA, tufA, recR), phospholipid and lipopolysaccharide biosynthesis (psd, lpxP), signal transduction (sixA), proposed inner or outer membrane proteins (bacA, sbmA, smpA, yeaY), and proteins with unknown function (ybaB, yaiW, yiiS, yiiT, yfeY).
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1016/S0378-1097(03)00480-4
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  • 2
    Electronic Resource
    Electronic Resource
    Corrigendum to “Optimization of a two-plasmid system for the identification of promoters recognized by RNA polymerase containing Staphylococcus aureus alternative sigma factor σB” [FEMS Microbiol. Lett. 232 (2004) 173–179] (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 235 (2004), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1574-6968.2004.tb09589.x
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  • 3
    Electronic Resource
    Electronic Resource
    The positions of the sigma-factor genes, whiG and sigF, in the hierarchy controlling the development of spore chains in the aerial hyphae of Streptomyces coelicolor A3(2) (1996)
    Oxford, UK : Blackwell Publishing Ltd
    Molecular microbiology 21 (1996), S. 0 
    Details
    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: whiG and sigF encode RNA polymerase sigma factors required for sporulation in the aerial hyphae of Streptomyces coelicolor. Their expression was analysed during colony development in wild-type and sporulation-defective whi mutant strains. Each gene was transcribed from a single promoter. Unexpectedly, whiG mRNA was present at all time points, including those taken prior to aerial mycelium formation; this suggests that whiG may be regulated post-transcriptionally. Transcription of whiG did not depend upon any of the six known‘early’whi genes required for sporulation septum formation (whiA, B, G, H, /and J), placing it at the top of the hierarchy of whi loci. sigF expression appeared to be regulated at the level of transcription; sigF transcripts were detected transiently when sporulation septa were observed in the aerial hyphae. Transcription of sigF depended upon all six of the early whi genes, including whiG. The sigF promoter does not resemble the consensus sequence established for σWhiG-dependent promoters and EnWhiG did not transcribe from the sigF promoter in vitro. Consequently, the genetic dependence of sigF upon whiG is very likely to be indirect. These results show that there is a hierarchical relationship between sigma factors required for Streptomyces sporulation and also that at least five other genes are involved in this transcriptional network.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1365-2958.1996.tb02567.x
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  • 4
    Electronic Resource
    Electronic Resource
    A new RNA polymerase sigma factor, σF is required for the late stages of morphological differentiation in Streptomyces spp. (1995)
    Osney Mead, Oxford OX2 0EL, UK : Blackwell Scientific Publications
    Molecular microbiology 17 (1995), S. 0 
    Details
    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: A gene (sigF) encoding a new sigma factor was isolated from Streptomyces aureofaciens using a degenerate oligonucleotide probe designed from the GLI(KDNE)A motif lying within the well-conserved region 2.2 of the eubacterial σ70 family. Homologues were present in other Streptomyces spp., and that of the genetically well-studied Streptomyces coelicolor A3(2) was also cloned. The nucleotide sequences of the two sigF genes were determined and shown to encode primary translation products of 287 (S. coelicolor) and 295 (S. aureofaciens) amino acid residues, both showing greatest similarity to σB of Bacillus subtilis. However, while σB is involved in stationary-phase gene expression and in the general stress response in B. subtilis, σF affects morphological differentiation in Streptomyces, Disruption of sigF did not affect vegetative growth but did cause a whi mutant phenotype. Microscopic examination showed that the sigF mutant produced spores that were smaller and deformed compared with those of the wild type, that the spore walls were thinner and sensitive to detergents and that in sigF mutant spores the chromosome failed to condense. σF is proposed to control the late stages of spore development in Streptomyces.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1365-2958.1995.mmi_17010037.x
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  • 5
    Electronic Resource
    Electronic Resource
    RNA polymerase heterogeneity in Streptomyces aureofaciens: characterization by antibody-linked polymerase assay (1991)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 90 (1991), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Abstract Using antibody-linked polymerase assay we studied the polypeptide composition of DNA-dependent RNA polymerase from Streptomyces aureofaciens and immunological cross-reaction with Escherichia coli RNA polymerase. We identified about 25 ‘ALPA-reactive’ polypeptides which are probably involved in the transcriptional apparatus. We demonstrated that β′ and β subunits from S. aureofaciens and E. coli are immunologically related and σ70 (E. coli) shows immunochemical similarity with σ35 (S. aureofaciens). According to the reconstitution of RNA polymerase holoenzyme and antibody-linked polymerase assay we identified sigma factors responsible for recognition of two promoters.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1574-6968.1991.tb05125.x
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  • 6
    Electronic Resource
    Electronic Resource
    Identification of nucleotides critical for activity of the σE-dependent rpoEp3 promoter in Salmonella enterica serovar Typhimurium (2004)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 238 (2004), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: We previously described a two-plasmid system for the identification of promoters recognized by Salmonella enterica serovar Typhimurium (S. Typhimurium) σE. The S. Typhimurium σE-dependent rpoEp3 promoter was active in the E. coli two-plasmid system only after arabinose-induced expression of S. Typhimurium rpoE. In the present study, we have exploited this two-plasmid system for the identification of nucleotides critical for activity of the rpoEp3 promoter. A library of randomly mutated DNA fragments containing the rpoEp3 promoter was cloned upstream of a lacZα reporter gene and screened for activity in the presence of S. Typhimurium σE. The clones exhibiting reduced LacZ activity were sequenced to identify the mutations. The activity of the mutated rpoEp3 promoters were studied further using a luciferase-based promoter-probe plasmid. All of the important nucleotides of the rpoEp3 promoter (in capital) were located in the −35 (ggAActt) and −10 (TctaA) regions. The critical nucleotides were also the most conserved in known σE-dependent promoters. The study also revealed the importance of the 16-bp spacing between −10 and −35 region, as reducing the spacing to 15-bp greatly reduced activity of the promoter. This method should be generally applicable for the identification of important nucleotides in the cognate promoters of other σ factors.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1574-6968.2004.tb09760.x
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  • 7
    Electronic Resource
    Electronic Resource
    A new gene, sigG, encoding a putative alternative sigma factor of Streptomyces coelicolor A3(2) (1999)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 172 (1999), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: An oligonucleotide probe encoding a peptide motif conserved in all sigma factors was used to isolate a new gene, sigG, from a Streptomyces coelicolor A3(2) genomic library. The deduced protein of 263 amino acids with an Mr of 29 422 showed the greatest similarity to the previously identified sporulation sigma factor (σF) of Streptomyces coelicolor, and general stress response sigma factor (σB) of Bacillus subtilis, mostly in domains suggested to be involved in recognition of −10 and −35 promoter regions. Southern-blot hybridization with DNA from several Streptomyces spp. revealed the presence of a similar gene in all strains tested. Disruption of the S. coelicolor sigG gene appeared to have no obvious effect on growth, morphology, differentiation, and production of pigmented antibiotic actinorhodin and undecylprodigiosin.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1574-6968.1999.tb13463.x
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  • 8
    Electronic Resource
    Electronic Resource
    Disruption of a sigma factor gene, sigF, affects an intermediate stage of spore pigment production in Streptomyces aureofaciens (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 153 (1997), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The Streptomyces aureofaciens sigF gene encodes a sigma factor. By integrative transformation, via double cross-over, a stable null mutant of sigF gene was obtained. This mutation appeared to have no obvious effect on vegetative growth, but affected the late stage of spore maturation. Microscopic examination showed that spores were deformed, and spore wall was thinner, compared with the wild-type spores. The spore pigment of sigF mutant was green, compared to wild-type grey-pink spore pigmentation. The plasmid-born wild-type sigF gene complemented the mutation after transformation of the mutant strain.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1574-6968.1997.tb12598.x
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  • 9
    Electronic Resource
    Electronic Resource
    The Brevibacterium flavum sigma factor SigB has a role in the environmental stress response (2002)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 216 (2002), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: We have previously cloned a gene encoding a SigB, a principal-like sigma factor in Brevibacterium flavum, which was induced by several stress conditions. To clarify the in vivo function of this sigma factor, the sigB gene was disrupted by a homologous recombination, replacing the internal essential coding region in B. flavum chromosome by a kanamycin resistance marker gene. This mutation dramatically decreased vegetative growth rates of B. flavum. Studies of the effect of the sigB mutation on growth and viability of the cells under conditions of stress showed that the sigB mutant had increased susceptibility to acid, salt, alcohol, heat and cold stress. The plasmid-born wild-type sigB gene complemented the mutation. Based on the results, we propose that SigB has a role in vegetative growth and in response to various environmental stresses.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1574-6968.2002.tb11418.x
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  • 10
    Electronic Resource
    Electronic Resource
    Activity of the Streptomyces coelicolor stress-response sigma factor σH is regulated by an anti-sigma factor (2002)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 209 (2002), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The alternative sigma factor σH has been shown to play an important role in stress response and morphological differentiation in Streptomyces coelicolor. Its gene, sigH, is located in an operon with the gene encoding proposed anti-sigma factor UshX, and one of the promoters directing expression of the operon is dependent upon sigH. To clarify the function of S. coelicolor UshX, both the σH and UshX proteins were overproduced in Escherichia coli and purified. In an in vitro transcription assay, σH, after complementation with S. coelicolor core RNA polymerase, was able to recognize the sigH-dependent promoter, sigH-P2. This transcription was inhibited by UshX, if it was incubated with σH prior to the addition of the core RNA polymerase. When σH and UshX were incubated and electrophoresed through non-denaturing polyacrylamide gels, they formed a specific complex. These results showed that UshX is a specific anti-sigma factor for σH, and the S. coelicolor sigH operon is directly autoregulated.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1574-6968.2002.tb11136.x
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