Heterogeneity of RNA polymerase in Bacillus subtilis: evidence for an additional sigma factor in vegetative cells.

Wiggs, Janey L.; Gilman, Michael; Chamberlin, Michael J.

doi:10.1073/pnas.78.5.2762

Cited by 78 publications

(46 citation statements)

References 38 publications

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“…Five Fig. 1 (58). This PstI fragment is also present as part of the cloned insert of the lambda phage X1-26, independently isolated on the basis of possession of promoter PD-6, the B. subtilis flagellin gene (hag) promoter (20,46).…”

Section: Materuils and Methodsmentioning

confidence: 99%

“…1. The original plasmid which contains promoter PD-1 is pCD4136; it consists of a 2.5-kb PstI fragment of B. subtilis DNA cloned in the vector pHV14 (58). A 1.1-kb PstI-EcoRI fragment from pCD4136 had been subcloned previously into pBR322 and pJH101 to yield pMG102 and pEAK1, respectively (21,30,54).…”

Section: Materuils and Methodsmentioning

confidence: 99%

“…This approach has led previously to identification of the B. subtilis hag gene, encoding the flagellar filament structural protein flagellin, and the cotranscribed motA and motB genes, which are required for the function of the flagellum and motility of the cell (46,47). In this report, we continue using this approach towards the characterization of the transcription unit controlled by one of the first cD-dependent promoters identified, PD-1 (58). We have found that the PD-1 operon contains structural and regulatory flagellar genes, including the B. subtilis homologs of Salmonella typhimurium flgM and flgK.…”

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confidence: 98%

“…Gillen This work a All of the strains except CB100 are derived from the parent CB25 (originally JH642; Fla' Mot' Che+ Lyt+ trpC2pheA1) and were constructed as described in the text. CB100 is derived from I168 (trpC2) as described in reference 27. subtilis DNA fragments each known to contain a strong CD promoter, denoted a PD (20,21,58). This approach has led previously to identification of the B. subtilis hag gene, encoding the flagellar filament structural protein flagellin, and the cotranscribed motA and motB genes, which are required for the function of the flagellum and motility of the cell (46,47).…”

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confidence: 99%

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Identification of flagellar synthesis regulatory and structural genes in a sigma D-dependent operon of Bacillus subtilis

1994

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The SD form of RNA polymerase from BaciUlus subtilis has been shown previously to direct the synthesis of several transcription units bearing genes for flagellin, motility proteins, and autolysins. In this report, we describe an operon of genes transcribed from the orD-dependent promoter PD-We have identified three complete open reading frames and one partial one downstream of this promoter, immediately upstream is the previously identified comF locus. The PD-1 operon encodes the presumptive B. subtilis homologs of two Salmonella typhimurium late flagellar genes,flgM andflgK. Also present in this operon are two genes of unknown function, orfl39 and orfl60, whose products show similarities to the eukaryotic cytoskeletal proteins myosin and vimentin, respectively. orfl39 and orfl60 may encode proteins that form extended a-helical secondary structures and coiled-coil quaternary structures which may be filamentous components of the gram-positive bacterial flagellum. We have characterized the B. subtilisflgM gene further by constructing an in-frame deletion mutation, flgMA80, and creating strains of B. subtilis in which this allele has replaced the wild-type copy. By primer extension analysis of cellular RNA, we have shown that the flgMA80 mutation relieves the block to transcription of two other yD-dependent operons imposed by an unlinked mutation in a gene directing early flagellar synthesis. We conclude that, as in the case of S. typhimurium, early flagellar synthesis in B. subtilis is coupled to late flagellar synthesis through repression of (FD-dependent transcription by theflgM gene product.The specificity of promoter recognition by RNA polymerase, often a key step in bacterial gene expression, is determined by proteins known as sigma (cr) factors that confer recognition of distinct promoter sequences. The gram-positive bacterium Bacillus subtilis possesses many alternative sigma factors and employs them in the temporal and developmental regulation of specific sets of genes, or regulons. B. subtilis cr' is a secondary sigma factor that controls a regulon of genes expressed in exponential and early postexponential growth (19, 20, 42, 46; see reference 26 for a review). Mutations in the structural gene for cD lead to defects in flagellation, motility, and/or chemotaxis functions (28,42). crD is the structural homolog of the enteric flagellar-specific sigma factor crF, and each factor appears to control the transcription of a similar regulon of genes required for flagellar assembly (fla), motility (mot), and potentially chemotaxis (che) functions (for a review, see reference 50). Sigma factors having this same promoter specificity are implicated in flagellar gene expression in a wide variety of bacterial species (26) and appear to form a family of sigma factors with related promoter specificities.The gene expression hierarchy that controls flagellar morphogenesis in the enteric bacteria, the fla/che/mot regulon, has been depicted primarily as a linear pathway (17, 37; see reference 40 for a review). The expressi...

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Section: Materuils and Methodsmentioning

confidence: 99%

Section: Materuils and Methodsmentioning

confidence: 99%

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confidence: 98%

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confidence: 99%

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Identification of flagellar synthesis regulatory and structural genes in a sigma D-dependent operon of Bacillus subtilis

1994

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“…In addition to o'r -containing RNA polymerase (Eor43), vegetative and early stationary-phase cells contain at least three minor forms of RNA polymerase holoenzyme. These holoenzymes contain oa species designated &r37 (3), o-32 (4), and a28 (5), which direct the transcription initiation of distinct sets of cognate promoters. A fifth holoenzyme form containing a o species designated a,29 (6), which is encoded by the sporulation gene spolIG (7), is present exclusively in sporulating cells and recognizes the promoter sites of genes that are switched on at an early to intermediate stage of sporulation (8).…”

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Gene encoding the sigma 37 species of RNA polymerase sigma factor from Bacillus subtilis.

Binnie¹,

Lampe²,

Losick³

1986

Proc. Natl. Acad. Sci. U.S.A.

112

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ABSTRACTaJ37 is a minor species of RNA polymerase cr factor found in the Gram-positive bacterium Bacillus subtilis. 37 governs the transcription in vitro of genes that are turned on at an early stage in spore formation, as well as other genes that are switched on at the end of the exponential phase of growth but that are not under sporulation control. To study the role of O37 in B. subtilis gene expression, we have cloned the gene for this minor species of (F factor in Escherichia coli by using as a hybridization probe a synthetic oligonucleotide that was designed on the basis of the NH2-terminal amino acid sequence of o'37 protein. We determined the nucleotide sequence of the entire o.37 gene, which was found to encode a 262-amino acid residue polypeptide of 29.9 kDa. The predicted amino acid sequence of a37 showed significant homology to that of other (r proteins in a region that has been proposed to be the site ofbinding of these factors to core RNA polymerase. holoenzyme forms in vitro, is induced at the end of the exponential phase of growth but under nutritional conditions that do not support spore formation (38). Unlike spoVG transcription, ctc RNA synthesis is not dependent on the products ofspoO genes but is instead stimulated by mutations in these regulatory genes. Studies on the interaction of E&37 with the ctc and spoVG promoters indicate that promoter recognition by this minor holoenzyme form is determined by contacts with conserved bases in the "-35" and "-10" regions of its cognate promoters (refs. 19 and 21-23; P. Zuber, J. Healy, and R.L., unpublished results).Although much has been learned about the interaction of Eoa37 with spoVG and ctc from in vitro studies, it has not yet been possible to assess the contribution of this minor holoenzyme form to the expression and differential regulation of these genes in vivo or to investigate the general role of Eo-37 in the overall pattern of B. subtilis gene expression.As a way of approaching this problem, we have cloned in Escherichia coli the structural gene for a37 and we have determined the location of its coding sequence on the B. subtilis chromosome. We show that the predicted product of the 37 gene, which we designate sigB, contains a region of significant homology to other B. subtilis and E. coli a factors and is evidently not essential for growth or sporulation, as judged by the construction of a sigB insertion mutation. MATERIALS AND METHODSBacterial Strains and Plasmids. Cloning of DNA from B. subtilis strain 168 (trpC2) was carried out in E. coli strains MM294 and HB101 using the plasmid vectors pBR322 and pJH101 (24). Mapping experiments were carried out in B. subtilis strain QB928 (aroI, purB33, dal, and trpC2; see ref. 25).Abbreviations: kb, kilobase(s); bp, base pair(s). 5943The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.

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Regulation of Bacterial Gene Expression

Reznikoff

1989

Bacteria in Nature

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Heterogeneity of RNA polymerase in Bacillus subtilis: evidence for an additional sigma factor in vegetative cells.

Cited by 78 publications

References 38 publications

Identification of flagellar synthesis regulatory and structural genes in a sigma D-dependent operon of Bacillus subtilis

Identification of flagellar synthesis regulatory and structural genes in a sigma D-dependent operon of Bacillus subtilis

Gene encoding the sigma 37 species of RNA polymerase sigma factor from Bacillus subtilis.

Regulation of Bacterial Gene Expression

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