Studies of sigma D-dependent functions in Bacillus subtilis

Márquez, L. M.; Helmann, John D.; Ferrari, Eugenio; Parker, Helen M.; Ordal, George; Chamberlin, Michael J.

doi:10.1128/jb.172.6.3435-3443.1990

Cited by 138 publications

(150 citation statements)

References 49 publications

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“…Marquez et al (30) found that the lack of &'P causes a decrease in the amounts of major autolysin (amidase) and glucosaminidase. Therefore, the sin (flaDl) mutation might interfere with transcription from the Pd promoter by depressing expression of sigD.…”

Section: Resultsmentioning

confidence: 99%

“…B. subtilis AD2 [purB his-i A(lppXcwlB)::cat] was constructed as described below and used as a host for promoter probing. Pleiotropic mutants B. subtilis CB100 (sigD::pLM5 trpC2 cat) (kindly provided by Michael J. Chamberlin [30]) and B. subtilis AC334 (purB sin [flaDI lyt-1]) (1) were also used. The sin (flaDI) mutation is a GAT-to-GGT change at codon 84, causing an Asp-to-Gly amino acid substitution in Sin (FlaD) (26).…”

Section: Materuils and Methodsmentioning

confidence: 99%

“…No sequences typical of rho-independent terminators separate these genes, raising the possibility that they constitute an operon. Some of pleiotropic mutations [sigD, div-341, sacU(Hy) (pap, amyB) andflaD (yt)] reduce autolysin (amidase and/or glucosaminidase) production (6,9,30,41,47,50). The sigD and div-341 mutations are identical to the flaB and secA mutations, respectively (30,42).…”

mentioning

confidence: 99%

“…Some of pleiotropic mutations [sigD, div-341, sacU(Hy) (pap, amyB) andflaD (yt)] reduce autolysin (amidase and/or glucosaminidase) production (6,9,30,41,47,50). The sigD and div-341 mutations are identical to the flaB and secA mutations, respectively (30,42). The sacU(Hy) mutations, which map to adjacent genes (degU and degS), cause an increase in the production of degradative enzymes, poor development of competence, and lack of flagella (loss of motility) (8,33,37).…”

mentioning

confidence: 99%

“…The product of this gene is not a major autolysin but an N-acetylmuramoyl-L-alanine amidase (10, 22) that is highly active on spore cortex peptidoglycans (10). Recently, we cloned and sequenced the major autolysin gene, cwlB, a mutant deficient in which is extremely resistant to cell lysis even after prolonged incubation (25 (6,9,30,41,47,50). The sigD and div-341 mutations are identical to the flaB and secA mutations, respectively (30, 42).…”

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confidence: 99%

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High-level transcription of the major Bacillus subtilis autolysin operon depends on expression of the sigma D gene and is affected by a sin (flaD) mutation

Kuroda

Sekiguchi

1993

J Bacteriol

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Transcription of the major BaciUlus subtilis autolysin gene (cwlB) was investigated. Deletion of the region upstream of the gene cluster 1ppX-cwbA-cwlB led to a loss of promoter activity. Primer extension analysis suggested that the cwlB operon is transcribed by Eo&E and Eo'A, the former transcripts being predominants at the exponential growth phase. Expression of the lppX-lacZ fusion gene was reduced by about 90% in a sigD-null mutant. A sin (flaDi) mutation caused a severe defect in transcription of the lppX-cwbA-cwlB operon. The sin (flaDi) mutation also reduced expression of a sigD-lacZ fusion gene constructed in the B. subtilis chromosome. Since the sigD-null mutant exhibits motility and autolysin deficiencies and filamentation, similar phenotypes in the sin (faDI) mutant may be caused by reduction in expression of the o'r protein.Peptidoglycan hydrolases are believed to be involved in important biological processes such as cell wall turnover (4,7,18,51) and cell separation (7,9,39) and to act as pacemaker and spacemaker enzymes for cell growth (51). Some of the peptidoglycan hydrolases can act as autolysins, thereby appearing to be potential suicide enzymes. This life and death dichotomy of function demands efficient and strict regulation of peptidoglycan hydrolase activity (18).Bacillus subtilis produces two autolysins, N-acetylmuramoyl-L-alanine amidase (the major autolysin) and endo-p-N-acetylglucosaminidase (glucosaminidase) (16,40). During sporulation, two other cell wall hydrolases, sporulationspecific amidase (s-amidase) and -y-D-glutamyl-L-mesodiaminopimelyl endopeptidase, are formed; the hydrolases are thought to be associated with the formation of cortex peptidoglycans (13). Some B. subtilis strains (YT-25 and K-77) produce extracellular endo-,B-N-acetylmuramidases (21, 34, 35). We previously reported the cloning of the cell wall hydrolase gene, cwlA (24). The product of this gene is not a major autolysin but an N-acetylmuramoyl-L-alanine amidase (10, 22) that is highly active on spore cortex peptidoglycans (10). Recently, we cloned and sequenced the major autolysin gene, cwlB, a mutant deficient in which is extremely resistant to cell lysis even after prolonged incubation (25 (6,9,30,41,47,50). The sigD and div-341 mutations are identical to the flaB and secA mutations, respectively (30, 42). The sacU(Hy) mutations, which map to adjacent genes (degU and degS), cause an increase in the production of degradative enzymes, poor development of competence, and lack of flagella (loss of motility) (8,33,37). The flaD (lyt) mutation causes similar * Corresponding author.phenotypes (an increase in alkaline protease, filamentous cell morphology, poor development of competence, and lack of flagella) (2, 36). Recently, we demonstrated that the flaD (lyt) gene is identical to the sin (sporulation inhibition) gene (11,45,46). Smith and colleagues found that Sin is a 14-kDa dual-function DNA-binding protein which has a negative function in sporulation (spoIL4, spoIIF, and spoIIG) and in aprE (major alkaline pr...

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Section: Resultsmentioning

confidence: 99%

Section: Materuils and Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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High-level transcription of the major Bacillus subtilis autolysin operon depends on expression of the sigma D gene and is affected by a sin (flaD) mutation

Kuroda

Sekiguchi

1993

J Bacteriol

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Two‐dimensional polyacrylamide gel electrophoresis of proteins synthesized during early germination of Bacillus subtilis 168 in the presence of actinomycin D

Hirano

Matsuda

Kameyama

1991

J. Basic Microbiol.

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Two-dimensional gel electrophoresis was used to demonstrate the synthesis of approximately 65 [35S]-methionine-labelled soluble proteins between 0 and 10 min after the start of germination, of approximately 210 proteins at 10-20 min, and of approximately 260 proteins during vegetative growth of Bacillus subtilis. When actinomycin D and [35S]-methionine were administered at the onset of germination and the proteins synthesized during the subsequent 15 min were analyzed, two proteins were detected, and were designated protein I and protein II. Immunoblot analysis with an antiserum raised against RNA polymerase from Escherichia coli demonstrated that protein II corresponded to the sigma A factor of Bacillus subtilis. Thus, the sigma A factor is synthesized during early germination of Bacillus subtilis in the presence of actinomycin D.

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The dynamic protein partnership of RNA polymerase inBacillus subtilis

Delumeau¹,

Lecointe²,

Muntel³

et al. 2011

Proteomics

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In prokaryotes, transcription results from the activity of a 400 kDa RNA polymerase (RNAP) protein complex composed of at least five subunits (2α, β, β', ω). To ensure adequate responses to changing environmental cues, RNAP activity is tightly controlled by means of interacting regulatory proteins. Here, we report the affinity-purification of the Bacillus subtilis RNAP complexes from cells in different growth states and stress conditions, and the quantitative assessment by mass spectrometry of the dynamic changes in the composition of the RNAP complex. The stoichiometry of RNA polymerase was determined by a comparison of two mass spectrometry-based quantification methods: a label-based and a label-free method. The validated label-free method was then used to quantify the proteins associated with RNAP. The levels of sigma factors bound to RNAP varied during growth and exposure to stress. Elongation factors, helicases such as HelD and PcrA, and novel unknown proteins were also associated with RNAP complexes. The content in 6S RNAs of purified RNAP complexes increased at the onset of the stationary phase. These quantitative variations in the protein and RNA composition of the RNAP complexes well correlate with the known physiology of B. subtilis cells under different conditions.

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Studies of sigma D-dependent functions in Bacillus subtilis

Cited by 138 publications

References 49 publications

High-level transcription of the major Bacillus subtilis autolysin operon depends on expression of the sigma D gene and is affected by a sin (flaD) mutation

High-level transcription of the major Bacillus subtilis autolysin operon depends on expression of the sigma D gene and is affected by a sin (flaD) mutation

Two‐dimensional polyacrylamide gel electrophoresis of proteins synthesized during early germination of Bacillus subtilis 168 in the presence of actinomycin D

The dynamic protein partnership of RNA polymerase inBacillus subtilis

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