Cloning and characterization of the regulatory Bacillus subtilis competence genes comA and comB

Guillén, Nancy; Weinrauch, Yvette; Dubnau, David

doi:10.1128/jb.171.10.5354-5361.1989

Cited by 55 publications

(54 citation statements)

References 27 publications

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“…The ComP-ComA system also controls expression of the late competence genes comC and comG. Expression of comC and comG is stimulated by nitrogen limitation in the presence of glucose and repressed by glutamine (2,10,28,47). It has been suggested that the ComP-ComA system is involved in this metabolic control (47).…”

Section: Resultsmentioning

confidence: 99%

DegS-DegU and ComP-ComA modulator-effector pairs control expression of the Bacillus subtilis pleiotropic regulatory gene degQ

et al. 1991

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Production of a class of both secreted and intracellular degradative enzymes in Bacillus subtilis is regulated at the transcriptional level by a signal transduction pathway which includes the DegS-DegU two-component system and at least two additional regulatory genes, degQ and degR, encoding polypeptides of 46 and 60 amino acids, respectively. Expression of degQ was shown to be controlled by DegS-DegU. This Production of a class of degradative enzymes in Bacillus subtilis, including an intracellular protease and several secreted enzymes (levansucrase, alkaline and metalloproteases, a-amylase, 3-glucanase[s], and xylanase) (3,5,11,29), is controlled at the transcriptional level by the DegSDegU two-component system and by at least two additional regulatory genes, degQ and degR, which encode small polypeptides of 46 and 60 amino acids, respectively (3,25,42,48,49).Although the two-component system is required for degradative enzyme production, the products of degQ and degR appear to be dispensable (48,49). Mutations were identified in both degS and degU leading to either deficiency of degradative enzyme production or a pleiotropic (Hy) phenotype, which includes hyperproduction of degradative enzymes, ability to sporulate in the presence of glucose, decreased genetic competence, and loss of flagella (6,12,17,23).Since degradative enzyme production requires both the DegS protein kinase and the DegU effector, we postulated that the phosphorylated form of DegU may be necessary for this process. On the other hand, we postulate that the nonphosphorylated form of DegU may be required for competence (23, 28; this report). This hypothesis is supported by the following observations: (i) the degUJ46 mutation abolishing the putative site of phosphorylation of the DegU effector did not abolish competence (23), and (ii) deletion of the degS gene did not strongly reduce competence (this report).The degQ36 mutation, a single base change at position -10 leading to overexpression of the degQ gene, gave a Hy phenotype similar to that of the degS(Hy) and degU(Hy) * Corresponding author. mutations (3, 48). The increase of degradative enzyme production due to overproduction of DegQ is, however, strictly dependent upon the presence of a functional DegSDegU two-component system (3; this report).We previously reported that expression of degQ was decreased in strains carrying either a deletion of degS and degU or a degU32(Hy) mutation (23). In this report, we show that degQ expression is regulated by a second twocomponent system controlling competence in B. subtilis: 47). Several findings had suggested that this could be the case: (i) DegS-DegU and ComP-ComA show strong amino acid sequence similarities, (ii) both of these two-component systems control the expression of competence, and (iii) the degQ, comP, and comA genes are located at adjacent positions on the B. subtilis chromosome (47).The DegS, DegU, and DegQ proteins are produced at different times during growth. The degS-degU operon is expressed throughout the exponential growth pha...

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Section: Resultsmentioning

confidence: 99%

DegS-DegU and ComP-ComA modulator-effector pairs control expression of the Bacillus subtilis pleiotropic regulatory gene degQ

et al. 1991

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“…Exonuclease III footprinting was done by forming the Sin-DNA probe complexes in 20 p1l of binding buffer, as described above, in the gel retardation assay except that binding buffer also contained 10 5 [lI of 3 M sodium acetate (pH 7.0), and 120 ,u of cold ethanol, and the DNA was precipitated by centrifugation as described above. The samples were analyzed on 6% sequencing gel as described above.…”

Section: Methodsmentioning

confidence: 99%

“…An inactivation of the sin gene (loss of function) results in loss of competence and motility. The effect on competence has now been demonstrated at the level of late com gene expression (10). In addition, the loss of function results in filamentous growth and a deficiency in autolysin production (8,24).…”

mentioning

confidence: 99%

The Bacillus subtilis sin gene, a regulator of alternate developmental processes, codes for a DNA-binding protein

Gaur¹,

Oppenheim²,

Smith³

1991

J Bacteriol

109

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The sin gene of Bacillus subtilis encodes a dual-function regulatory protein, Sin, which is a negative as well as a positive regulator of alternate developmental processes that are induced at the end of vegetative growth in response to nutrient depletion. Sin has been purified to homogeneity by using a simple two-step procedure. It was found to bind to the developmentally regulated aprE (alkaline protease) gene at two sites in vitro. The stronger Sin-binding site (SBS-1) is located more than 200 bp upstream from the transcription start site. It is required for Sin repression of aprE expression in vivo, as strains bearing SBS-1 deletions were not affected by the sin gene. The second, weaker Sin-binding site lies on a DNA fragment that contains the aprE promoter. Results of DNase I, exonuclease III, and dimethyl sulfate footprinting analysis of SBS-1 suggested that Sin binding involves two adjacent binding sites which appear to contain two different partial dyad symmetries. An analysis of the predicted amino acid sequence of Sin revealed a potential leucine zipper protein dimerization motif which is flanked by two helix-turn-helix motifs that could be involved in recognizing two different dyad symmetries.

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“…The comA gene product is required for the expression of a srfA::Tn917lac fusion (21) as well as the com genes that function in the later stages of competence development (2,5). This observation suggests that surfactin production and competence development are regulated by a common signal transduction pathway.…”

mentioning

confidence: 99%

srfA is an operon required for surfactin production, competence development, and efficient sporulation in Bacillus subtilis

et al. 1991

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The srfA locus of BaciUlus subtilis is defined by a transposon Tn917 insertion and is required for productpon of the peptide secondary metabolite surfactin. The srfA locus was isolated by cloning the DNA flanking srfA::Tn917 insertions followed by chromosome walking. The cloned region is an operon of oyer 25 kb which covers the transcription initiation region but not the intact 3' end of srfA. csh-293, which was previously identified as a Tn9171ac mutation that impairs competence development and causes a condifoWl defect in sporulation, was known to be located in the vicinity of the srfA locus within the B. subtlis genome. The csh-293::Tn9171ac mutation was discovered to cause a defect in surfactin production and was shown to be located in the srfA operon by its cotransformation with srfA mutations and' by Southern hybridization analysis. Insertion mutations in srfA, created by the chromosomal integration of plasmids bearing overlapping srfA DNA fragments, were ex for their effects on surfactin production, competence, and sporulation. AB three processes were found to require the intact 5' half of the srfA operon, whereas the 3' half of srfA was found to be required for sporulation and surfactin production but not competence. These experiments show that srfA gene products function in B. subtilis cell specialization and differentiation.

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Cloning and characterization of the regulatory Bacillus subtilis competence genes comA and comB

Cited by 55 publications

References 27 publications

DegS-DegU and ComP-ComA modulator-effector pairs control expression of the Bacillus subtilis pleiotropic regulatory gene degQ

DegS-DegU and ComP-ComA modulator-effector pairs control expression of the Bacillus subtilis pleiotropic regulatory gene degQ

The Bacillus subtilis sin gene, a regulator of alternate developmental processes, codes for a DNA-binding protein

srfA is an operon required for surfactin production, competence development, and efficient sporulation in Bacillus subtilis

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