Identification of transposon-inserted mutations including rnpB::Tn that abolished glucose induction of sigX encoding extracytoplasmic function-sigma factor in Bacillus subtilis

2021 ◽  
Author(s):  
Mitsuo Ogura
Keyword(s):  
Bacillus Subtilis ◽  
Sigma Factor ◽  
Stringent Response ◽  
Rnase P ◽  
Glucose Induction ◽  
Extracytoplasmic Function Sigma Factor

Abstract We investigated the regulators of the glucose induction (GI) of the ECF-sigma genes sigX/M. During further screening of transposon-inserted mutants, we identified several regulators including an RNA component of RNase P (rnpB), which is required for tRNA maturation. A depletion of rnpB is known to trigger the stringent response. We showed evidence that the stringent response inhibited GI of sigX/M.

scholarly journals Regulation of the Bacillus subtilis Extracytoplasmic Function Protein σY and Its Target Promoters

2003 ◽  
Vol 185 (16) ◽  
pp. 4883-4890 ◽  
Author(s):  
Min Cao ◽  
Letal Salzberg ◽  
Ching Sung Tsai ◽  
Thorsten Mascher ◽  
Carla Bonilla ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Sigma Factor ◽  
Negative Regulation ◽  
Wild Type ◽  
Induced Mutations ◽  
Mutant Strains ◽  
Elevated Expression ◽  
Direct Target ◽  
Extracytoplasmic Function Sigma Factor ◽  
Target Promoters

ABSTRACT The Bacillus subtilis extracytoplasmic function sigma factor σY is of unknown function. We demonstrate that the sigY operon is expressed from an autoregulatory promoter site, PY. We selected for transposon-induced mutations that upregulate PY transcription in an attempt to identify genes involved in σY regulation. The resulting insertions disrupted yxlC, the gene immediately downstream of sigY. However, the phenotype of the yxlC::Tn10 insertion was due to polarity on the downstream genes of the sigY operon; a nonpolar insertion in yxlC did not lead to derepression of PY. Further analyses revealed that both yxlD and yxlE encoded proteins important for the negative regulation of σY activity. A comparison of the transcriptomes of wild-type and yxlC::Tn10 mutant strains revealed elevated expression of several operons. However, only one additional gene, ybgB, was unambiguously identified as a direct target for σY. This was supported by analysis of direct targets for σY transcription with whole-genome runoff transcription followed by macroarray analysis.

scholarly journals SigM, an Extracytoplasmic Function Sigma Factor of Bacillus subtilis, Is Activated in Response to Cell Wall Antibiotics, Ethanol, Heat, Acid, and Superoxide Stress

2003 ◽  
Vol 185 (12) ◽  
pp. 3491-3498 ◽  
Author(s):  
Penny D. Thackray ◽  
Anne Moir
Keyword(s):  
Cell Wall ◽  
Bacillus Subtilis ◽  
Exponential Growth ◽  
Sigma Factor ◽  
Promoter Region ◽  
Normal Cell ◽  
Tween 20 ◽  
Exponential Growth Phase ◽  
Extracytoplasmic Function Sigma Factor ◽  
Triton X

ABSTRACT The extracytoplasmic function sigma M of Bacillus subtilis is required for normal cell growth under salt stress. It is expressed maximally during exponential growth and is further induced by the addition of 0.7 M NaCl. The promoter region of the sigM operon contains two promoters; one (PA) is sigma A dependent, and the other (PM) is sigma M dependent. These have been placed separately at the amy locus, directing expression of a lacZ reporter gene. Only the PM fusion responded to salt induction. This promoter, which was responsive to the level of active sigma M in the cell, was also induced by 5% ethanol, by vancomycin, bacitracin, or phosphomycin (inhibitors of cell wall biosynthesis; 2 μg per ml), and by heat shock of 50°C for 10 min. It was very strongly induced by acid (pH 4.3) and 80 μM paraquat, but after a 15- to 30-min delay. There was no induction by alkali (pH 9), 5 mM H2O2, the detergents 0.1% Triton X-100 and 0.1% Tween 20, or 50 μM monensin. In addition to their reduced tolerance to salt, null mutants of sigM were unable to grow at pH 4.3 and lysed after exposure to 5% ethanol. Genes regulated by SigM were also tested for their response to pH 4.3, 5% ethanol, and 2 μg of vancomycin per ml. Expression of the genes may have been activated by increased levels of sigma M, but at least some were also subject to additional controls, as they responded to one type of stress but not another. Expression of yrhJ, which encodes a cytochrome P450/NADPH reductase, was induced in response to acid and vancomycin. yraA expression was acid, ethanol, and vancomycin induced, whereas yjbD showed only ethanol induction. YraA protein was extremely important to acid survival—a mutation in yraA, like a sigM mutation, resulted in the failure of B. subtilis to grow at pH 4.3. Sigma M is therefore involved in maintaining membrane and cell wall integrity in response to several different stresses in exponential growth phase and is activated by such stresses.

scholarly journals SigM-Responsive Genes of Bacillus subtilis and Their Promoters

2007 ◽  
Vol 189 (12) ◽  
pp. 4534-4538 ◽  
Author(s):  
Adrian J. Jervis ◽  
Penny D. Thackray ◽  
Chris W. Houston ◽  
Malcolm J. Horsburgh ◽  
Anne Moir
Keyword(s):  
Bacillus Subtilis ◽  
Sigma Factor ◽  
Rapid Amplification ◽  
Extracytoplasmic Function Sigma Factor

ABSTRACT Promoters of nine Bacillus subtilis genes (bcrC, yacK, ydaH, yfnI, yjbD, ypbG, ypuA, yraA, and ysxA), all responsive to artificially induced increases in the stress-responsive extracytoplasmic function sigma factor, SigM, were mapped by rapid amplification of cDNA ends-PCR. The resulting promoter consensus suggests that overlapping control by SigX or SigW is common.

scholarly journals DegU-phosphate activates expression of the anti-sigma factor FlgM in Bacillus subtilis

2011 ◽  
Vol 81 (4) ◽  
pp. 1092-1108 ◽  
Author(s):  
Yi-Huang Hsueh ◽  
Loralyn M. Cozy ◽  
Lok-To Sham ◽  
Rebecca A. Calvo ◽  
Alina D. Gutu ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Sigma Factor
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