Effects on growth and sporulation of inactivation of a Bacillus subtilis gene (ctc) transcribed in vitro by minor vegetative cell RNA polymerases (E-σ37, E-σ32)

Truitt, Christine L.; Weaver, Elizabeth A.; Haldenwang, W G

doi:10.1007/bf00322460

Cited by 23 publications

(23 citation statements)

References 21 publications

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“…After electrophoretic transfer to nitrocellulose and blocking of the nitrocellulose with BLOTTO, the immobilized protein bands were probed with anti-u'3, anti-RsbW, or anti-RsbV monoclonal antibodies. The anti-RsbV and anti-RsbW antibodies were prepared and analyzed for specificity as was previously described for the anti-u'3 monoclonal antibody (4 gene confers a subtle temperature-dependent phenotype on strains which carry it (2,28). This observation implies that although crB is not essential for cell viability during heat shock, at least a part of its regulon might still participate in the cell's response to thermal stress.…”

Section: Methodsmentioning

confidence: 99%

“…The temperature-sensitive phenotype of the ctc mutant initially suggested that (rB could be the B. subtilis counterpart of &32; however, additional experiments did not support this model. In contrast to E. coli, in which a-32 is critical to cell viability at elevated temperatures (30), B. subtilis strains with null mutations in sigB are no more temperature sensitive then their wild-type counterparts (28). The dispensability of ao' to B. subtilis survival at elevated temperatures argues that a-B isn't equivalent to a;-2 and that if it plays a role during heat shock, this role is not critical.…”

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confidence: 99%

“…The roles of ctc and csbA, like that of itself, are undefined. Neither gene is essential for growth or sporulation (8,28). The only phenotype observed after a loss of one of these genes is temperature-sensitive oligosporageny which occurred upon disruption of ctc by an integrating plasmid (pAK-6/p1949) (28).…”

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confidence: 99%

“…Neither gene is essential for growth or sporulation (8,28). The only phenotype observed after a loss of one of these genes is temperature-sensitive oligosporageny which occurred upon disruption of ctc by an integrating plasmid (pAK-6/p1949) (28). Placement of this ctc null mutation into a strain that carried a mutation in the closely linked but distinct spoVC gene (spoVC285) resulted in a growth as well as a sporulation defect at elevated temperatures.…”

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confidence: 99%

“…Placement of this ctc null mutation into a strain that carried a mutation in the closely linked but distinct spoVC gene (spoVC285) resulted in a growth as well as a sporulation defect at elevated temperatures. Growth of the ctc::pAK-6/p1949 spoVC285 strain was transiently inhibited when exponential cultures were shifted from 37 to 48°C (28).…”

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The sigma B-dependent promoter of the Bacillus subtilis sigB operon is induced by heat shock

1993

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oE, a secondary sigma factor of Bacillus subtilis, was found to increase 5-to 10-fold when cultures were shifted from 37 to 48°C. Western blot (immunoblot) analyses, in which monoclonal antibodies specific for the sigB operon products RsbV, RsbW, and crB were used to probe extracts from wild-type and mutant B. subtilis strains, revealed that all three proteins increased coordinately after heat shock and that this increase was dependent on &rB but not RsbV, a positive regulator normally essential for &3-dependent sigB expression.Nuclease protection experiments of RNA synthesized after heat shock supported the notion that the shift to 48°C enhanced transcription from the sigB operon's o'B-dependent promoter. The level of mRNA initiating at the &B-dependent ctc promoter was also seen to increase approximately 5-to 10-fold after heat shock.Pulse-labeling of the proteins synthesized after a shift to 48°C demonstrated that sigB wild-type and mutant strains produced the maijor heat-inducible proteins in similar amounts; however, at least seven additional proteins were present after the temperature shift in the wild-type strain but absent in the sigB null mutant.Thus, although cr' is not required for the expression of essential heat shock genes, it is activated by heat shock to elevate its own synthesis and possibly the synthesis of several other heat-inducible proteins.a3 is a minor abundance sigma factor of Bacillus subtilis that is found associated with the RNA polymerase from vegetatively growing bacteria but which disappears from the extractable RNA polymerase population by the second hour after the onset of sporulation (12). Although ae3 was the first secondary sigma factor to be discovered in bacteria, its physiological role remains obscure (6, 9, 13). Null mutations in the aW structural gene (sigB) have no obvious effect on vegetative growth or sporulation .6, 9).Even though the function of a' is not yet known, it is a potent regulatory protein with an elaborate mechanism for keeping its activity in check. sigB is the third gene in a four-gene operon with the other genes, rsbV, rsbW, and rsbX (formerly called orfV, orfW, and orfX, respectively), involved in controlling ar3 activity (3, 4, 7). Previous work suggested that the three gene products operate in a pathway of negative control, with RsbW being the primary inhibitor of ar3 activity and the other two gene products (RsbV and RsbX) functioning upstream of RsbW in the regulatory pathway (3, 7). Mutations which reduce RsbW's inhibitory effects severely impair the growth of strains which carry them and readily give rise to suppressor mutations which reduce or eliminate the activity of in these strains (2,3,7,15). In addition to the sigB operon itself (3, 17), two genes (ctc and csbA) which depend on ae for a significant part of their expression have been identified (8,14,16 genes is temperature-sensitive oligosporageny which occurred upon disruption of ctc by an integrating plasmid (pAK-6/p1949) (28). Placement of this ctc null mutation into a strain that carri...

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Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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The sigma B-dependent promoter of the Bacillus subtilis sigB operon is induced by heat shock

1993

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Intracellular reactive oxygen species mediate suppression of sporulation in Bacillus subtilis under shear stress

Sahoo

Rao

Suresh

et al. 2004

Biotech & Bioengineering

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Sporulation is an important cellular response to stress that is also significant from a bioreactor operation viewpoint. While sporulating organisms are known to show an enhanced sporulation response under several stress situations, the sporulation response to shear stress has not been investigated thus far. Such a study could be of interest since shear stress, to a greater or lesser degree, is always present in bioreactor operation. In this article, we investigate the sporulation extents of the Gram-positive bacteria Bacillus subtilis at various defined shear levels. We show that, contrary to expectations, shear inhibits sporulation. We found an inverse correlation between the shear rate-dependent specific intracellular reactive oxygen species level (siROS), and the sporulation extent. A 10-fold increase in siROS resulted in about 17-fold decrease in sporulation extent. The involvement of reactive oxygen species (ROS) in sporulation was unknown thus far. Further, through experiments that specifically increased and reduced intracellular ROS (iROS), we established that siROS is responsible for the inhibition of sporulation under shear stress. In addition, we found that shear induced siROS regulated the expression levels of the general stress proteins Ctc and sigma(B). Based on the above, we hypothesize that siROS may regulate suppression of sporulation under high shear by altering sigma(B) and Ctc expression levels, and a model for the same is presented.

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Structure and Function of the Stressosome Signalling Hub

Pané‐Farré

Quin

Lewis

et al. 2017

Subcellular Biochemistry

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The stressosome is a multi-protein signal integration and transduction hub found in a wide range of bacterial species. The role that the stressosome plays in regulating the transcription of genes involved in the general stress response has been studied most extensively in the Gram-positive model organism Bacillus subtilis. The stressosome receives and relays the signal(s) that initiate a complex phosphorylation-dependent partner switching cascade, resulting in the activation of the alternative sigma factor σ. This sigma factor controls transcription of more than 150 genes involved in the general stress response. X-ray crystal structures of individual components of the stressosome and single-particle cryo-EM reconstructions of stressosome complexes, coupled with biochemical and single cell analyses, have permitted a detailed understanding of the dynamic signalling behaviour that arises from this multi-protein complex. Furthermore, bioinformatics analyses indicate that genetic modules encoding key stressosome proteins are found in a wide range of bacterial species, indicating an evolutionary advantage afforded by stressosome complexes. Interestingly, the genetic modules are associated with a variety of signalling modules encoding secondary messenger regulation systems, as well as classical two-component signal transduction systems, suggesting a diversification in function. In this chapter we review the current research into stressosome systems and discuss the functional implications of the unique structure of these signalling complexes.

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Effects on growth and sporulation of inactivation of a Bacillus subtilis gene (ctc) transcribed in vitro by minor vegetative cell RNA polymerases (E-σ37, E-σ32)

Cited by 23 publications

References 21 publications

The sigma B-dependent promoter of the Bacillus subtilis sigB operon is induced by heat shock

The sigma B-dependent promoter of the Bacillus subtilis sigB operon is induced by heat shock

Intracellular reactive oxygen species mediate suppression of sporulation in Bacillus subtilis under shear stress

Structure and Function of the Stressosome Signalling Hub

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