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

A. K. Benson, W. G. Haldenwang

Research output: Contribution to journalArticle

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Abstract

σ(B), 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 σ(B) 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 σ(B) but not RsbV, a positive regulator normally essential for σ(B)- 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 σ(B)-dependent promoter. The level of mRNA initiating at the σ(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 major 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 σ(B) 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.

Original languageEnglish (US)
Pages (from-to)1929-1935
Number of pages7
JournalJournal of bacteriology
Volume175
Issue number7
StatePublished - Jan 1 1993

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Operon
Bacillus subtilis
Shock
Hot Temperature
Proteins
Western Blotting
Sigma Factor
Monoclonal Antibodies
RNA
Messenger RNA
Temperature
Genes

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

Cite this

The σ(B)-dependent promoter of the Bacillus subtilis sigB operon is induced by heat shock. / Benson, A. K.; Haldenwang, W. G.

In: Journal of bacteriology, Vol. 175, No. 7, 01.01.1993, p. 1929-1935.

Research output: Contribution to journalArticle

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N2 - σ(B), 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 σ(B) 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 σ(B) but not RsbV, a positive regulator normally essential for σ(B)- 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 σ(B)-dependent promoter. The level of mRNA initiating at the σ(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 major 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 σ(B) 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.

AB - σ(B), 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 σ(B) 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 σ(B) but not RsbV, a positive regulator normally essential for σ(B)- 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 σ(B)-dependent promoter. The level of mRNA initiating at the σ(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 major 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 σ(B) 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.

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