Mutations conferring amino acid residue substitutions in the carboxy-terminal domain of RNA polymerase α can suppress clpX and clpP with respect to developmentally regulated transcription in Bacillus subtilis

Michiko Nakano, Yi Zhu, Jiajian Liu, Dindo Y. Reyes, Hirofumi Yoshikawa, Peter Zuber

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Abstract

The Bacillus subtilis clpX and clpP genes are the sites of pleiotropic mutations that adversely affect growth on a variety of media and impair developmental processes such as sporulation and competence development. ClpX is necessary for the post-exponential induction of genes that require the σ(H) form of RNA polymerase for transcription. Both ClpX and ClpP are required for the activation of σ(A)-dependent transcription of the srf operon that encodes surfactin synthetase and the regulatory peptide ComS, required for the development of genetic competence. Transcription of srf is activated by the two-component regulatory system ComPA in response to the peptide pheromone, ComX, which mediates cell density-dependent control. A clpX mutant, although able to produce ComX, is unable to respond to the pheromone. A mutant allele of comP, encoding a product whose activity is independent of ComX, is not able to suppress clpX with respect to srf expression, suggesting that ClpXP acts at the level of ComA-dependent activation of srf transcription initiation. Suppressor mutations of clpX (cxs-1 and cxs-2) were isolated in screens for pseudorevertants exhibiting high levels of srf expression and σ(H)-dependent transcription respectively. One mutation, cxs-1, suppressed a clpP null mutation with respect to srf transcription, but did not overcome the block conferred by clpP on competence development and sporulation. Both cxs-1 and cxs-2 mutations map to the region of the rpoA gene encoding the RNA polymerase α C-terminal domain (αCTD). The reconstruction of the cxs-1 and cxs-2 alleles of rpoA confirmed that these mutations confer the suppressor phenotype. These findings provide further support for the hypothesis that ClpX and ClpP might be intimately associated with transcription initiation in B. subtilis.

Original languageEnglish (US)
Pages (from-to)869-884
Number of pages16
JournalMolecular Microbiology
Volume37
Issue number4
DOIs
StatePublished - 2000

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Amino Acid Substitution
DNA-Directed RNA Polymerases
Bacillus subtilis
Mental Competency
Genetic Suppression
Mutation
Pheromones
Genetic Pleiotropy
Alleles
RNA Polymerase III
Peptides
Operon
Coma
Transcriptional Activation
Genes
Cell Count
Phenotype
Growth

ASJC Scopus subject areas

  • Molecular Biology
  • Microbiology

Cite this

@article{37050ee641074a20943bb6fbc522f14c,
title = "Mutations conferring amino acid residue substitutions in the carboxy-terminal domain of RNA polymerase α can suppress clpX and clpP with respect to developmentally regulated transcription in Bacillus subtilis",
abstract = "The Bacillus subtilis clpX and clpP genes are the sites of pleiotropic mutations that adversely affect growth on a variety of media and impair developmental processes such as sporulation and competence development. ClpX is necessary for the post-exponential induction of genes that require the σ(H) form of RNA polymerase for transcription. Both ClpX and ClpP are required for the activation of σ(A)-dependent transcription of the srf operon that encodes surfactin synthetase and the regulatory peptide ComS, required for the development of genetic competence. Transcription of srf is activated by the two-component regulatory system ComPA in response to the peptide pheromone, ComX, which mediates cell density-dependent control. A clpX mutant, although able to produce ComX, is unable to respond to the pheromone. A mutant allele of comP, encoding a product whose activity is independent of ComX, is not able to suppress clpX with respect to srf expression, suggesting that ClpXP acts at the level of ComA-dependent activation of srf transcription initiation. Suppressor mutations of clpX (cxs-1 and cxs-2) were isolated in screens for pseudorevertants exhibiting high levels of srf expression and σ(H)-dependent transcription respectively. One mutation, cxs-1, suppressed a clpP null mutation with respect to srf transcription, but did not overcome the block conferred by clpP on competence development and sporulation. Both cxs-1 and cxs-2 mutations map to the region of the rpoA gene encoding the RNA polymerase α C-terminal domain (αCTD). The reconstruction of the cxs-1 and cxs-2 alleles of rpoA confirmed that these mutations confer the suppressor phenotype. These findings provide further support for the hypothesis that ClpX and ClpP might be intimately associated with transcription initiation in B. subtilis.",
author = "Michiko Nakano and Yi Zhu and Jiajian Liu and Reyes, {Dindo Y.} and Hirofumi Yoshikawa and Peter Zuber",
year = "2000",
doi = "10.1046/j.1365-2958.2000.02052.x",
language = "English (US)",
volume = "37",
pages = "869--884",
journal = "Molecular Microbiology",
issn = "0950-382X",
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TY - JOUR

T1 - Mutations conferring amino acid residue substitutions in the carboxy-terminal domain of RNA polymerase α can suppress clpX and clpP with respect to developmentally regulated transcription in Bacillus subtilis

AU - Nakano, Michiko

AU - Zhu, Yi

AU - Liu, Jiajian

AU - Reyes, Dindo Y.

AU - Yoshikawa, Hirofumi

AU - Zuber, Peter

PY - 2000

Y1 - 2000

N2 - The Bacillus subtilis clpX and clpP genes are the sites of pleiotropic mutations that adversely affect growth on a variety of media and impair developmental processes such as sporulation and competence development. ClpX is necessary for the post-exponential induction of genes that require the σ(H) form of RNA polymerase for transcription. Both ClpX and ClpP are required for the activation of σ(A)-dependent transcription of the srf operon that encodes surfactin synthetase and the regulatory peptide ComS, required for the development of genetic competence. Transcription of srf is activated by the two-component regulatory system ComPA in response to the peptide pheromone, ComX, which mediates cell density-dependent control. A clpX mutant, although able to produce ComX, is unable to respond to the pheromone. A mutant allele of comP, encoding a product whose activity is independent of ComX, is not able to suppress clpX with respect to srf expression, suggesting that ClpXP acts at the level of ComA-dependent activation of srf transcription initiation. Suppressor mutations of clpX (cxs-1 and cxs-2) were isolated in screens for pseudorevertants exhibiting high levels of srf expression and σ(H)-dependent transcription respectively. One mutation, cxs-1, suppressed a clpP null mutation with respect to srf transcription, but did not overcome the block conferred by clpP on competence development and sporulation. Both cxs-1 and cxs-2 mutations map to the region of the rpoA gene encoding the RNA polymerase α C-terminal domain (αCTD). The reconstruction of the cxs-1 and cxs-2 alleles of rpoA confirmed that these mutations confer the suppressor phenotype. These findings provide further support for the hypothesis that ClpX and ClpP might be intimately associated with transcription initiation in B. subtilis.

AB - The Bacillus subtilis clpX and clpP genes are the sites of pleiotropic mutations that adversely affect growth on a variety of media and impair developmental processes such as sporulation and competence development. ClpX is necessary for the post-exponential induction of genes that require the σ(H) form of RNA polymerase for transcription. Both ClpX and ClpP are required for the activation of σ(A)-dependent transcription of the srf operon that encodes surfactin synthetase and the regulatory peptide ComS, required for the development of genetic competence. Transcription of srf is activated by the two-component regulatory system ComPA in response to the peptide pheromone, ComX, which mediates cell density-dependent control. A clpX mutant, although able to produce ComX, is unable to respond to the pheromone. A mutant allele of comP, encoding a product whose activity is independent of ComX, is not able to suppress clpX with respect to srf expression, suggesting that ClpXP acts at the level of ComA-dependent activation of srf transcription initiation. Suppressor mutations of clpX (cxs-1 and cxs-2) were isolated in screens for pseudorevertants exhibiting high levels of srf expression and σ(H)-dependent transcription respectively. One mutation, cxs-1, suppressed a clpP null mutation with respect to srf transcription, but did not overcome the block conferred by clpP on competence development and sporulation. Both cxs-1 and cxs-2 mutations map to the region of the rpoA gene encoding the RNA polymerase α C-terminal domain (αCTD). The reconstruction of the cxs-1 and cxs-2 alleles of rpoA confirmed that these mutations confer the suppressor phenotype. These findings provide further support for the hypothesis that ClpX and ClpP might be intimately associated with transcription initiation in B. subtilis.

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U2 - 10.1046/j.1365-2958.2000.02052.x

DO - 10.1046/j.1365-2958.2000.02052.x

M3 - Article

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EP - 884

JO - Molecular Microbiology

JF - Molecular Microbiology

SN - 0950-382X

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