Transcriptional Regulation and Mechanism of SigN (ZpdN), a pBS32-Encoded Sigma Factor in <named-content content-type="genus-species">Bacillus subtilis</named-content>

ABSTRACT Laboratory strains of Bacillus subtilis encode many alternative sigma factors, each dedicated to expressing a unique regulon such as those involved in stress resistance, sporulation, and motility. The ancestral strain of B. subtilis also encodes an additional sigma factor homolog, ZpdN, not...

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Autores principales:	Aisha T. Burton, Aaron DeLoughery, Gene-Wei Li, Daniel B. Kearns
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2019
Materias:	sigma factor RNAP plasmid LexA cell death RNA polymerases Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/8ead7ef4dd394801905a39f44cd41b1a
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spelling	oai:doaj.org-article:8ead7ef4dd394801905a39f44cd41b1a2021-11-15T15:59:42ZTranscriptional Regulation and Mechanism of SigN (ZpdN), a pBS32-Encoded Sigma Factor in <named-content content-type="genus-species">Bacillus subtilis</named-content>10.1128/mBio.01899-192150-7511https://doaj.org/article/8ead7ef4dd394801905a39f44cd41b1a2019-10-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.01899-19https://doaj.org/toc/2150-7511ABSTRACT Laboratory strains of Bacillus subtilis encode many alternative sigma factors, each dedicated to expressing a unique regulon such as those involved in stress resistance, sporulation, and motility. The ancestral strain of B. subtilis also encodes an additional sigma factor homolog, ZpdN, not found in lab strains due to being encoded on the large, low-copy-number plasmid pBS32, which was lost during domestication. DNA damage triggers pBS32 hyperreplication and cell death in a manner that depends on ZpdN, but how ZpdN mediates these effects is unknown. Here, we show that ZpdN is a bona fide sigma factor that can direct RNA polymerase to transcribe ZpdN-dependent genes, and we rename ZpdN SigN accordingly. Rend-seq (end-enriched transcriptome sequencing) analysis was used to determine the SigN regulon on pBS32, and the 5′ ends of transcripts were used to predict the SigN consensus sequence. Finally, we characterize the regulation of SigN itself and show that it is transcribed by at least three promoters: PsigN1, a strong SigA-dependent LexA-repressed promoter; PsigN2, a weak SigA-dependent constitutive promoter; and PsigN3, a SigN-dependent promoter. Thus, in response to DNA damage SigN is derepressed and then experiences positive feedback. How cells die in a pBS32-dependent manner remains unknown, but we predict that death is the product of expressing one or more genes in the SigN regulon. IMPORTANCE Sigma factors are utilized by bacteria to control and regulate gene expression. Some sigma factors are activated during times of stress to ensure the survival of the bacterium. Here, we report the presence of a sigma factor that is encoded on a plasmid that leads to cellular death after DNA damage.Aisha T. BurtonAaron DeLougheryGene-Wei LiDaniel B. KearnsAmerican Society for Microbiologyarticlesigma factorRNAPplasmidLexAcell deathRNA polymerasesMicrobiologyQR1-502ENmBio, Vol 10, Iss 5 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	sigma factor RNAP plasmid LexA cell death RNA polymerases Microbiology QR1-502
spellingShingle	sigma factor RNAP plasmid LexA cell death RNA polymerases Microbiology QR1-502 Aisha T. Burton Aaron DeLoughery Gene-Wei Li Daniel B. Kearns Transcriptional Regulation and Mechanism of SigN (ZpdN), a pBS32-Encoded Sigma Factor in <named-content content-type="genus-species">Bacillus subtilis</named-content>
description	ABSTRACT Laboratory strains of Bacillus subtilis encode many alternative sigma factors, each dedicated to expressing a unique regulon such as those involved in stress resistance, sporulation, and motility. The ancestral strain of B. subtilis also encodes an additional sigma factor homolog, ZpdN, not found in lab strains due to being encoded on the large, low-copy-number plasmid pBS32, which was lost during domestication. DNA damage triggers pBS32 hyperreplication and cell death in a manner that depends on ZpdN, but how ZpdN mediates these effects is unknown. Here, we show that ZpdN is a bona fide sigma factor that can direct RNA polymerase to transcribe ZpdN-dependent genes, and we rename ZpdN SigN accordingly. Rend-seq (end-enriched transcriptome sequencing) analysis was used to determine the SigN regulon on pBS32, and the 5′ ends of transcripts were used to predict the SigN consensus sequence. Finally, we characterize the regulation of SigN itself and show that it is transcribed by at least three promoters: PsigN1, a strong SigA-dependent LexA-repressed promoter; PsigN2, a weak SigA-dependent constitutive promoter; and PsigN3, a SigN-dependent promoter. Thus, in response to DNA damage SigN is derepressed and then experiences positive feedback. How cells die in a pBS32-dependent manner remains unknown, but we predict that death is the product of expressing one or more genes in the SigN regulon. IMPORTANCE Sigma factors are utilized by bacteria to control and regulate gene expression. Some sigma factors are activated during times of stress to ensure the survival of the bacterium. Here, we report the presence of a sigma factor that is encoded on a plasmid that leads to cellular death after DNA damage.
format	article
author	Aisha T. Burton Aaron DeLoughery Gene-Wei Li Daniel B. Kearns
author_facet	Aisha T. Burton Aaron DeLoughery Gene-Wei Li Daniel B. Kearns
author_sort	Aisha T. Burton
title	Transcriptional Regulation and Mechanism of SigN (ZpdN), a pBS32-Encoded Sigma Factor in <named-content content-type="genus-species">Bacillus subtilis</named-content>
title_short	Transcriptional Regulation and Mechanism of SigN (ZpdN), a pBS32-Encoded Sigma Factor in <named-content content-type="genus-species">Bacillus subtilis</named-content>
title_full	Transcriptional Regulation and Mechanism of SigN (ZpdN), a pBS32-Encoded Sigma Factor in <named-content content-type="genus-species">Bacillus subtilis</named-content>
title_fullStr	Transcriptional Regulation and Mechanism of SigN (ZpdN), a pBS32-Encoded Sigma Factor in <named-content content-type="genus-species">Bacillus subtilis</named-content>
title_full_unstemmed	Transcriptional Regulation and Mechanism of SigN (ZpdN), a pBS32-Encoded Sigma Factor in <named-content content-type="genus-species">Bacillus subtilis</named-content>
title_sort	transcriptional regulation and mechanism of sign (zpdn), a pbs32-encoded sigma factor in <named-content content-type="genus-species">bacillus subtilis</named-content>
publisher	American Society for Microbiology
publishDate	2019
url	https://doaj.org/article/8ead7ef4dd394801905a39f44cd41b1a
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Transcriptional Regulation and Mechanism of SigN (ZpdN), a pBS32-Encoded Sigma Factor in <named-content content-type="genus-species">Bacillus subtilis</named-content>

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