Cargando…
Transcriptional Regulation and Mechanism of SigN (ZpdN), a pBS32-Encoded Sigma Factor in Bacillus subtilis
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...
Autores principales: | , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society for Microbiology
2019
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6751061/ https://www.ncbi.nlm.nih.gov/pubmed/31530675 http://dx.doi.org/10.1128/mBio.01899-19 |
_version_ | 1783452549149360128 |
---|---|
author | Burton, Aisha T. DeLoughery, Aaron Li, Gene-Wei Kearns, Daniel B. |
author_facet | Burton, Aisha T. DeLoughery, Aaron Li, Gene-Wei Kearns, Daniel B. |
author_sort | Burton, Aisha T. |
collection | PubMed |
description | 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: P(sigN1), a strong SigA-dependent LexA-repressed promoter; P(sigN2), a weak SigA-dependent constitutive promoter; and P(sigN3), 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. |
format | Online Article Text |
id | pubmed-6751061 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | American Society for Microbiology |
record_format | MEDLINE/PubMed |
spelling | pubmed-67510612019-09-24 Transcriptional Regulation and Mechanism of SigN (ZpdN), a pBS32-Encoded Sigma Factor in Bacillus subtilis Burton, Aisha T. DeLoughery, Aaron Li, Gene-Wei Kearns, Daniel B. mBio Research Article 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: P(sigN1), a strong SigA-dependent LexA-repressed promoter; P(sigN2), a weak SigA-dependent constitutive promoter; and P(sigN3), 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. American Society for Microbiology 2019-09-17 /pmc/articles/PMC6751061/ /pubmed/31530675 http://dx.doi.org/10.1128/mBio.01899-19 Text en Copyright © 2019 Burton et al. https://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Research Article Burton, Aisha T. DeLoughery, Aaron Li, Gene-Wei Kearns, Daniel B. Transcriptional Regulation and Mechanism of SigN (ZpdN), a pBS32-Encoded Sigma Factor in Bacillus subtilis |
title | Transcriptional Regulation and Mechanism of SigN (ZpdN), a pBS32-Encoded Sigma Factor in Bacillus subtilis |
title_full | Transcriptional Regulation and Mechanism of SigN (ZpdN), a pBS32-Encoded Sigma Factor in Bacillus subtilis |
title_fullStr | Transcriptional Regulation and Mechanism of SigN (ZpdN), a pBS32-Encoded Sigma Factor in Bacillus subtilis |
title_full_unstemmed | Transcriptional Regulation and Mechanism of SigN (ZpdN), a pBS32-Encoded Sigma Factor in Bacillus subtilis |
title_short | Transcriptional Regulation and Mechanism of SigN (ZpdN), a pBS32-Encoded Sigma Factor in Bacillus subtilis |
title_sort | transcriptional regulation and mechanism of sign (zpdn), a pbs32-encoded sigma factor in bacillus subtilis |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6751061/ https://www.ncbi.nlm.nih.gov/pubmed/31530675 http://dx.doi.org/10.1128/mBio.01899-19 |
work_keys_str_mv | AT burtonaishat transcriptionalregulationandmechanismofsignzpdnapbs32encodedsigmafactorinbacillussubtilis AT delougheryaaron transcriptionalregulationandmechanismofsignzpdnapbs32encodedsigmafactorinbacillussubtilis AT ligenewei transcriptionalregulationandmechanismofsignzpdnapbs32encodedsigmafactorinbacillussubtilis AT kearnsdanielb transcriptionalregulationandmechanismofsignzpdnapbs32encodedsigmafactorinbacillussubtilis |