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...
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| Format: | Article |
| Language: | English |
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American Society for Microbiology
2020
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| Online Access: | https://hdl.handle.net/1721.1/125104 |
| _version_ | 1826205721796018176 |
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| author | DeLoughery, Aaron Li, Gene-Wei |
| author2 | Massachusetts Institute of Technology. Department of Biology |
| author_facet | Massachusetts Institute of Technology. Department of Biology DeLoughery, Aaron Li, Gene-Wei |
| author_sort | DeLoughery, Aaron |
| collection | MIT |
| 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: PsigN1, a strong SigA-dependent LexArepressed 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. |
| first_indexed | 2024-09-23T13:17:33Z |
| format | Article |
| id | mit-1721.1/125104 |
| institution | Massachusetts Institute of Technology |
| language | English |
| last_indexed | 2024-09-23T13:17:33Z |
| publishDate | 2020 |
| publisher | American Society for Microbiology |
| record_format | dspace |
| spelling | mit-1721.1/1251042022-10-01T14:20:15Z Transcriptional regulation and mechanism of sigN (ZpdN), a pBS32-encoded sigma factor in bacillus subtilis DeLoughery, Aaron Li, Gene-Wei Massachusetts Institute of Technology. Department of Biology 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 LexArepressed 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. National Institutes of Health (U.S.) (Grant 35GM124732) 2020-05-07T16:44:41Z 2020-05-07T16:44:41Z 2019-09 2020-01-23T17:45:00Z Article http://purl.org/eprint/type/JournalArticle 2150-7511 https://hdl.handle.net/1721.1/125104 Burton, Aisha T. et al. “Transcriptional regulation and mechanism of sigN (ZpdN), a pBS32-encoded sigma factor in bacillus subtilis.” MBio 10 (2019): e01899-19 © 2019 The Author(s) en 10.1128/mBio.01899-19 MBio Creative Commons Attribution 4.0 International license https://creativecommons.org/licenses/by/4.0/ application/pdf American Society for Microbiology mBio |
| spellingShingle | DeLoughery, Aaron Li, Gene-Wei 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 |
| url | https://hdl.handle.net/1721.1/125104 |
| work_keys_str_mv | AT delougheryaaron transcriptionalregulationandmechanismofsignzpdnapbs32encodedsigmafactorinbacillussubtilis AT ligenewei transcriptionalregulationandmechanismofsignzpdnapbs32encodedsigmafactorinbacillussubtilis |