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Regulation of CRISPR-Associated Genes by Rv1776c (CasR) in Mycobacterium tuberculosis
The CRISPR-Cas system is an adaptive immune system for many bacteria and archaea to defend against foreign nucleic acid invasion, and this system is conserved in the genome of M. tuberculosis (Mtb). Although the CRISPR-Cas system-mediated immune defense mechanism has been revealed in Mtb, the regula...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9953421/ https://www.ncbi.nlm.nih.gov/pubmed/36830769 http://dx.doi.org/10.3390/biom13020400 |
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author | Wei, Wenping Jiang, Xiaofang Zhang, Li Yan, Yunjun Yan, Jinyong Xu, Li Gao, Chun-Hui Yang, Min |
author_facet | Wei, Wenping Jiang, Xiaofang Zhang, Li Yan, Yunjun Yan, Jinyong Xu, Li Gao, Chun-Hui Yang, Min |
author_sort | Wei, Wenping |
collection | PubMed |
description | The CRISPR-Cas system is an adaptive immune system for many bacteria and archaea to defend against foreign nucleic acid invasion, and this system is conserved in the genome of M. tuberculosis (Mtb). Although the CRISPR-Cas system-mediated immune defense mechanism has been revealed in Mtb, the regulation of cas gene expression is poorly understood. In this study, we identified a transcription factor, CasR (CRISPR-associated protein repressor, encoded by Rv1776c), and it could bind to the upstream DNA sequence of the CRISPR-Cas gene cluster and regulate the expression of cas genes. EMSA and ChIP assays confirmed that CasR could interact with the upstream sequence of the csm6 promoter, both in vivo and in vitro. Furthermore, DNA footprinting assay revealed that CasR recognized a 20 bp palindromic sequence motif and negatively regulated the expression of csm6. In conclusion, our research elucidates the regulatory effect of CasR on the expression of CRISPR-associated genes in mycobacteria, thus providing insight into gene expression regulation of the CRISPR-Cas system. |
format | Online Article Text |
id | pubmed-9953421 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-99534212023-02-25 Regulation of CRISPR-Associated Genes by Rv1776c (CasR) in Mycobacterium tuberculosis Wei, Wenping Jiang, Xiaofang Zhang, Li Yan, Yunjun Yan, Jinyong Xu, Li Gao, Chun-Hui Yang, Min Biomolecules Article The CRISPR-Cas system is an adaptive immune system for many bacteria and archaea to defend against foreign nucleic acid invasion, and this system is conserved in the genome of M. tuberculosis (Mtb). Although the CRISPR-Cas system-mediated immune defense mechanism has been revealed in Mtb, the regulation of cas gene expression is poorly understood. In this study, we identified a transcription factor, CasR (CRISPR-associated protein repressor, encoded by Rv1776c), and it could bind to the upstream DNA sequence of the CRISPR-Cas gene cluster and regulate the expression of cas genes. EMSA and ChIP assays confirmed that CasR could interact with the upstream sequence of the csm6 promoter, both in vivo and in vitro. Furthermore, DNA footprinting assay revealed that CasR recognized a 20 bp palindromic sequence motif and negatively regulated the expression of csm6. In conclusion, our research elucidates the regulatory effect of CasR on the expression of CRISPR-associated genes in mycobacteria, thus providing insight into gene expression regulation of the CRISPR-Cas system. MDPI 2023-02-20 /pmc/articles/PMC9953421/ /pubmed/36830769 http://dx.doi.org/10.3390/biom13020400 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Wei, Wenping Jiang, Xiaofang Zhang, Li Yan, Yunjun Yan, Jinyong Xu, Li Gao, Chun-Hui Yang, Min Regulation of CRISPR-Associated Genes by Rv1776c (CasR) in Mycobacterium tuberculosis |
title | Regulation of CRISPR-Associated Genes by Rv1776c (CasR) in Mycobacterium tuberculosis |
title_full | Regulation of CRISPR-Associated Genes by Rv1776c (CasR) in Mycobacterium tuberculosis |
title_fullStr | Regulation of CRISPR-Associated Genes by Rv1776c (CasR) in Mycobacterium tuberculosis |
title_full_unstemmed | Regulation of CRISPR-Associated Genes by Rv1776c (CasR) in Mycobacterium tuberculosis |
title_short | Regulation of CRISPR-Associated Genes by Rv1776c (CasR) in Mycobacterium tuberculosis |
title_sort | regulation of crispr-associated genes by rv1776c (casr) in mycobacterium tuberculosis |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9953421/ https://www.ncbi.nlm.nih.gov/pubmed/36830769 http://dx.doi.org/10.3390/biom13020400 |
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