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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...

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Autores principales: Wei, Wenping, Jiang, Xiaofang, Zhang, Li, Yan, Yunjun, Yan, Jinyong, Xu, Li, Gao, Chun-Hui, Yang, Min
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
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.
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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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