Cargando…
Conditional down-regulation of GreA impacts expression of rRNA and transcription factors, affecting Mycobacterium smegmatis survival
Gre, one of the conserved transcription factors in bacteria, modulates RNA polymerase (RNAP) activity to ensure processivity and fidelity of RNA synthesis. Gre factors regulate transcription by inducing the intrinsic-endonucleolytic activity of RNAP, allowing the enzyme to resume transcription from...
Autores principales: | , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2020
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7118132/ https://www.ncbi.nlm.nih.gov/pubmed/32242064 http://dx.doi.org/10.1038/s41598-020-62703-7 |
_version_ | 1783514496975765504 |
---|---|
author | Jha, Rajiv Kumar Udupa, Shubha Rai, Ashutosh Kumar Rani, Phoolwanti Singh, Prakruti R. Govind, Shamitha Nagaraja, Valakunja |
author_facet | Jha, Rajiv Kumar Udupa, Shubha Rai, Ashutosh Kumar Rani, Phoolwanti Singh, Prakruti R. Govind, Shamitha Nagaraja, Valakunja |
author_sort | Jha, Rajiv Kumar |
collection | PubMed |
description | Gre, one of the conserved transcription factors in bacteria, modulates RNA polymerase (RNAP) activity to ensure processivity and fidelity of RNA synthesis. Gre factors regulate transcription by inducing the intrinsic-endonucleolytic activity of RNAP, allowing the enzyme to resume transcription from the paused and arrested sites. While Escherichia coli and a number of eubacteria harbor GreA and GreB, genus mycobacteria has a single Gre (GreA). To address the importance of the GreA in growth, physiology and gene expression of Mycobacterium smegmatis, we have constructed a conditional knock-down strain of GreA. The GreA depleted strain exhibited slow growth, drastic changes in cell surface phenotype, cell death, and increased susceptibility to front-line anti-tubercular drugs. Transcripts and 2D-gel electrophoresis (2D-PAGE) analysis of the GreA conditional knock-down strain showed altered expression of the genes involved in transcription regulation. Among the genes analysed, expression of RNAP subunits (β, β’ and ω), carD, hupB, lsr2, and nusA were affected to a large extent. Severe reduction in the expression of genes of rRNA operon in the knock-down strain reveal a role for GreA in regulating the core components of the translation process. |
format | Online Article Text |
id | pubmed-7118132 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-71181322020-04-08 Conditional down-regulation of GreA impacts expression of rRNA and transcription factors, affecting Mycobacterium smegmatis survival Jha, Rajiv Kumar Udupa, Shubha Rai, Ashutosh Kumar Rani, Phoolwanti Singh, Prakruti R. Govind, Shamitha Nagaraja, Valakunja Sci Rep Article Gre, one of the conserved transcription factors in bacteria, modulates RNA polymerase (RNAP) activity to ensure processivity and fidelity of RNA synthesis. Gre factors regulate transcription by inducing the intrinsic-endonucleolytic activity of RNAP, allowing the enzyme to resume transcription from the paused and arrested sites. While Escherichia coli and a number of eubacteria harbor GreA and GreB, genus mycobacteria has a single Gre (GreA). To address the importance of the GreA in growth, physiology and gene expression of Mycobacterium smegmatis, we have constructed a conditional knock-down strain of GreA. The GreA depleted strain exhibited slow growth, drastic changes in cell surface phenotype, cell death, and increased susceptibility to front-line anti-tubercular drugs. Transcripts and 2D-gel electrophoresis (2D-PAGE) analysis of the GreA conditional knock-down strain showed altered expression of the genes involved in transcription regulation. Among the genes analysed, expression of RNAP subunits (β, β’ and ω), carD, hupB, lsr2, and nusA were affected to a large extent. Severe reduction in the expression of genes of rRNA operon in the knock-down strain reveal a role for GreA in regulating the core components of the translation process. Nature Publishing Group UK 2020-04-02 /pmc/articles/PMC7118132/ /pubmed/32242064 http://dx.doi.org/10.1038/s41598-020-62703-7 Text en © The Author(s) 2020 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Article Jha, Rajiv Kumar Udupa, Shubha Rai, Ashutosh Kumar Rani, Phoolwanti Singh, Prakruti R. Govind, Shamitha Nagaraja, Valakunja Conditional down-regulation of GreA impacts expression of rRNA and transcription factors, affecting Mycobacterium smegmatis survival |
title | Conditional down-regulation of GreA impacts expression of rRNA and transcription factors, affecting Mycobacterium smegmatis survival |
title_full | Conditional down-regulation of GreA impacts expression of rRNA and transcription factors, affecting Mycobacterium smegmatis survival |
title_fullStr | Conditional down-regulation of GreA impacts expression of rRNA and transcription factors, affecting Mycobacterium smegmatis survival |
title_full_unstemmed | Conditional down-regulation of GreA impacts expression of rRNA and transcription factors, affecting Mycobacterium smegmatis survival |
title_short | Conditional down-regulation of GreA impacts expression of rRNA and transcription factors, affecting Mycobacterium smegmatis survival |
title_sort | conditional down-regulation of grea impacts expression of rrna and transcription factors, affecting mycobacterium smegmatis survival |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7118132/ https://www.ncbi.nlm.nih.gov/pubmed/32242064 http://dx.doi.org/10.1038/s41598-020-62703-7 |
work_keys_str_mv | AT jharajivkumar conditionaldownregulationofgreaimpactsexpressionofrrnaandtranscriptionfactorsaffectingmycobacteriumsmegmatissurvival AT udupashubha conditionaldownregulationofgreaimpactsexpressionofrrnaandtranscriptionfactorsaffectingmycobacteriumsmegmatissurvival AT raiashutoshkumar conditionaldownregulationofgreaimpactsexpressionofrrnaandtranscriptionfactorsaffectingmycobacteriumsmegmatissurvival AT raniphoolwanti conditionaldownregulationofgreaimpactsexpressionofrrnaandtranscriptionfactorsaffectingmycobacteriumsmegmatissurvival AT singhprakrutir conditionaldownregulationofgreaimpactsexpressionofrrnaandtranscriptionfactorsaffectingmycobacteriumsmegmatissurvival AT govindshamitha conditionaldownregulationofgreaimpactsexpressionofrrnaandtranscriptionfactorsaffectingmycobacteriumsmegmatissurvival AT nagarajavalakunja conditionaldownregulationofgreaimpactsexpressionofrrnaandtranscriptionfactorsaffectingmycobacteriumsmegmatissurvival |