R-loop induced stress response by second (p)ppGpp synthetase in Mycobacterium smegmatis: Functional and domain interdependence.

Research output: Contribution to journalArticlepeer-review

Standard

Harvard

APA

Vancouver

Author

Bibtex

@article{35080d8348c7431291e30b9d6811cc22,
title = "R-loop induced stress response by second (p)ppGpp synthetase in Mycobacterium smegmatis: Functional and domain interdependence.",
abstract = "Persistent R-loops lead to replicative stress due to RNA polymerase stalling and DNA damage. RNase H enzymes facilitate the organisms to survive in the hostile condition by removing these R-loops. MS_RHII-RSD was previously identified to be the second (p)ppGpp synthetase in Mycobacterium smegmatis. The unique presence of an additional RNase HII domain raises an important question regarding the significance of this bifunctional protein. In this report, we demonstrate its ability to hydrolyze R-loops in E. coli exposed to UV stress. MS_RHII-RSD gene expression was upregulated under UV stress and this gene deleted strain showed increased R-loop accumulation as compared to the wild type. The domains in isolation are known to be inactive and the full length protein is required for its function. Domain inter-dependence studies using active site mutants reveal the necessity of a hexamer form with high alpha helical content. In previous studies, bacterial RNase type HI has been mainly implicated in R-loop hydrolysis, but in this study the RNase HII domain containing protein showed the activity. The prospective of this differential RNase HII activity is discussed. This is the first report to implicate a (p)ppGpp synthetase protein in R-loop induced stress response. This article is protected by copyright. All rights reserved.",
keywords = "Replication stress, RNase H, bifunctional protein, RNA polymerase, R-loop",
author = "Apoorva Bhatt",
year = "2016",
month = jun,
day = "28",
doi = "10.1111/mmi.13453",
language = "English",
journal = "Molecular Microbiology",
issn = "0950-382X",
publisher = "Wiley",

}

RIS

TY - JOUR

T1 - R-loop induced stress response by second (p)ppGpp synthetase in Mycobacterium smegmatis: Functional and domain interdependence.

AU - Bhatt, Apoorva

PY - 2016/6/28

Y1 - 2016/6/28

N2 - Persistent R-loops lead to replicative stress due to RNA polymerase stalling and DNA damage. RNase H enzymes facilitate the organisms to survive in the hostile condition by removing these R-loops. MS_RHII-RSD was previously identified to be the second (p)ppGpp synthetase in Mycobacterium smegmatis. The unique presence of an additional RNase HII domain raises an important question regarding the significance of this bifunctional protein. In this report, we demonstrate its ability to hydrolyze R-loops in E. coli exposed to UV stress. MS_RHII-RSD gene expression was upregulated under UV stress and this gene deleted strain showed increased R-loop accumulation as compared to the wild type. The domains in isolation are known to be inactive and the full length protein is required for its function. Domain inter-dependence studies using active site mutants reveal the necessity of a hexamer form with high alpha helical content. In previous studies, bacterial RNase type HI has been mainly implicated in R-loop hydrolysis, but in this study the RNase HII domain containing protein showed the activity. The prospective of this differential RNase HII activity is discussed. This is the first report to implicate a (p)ppGpp synthetase protein in R-loop induced stress response. This article is protected by copyright. All rights reserved.

AB - Persistent R-loops lead to replicative stress due to RNA polymerase stalling and DNA damage. RNase H enzymes facilitate the organisms to survive in the hostile condition by removing these R-loops. MS_RHII-RSD was previously identified to be the second (p)ppGpp synthetase in Mycobacterium smegmatis. The unique presence of an additional RNase HII domain raises an important question regarding the significance of this bifunctional protein. In this report, we demonstrate its ability to hydrolyze R-loops in E. coli exposed to UV stress. MS_RHII-RSD gene expression was upregulated under UV stress and this gene deleted strain showed increased R-loop accumulation as compared to the wild type. The domains in isolation are known to be inactive and the full length protein is required for its function. Domain inter-dependence studies using active site mutants reveal the necessity of a hexamer form with high alpha helical content. In previous studies, bacterial RNase type HI has been mainly implicated in R-loop hydrolysis, but in this study the RNase HII domain containing protein showed the activity. The prospective of this differential RNase HII activity is discussed. This is the first report to implicate a (p)ppGpp synthetase protein in R-loop induced stress response. This article is protected by copyright. All rights reserved.

KW - Replication stress

KW - RNase H

KW - bifunctional protein

KW - RNA polymerase

KW - R-loop

U2 - 10.1111/mmi.13453

DO - 10.1111/mmi.13453

M3 - Article

JO - Molecular Microbiology

JF - Molecular Microbiology

SN - 0950-382X

ER -