RNase HI Is Essential for Survival of Mycobacterium smegmatis.
RNases H are involved in the removal of RNA from RNA/DNA hybrids. Type I RNases H are thought to recognize and cleave the RNA/DNA duplex when at least four ribonucleotides are present. Here we investigated the importance of RNase H type I encoding genes for model organism Mycobacterium smegmatis. By...
Main Authors: | , , , , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
Public Library of Science (PLoS)
2015-01-01
|
Series: | PLoS ONE |
Online Access: | http://europepmc.org/articles/PMC4429107?pdf=render |
id |
doaj-bdea6419b30e4d7ab256481b0d20466b |
---|---|
record_format |
Article |
spelling |
doaj-bdea6419b30e4d7ab256481b0d20466b2020-11-24T21:24:26ZengPublic Library of Science (PLoS)PLoS ONE1932-62032015-01-01105e012626010.1371/journal.pone.0126260RNase HI Is Essential for Survival of Mycobacterium smegmatis.Alina E MiniasAnna M BrzostekMalgorzata Korycka-MachalaBozena DziadekPiotr MiniasMalini RajagopalanMurty MadirajuJaroslaw DziadekRNases H are involved in the removal of RNA from RNA/DNA hybrids. Type I RNases H are thought to recognize and cleave the RNA/DNA duplex when at least four ribonucleotides are present. Here we investigated the importance of RNase H type I encoding genes for model organism Mycobacterium smegmatis. By performing gene replacement through homologous recombination, we demonstrate that each of the two presumable RNase H type I encoding genes, rnhA and MSMEG4305, can be removed from M. smegmatis genome without affecting the growth rate of the mutant. Further, we demonstrate that deletion of both RNases H type I encoding genes in M. smegmatis leads to synthetic lethality. Finally, we question the possibility of existence of RNase HI related alternative mode of initiation of DNA replication in M. smegmatis, the process initially discovered in Escherichia coli. We suspect that synthetic lethality of double mutant lacking RNases H type I is caused by formation of R-loops leading to collapse of replication forks. We report Mycobacterium smegmatis as the first bacterial species, where function of RNase H type I has been found essential.http://europepmc.org/articles/PMC4429107?pdf=render |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Alina E Minias Anna M Brzostek Malgorzata Korycka-Machala Bozena Dziadek Piotr Minias Malini Rajagopalan Murty Madiraju Jaroslaw Dziadek |
spellingShingle |
Alina E Minias Anna M Brzostek Malgorzata Korycka-Machala Bozena Dziadek Piotr Minias Malini Rajagopalan Murty Madiraju Jaroslaw Dziadek RNase HI Is Essential for Survival of Mycobacterium smegmatis. PLoS ONE |
author_facet |
Alina E Minias Anna M Brzostek Malgorzata Korycka-Machala Bozena Dziadek Piotr Minias Malini Rajagopalan Murty Madiraju Jaroslaw Dziadek |
author_sort |
Alina E Minias |
title |
RNase HI Is Essential for Survival of Mycobacterium smegmatis. |
title_short |
RNase HI Is Essential for Survival of Mycobacterium smegmatis. |
title_full |
RNase HI Is Essential for Survival of Mycobacterium smegmatis. |
title_fullStr |
RNase HI Is Essential for Survival of Mycobacterium smegmatis. |
title_full_unstemmed |
RNase HI Is Essential for Survival of Mycobacterium smegmatis. |
title_sort |
rnase hi is essential for survival of mycobacterium smegmatis. |
publisher |
Public Library of Science (PLoS) |
series |
PLoS ONE |
issn |
1932-6203 |
publishDate |
2015-01-01 |
description |
RNases H are involved in the removal of RNA from RNA/DNA hybrids. Type I RNases H are thought to recognize and cleave the RNA/DNA duplex when at least four ribonucleotides are present. Here we investigated the importance of RNase H type I encoding genes for model organism Mycobacterium smegmatis. By performing gene replacement through homologous recombination, we demonstrate that each of the two presumable RNase H type I encoding genes, rnhA and MSMEG4305, can be removed from M. smegmatis genome without affecting the growth rate of the mutant. Further, we demonstrate that deletion of both RNases H type I encoding genes in M. smegmatis leads to synthetic lethality. Finally, we question the possibility of existence of RNase HI related alternative mode of initiation of DNA replication in M. smegmatis, the process initially discovered in Escherichia coli. We suspect that synthetic lethality of double mutant lacking RNases H type I is caused by formation of R-loops leading to collapse of replication forks. We report Mycobacterium smegmatis as the first bacterial species, where function of RNase H type I has been found essential. |
url |
http://europepmc.org/articles/PMC4429107?pdf=render |
work_keys_str_mv |
AT alinaeminias rnasehiisessentialforsurvivalofmycobacteriumsmegmatis AT annambrzostek rnasehiisessentialforsurvivalofmycobacteriumsmegmatis AT malgorzatakoryckamachala rnasehiisessentialforsurvivalofmycobacteriumsmegmatis AT bozenadziadek rnasehiisessentialforsurvivalofmycobacteriumsmegmatis AT piotrminias rnasehiisessentialforsurvivalofmycobacteriumsmegmatis AT malinirajagopalan rnasehiisessentialforsurvivalofmycobacteriumsmegmatis AT murtymadiraju rnasehiisessentialforsurvivalofmycobacteriumsmegmatis AT jaroslawdziadek rnasehiisessentialforsurvivalofmycobacteriumsmegmatis |
_version_ |
1725988206658191360 |