RNase HI Is Essential for Survival of Mycobacterium smegmatis
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Data
2015-05-12Autor
Minias, Alina E.
Brzostek, Anna M.
Korycka- Machala, Malgorzata
Dziadek, Bozena
Minias, Piotr
Rajagopalan, Malini
Madiraju, Murty
Dziadek, Jaroslaw
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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.
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