Bacillus subtilis PcrA Helicase Removes Trafficking Barriers
Department of Microbial Biotechnology, Centro Nacional de Biotecnología, CNB-CSIC, 28049 Madrid, Spain
*
Author to whom correspondence should be addressed.
†
These authors contributed equally to this work.
Academic Editor: Luiz Otavio Penalva
Cells 2021, 10(4), 935; https://doi.org/10.3390/cells10040935
Received: 15 March 2021
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Revised: 12 April 2021
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Accepted: 14 April 2021
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Published: 17 April 2021
Bacillus subtilis PcrA interacts with the RNA polymerase and might contribute to mitigate replication–transcription conflicts (RTCs). We show that PcrA depletion lethality is partially suppressed by rnhB inactivation, but cell viability is significantly reduced by rnhC or dinG inactivation. Following PcrA depletion, cells lacking RnhC or DinG are extremely sensitive to DNA damage. Chromosome segregation is not further impaired by rnhB or dinG inactivation but is blocked by rnhC or recA inactivation upon PcrA depletion. Despite our efforts, we could not construct a ΔrnhC ΔrecA strain. These observations support the idea that PcrA dismantles RTCs. Purified PcrA, which binds single-stranded (ss) DNA over RNA, is a ssDNA-dependent ATPase and preferentially unwinds DNA in a 3′→5′direction. PcrA unwinds a 3′-tailed RNA of an RNA-DNA hybrid significantly faster than that of a DNA substrate. Our results suggest that a replicative stress, caused by mis-incorporated rNMPs, indirectly increases cell viability upon PcrA depletion. We propose that PcrA, in concert with RnhC or DinG, contributes to removing spontaneous or enzyme-driven R-loops, to counteract deleterious trafficking conflicts and preserve to genomic integrity.
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Keywords:
replication fork stalling; RNA polymerase backtracking; replication–transcription conflict; R-loops
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MDPI and ACS Style
Moreno-del Álamo, M.; Carrasco, B.; Torres, R.; Alonso, J.C. Bacillus subtilis PcrA Helicase Removes Trafficking Barriers. Cells 2021, 10, 935. https://doi.org/10.3390/cells10040935
AMA Style
Moreno-del Álamo M, Carrasco B, Torres R, Alonso JC. Bacillus subtilis PcrA Helicase Removes Trafficking Barriers. Cells. 2021; 10(4):935. https://doi.org/10.3390/cells10040935
Chicago/Turabian StyleMoreno-del Álamo, María, Begoña Carrasco, Rubén Torres, and Juan C. Alonso. 2021. "Bacillus subtilis PcrA Helicase Removes Trafficking Barriers" Cells 10, no. 4: 935. https://doi.org/10.3390/cells10040935
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