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Replication protein A binds RNA and promotes R-loop formation
Journal article   Open access   Peer reviewed

Replication protein A binds RNA and promotes R-loop formation

Olga M Mazina, Srinivas Somarowthu, Lyudmila Y Kadyrova, Andrey G Baranovskiy, Tahir H Tahirov, Farid A Kadyrov and Alexander V Mazin
The Journal of biological chemistry, v 295(41), pp 14203-14213
09 Oct 2020
DOI: https://doi.org/10.1074/jbc.RA120.013812
PMID: 32796030
Featured in Collection :   UN Sustainable Development Goals @ Drexel
url
https://doi.org/10.1074/jbc.ra120.013812View
Published, Version of Record (VoR)CC BY V4.0,  Open
url
https://doi.org/10.1074/jbc.RA120.013812View
Published, Version of Record (VoR) Open

Abstract

DNA - biosynthesis DNA - chemistry DNA Replication Humans Protein Binding R-Loop Structures Replication Protein A - chemistry Replication Protein A - metabolism RNA - chemistry RNA - metabolism
Replication protein A (RPA), a major eukaryotic ssDNA-binding protein, is essential for all metabolic processes that involve ssDNA, including DNA replication, repair, and damage signaling. To perform its functions, RPA binds ssDNA tightly. In contrast, it was presumed that RPA binds RNA weakly. However, recent data suggest that RPA may play a role in RNA metabolism. RPA stimulates RNA-templated DNA repair and associates with R-loops, the three-stranded structures consisting of an RNA-DNA hybrid and the displaced ssDNA strand. R-loops are common in the genomes of pro- and eukaryotes, including humans, and may play an important role in transcription-coupled homologous recombination and DNA replication restart. However, the mechanism of R-loop formation remains unknown. Here, we investigated the RNA-binding properties of human RPA and its possible role in R-loop formation. Using gel-retardation and RNA/DNA competition assays, we found that RPA binds RNA with an unexpectedly high affinity ( ≈ 100 pm). Furthermore, RPA, by forming a complex with RNA, can promote R-loop formation with homologous dsDNA. In reconstitution experiments, we showed that human DNA polymerases can utilize RPA-generated R-loops for initiation of DNA synthesis, mimicking the process of replication restart These results demonstrate that RPA binds RNA with high affinity, supporting the role of this protein in RNA metabolism and suggesting a mechanism of genome maintenance that depends on RPA-mediated DNA replication restart.

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34 citations in Web of Science
33 citations in Scopus

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Collaboration types
Domestic collaboration
Web of Science research areas
Biochemistry & Molecular Biology
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