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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, Alexander V. Mazin
doi: https://doi.org/10.1101/2020.02.11.943977
Olga M. Mazina
1Department of Biochemistry and Molecular Biology, Drexel University College of Medicine, Philadelphia, PA 19102, USA
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Srinivas Somarowthu
1Department of Biochemistry and Molecular Biology, Drexel University College of Medicine, Philadelphia, PA 19102, USA
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Lyudmila Y. Kadyrova
2Department of Biochemistry and Molecular Biology, Southern Illinois University School of Medicine, Carbondale, IL, 62901, USA
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Andrey G. Baranovskiy
3Eppley Institute for Research in Cancer and Allied Diseases, Fred and Pamela Buffett Cancer Center, Omaha, NE, 68198, USA
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Tahir H. Tahirov
3Eppley Institute for Research in Cancer and Allied Diseases, Fred and Pamela Buffett Cancer Center, Omaha, NE, 68198, USA
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Farid A. Kadyrov
2Department of Biochemistry and Molecular Biology, Southern Illinois University School of Medicine, Carbondale, IL, 62901, USA
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Alexander V. Mazin
1Department of Biochemistry and Molecular Biology, Drexel University College of Medicine, Philadelphia, PA 19102, USA
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  • For correspondence: avm28@drexel.edu
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SUMMARY

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. Surprisingly, we found here that RPA binds RNA in vitro with high affinity. Using native RIP method, we isolated RNA-RPA complexes from human cells. Furthermore, RPA promotes R-loop formation between RNA and homologous dsDNA. R-loops, the three-stranded nucleic acid structure consisting of an RNA-DNA hybrid and the displaced ssDNA strand, are common in human genome. R-loops may play an important role in transcription-coupled homologous recombination and DNA replication restart. We reconstituted the process of replication restart in vitro using RPA-generated R-loops and human DNA polymerases. These findings indicate that RPA may play a role in RNA metabolism and suggest a mechanism of genome maintenance that depends on RPA and RNA.

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Posted February 12, 2020.
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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, Alexander V. Mazin
bioRxiv 2020.02.11.943977; doi: https://doi.org/10.1101/2020.02.11.943977
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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, Alexander V. Mazin
bioRxiv 2020.02.11.943977; doi: https://doi.org/10.1101/2020.02.11.943977

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