Translesion activity of PrimPol on DNA with cisplatin and DNA–protein cross-links
Abstract Human PrimPol belongs to the archaeo-eukaryotic primase superfamily of primases and is involved in de novo DNA synthesis downstream of blocking DNA lesions and non-B DNA structures. PrimPol possesses both DNA/RNA primase and DNA polymerase activities, and also bypasses a number of DNA lesio...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2021-09-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-021-96692-y |
| _version_ | 1818462155121360896 |
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| author | Elizaveta O. Boldinova Anna V. Yudkina Evgeniy S. Shilkin Diana I. Gagarinskaya Andrey G. Baranovskiy Tahir H. Tahirov Dmitry O. Zharkov Alena V. Makarova |
| author_facet | Elizaveta O. Boldinova Anna V. Yudkina Evgeniy S. Shilkin Diana I. Gagarinskaya Andrey G. Baranovskiy Tahir H. Tahirov Dmitry O. Zharkov Alena V. Makarova |
| author_sort | Elizaveta O. Boldinova |
| collection | DOAJ |
| description | Abstract Human PrimPol belongs to the archaeo-eukaryotic primase superfamily of primases and is involved in de novo DNA synthesis downstream of blocking DNA lesions and non-B DNA structures. PrimPol possesses both DNA/RNA primase and DNA polymerase activities, and also bypasses a number of DNA lesions in vitro. In this work, we have analyzed translesion synthesis activity of PrimPol in vitro on DNA with an 1,2-intrastrand cisplatin cross-link (1,2-GG CisPt CL) or a model DNA–protein cross-link (DpCL). PrimPol was capable of the 1,2-GG CisPt CL bypass in the presence of Mn2+ ions and preferentially incorporated two complementary dCMPs opposite the lesion. Nucleotide incorporation was stimulated by PolDIP2, and yeast Pol ζ efficiently extended from the nucleotides inserted opposite the 1,2-GG CisPt CL in vitro. DpCLs significantly blocked the DNA polymerase activity and strand displacement synthesis of PrimPol. However, PrimPol was able to reach the DpCL site in single strand template DNA in the presence of both Mg2+ and Mn2+ ions despite the presence of the bulky protein obstacle. |
| first_indexed | 2024-12-14T23:57:32Z |
| format | Article |
| id | doaj.art-4385708681084e81aa35235773846c76 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-12-14T23:57:32Z |
| publishDate | 2021-09-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-4385708681084e81aa35235773846c762022-12-21T22:43:03ZengNature PortfolioScientific Reports2045-23222021-09-0111111210.1038/s41598-021-96692-yTranslesion activity of PrimPol on DNA with cisplatin and DNA–protein cross-linksElizaveta O. Boldinova0Anna V. Yudkina1Evgeniy S. Shilkin2Diana I. Gagarinskaya3Andrey G. Baranovskiy4Tahir H. Tahirov5Dmitry O. Zharkov6Alena V. Makarova7Institute of Molecular Genetics, National Research Center «Kurchatov Institute»Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of SciencesInstitute of Molecular Genetics, National Research Center «Kurchatov Institute»Institute of Molecular Genetics, National Research Center «Kurchatov Institute»Eppley Institute for Research in Cancer and Allied Diseases, Fred & Pamela Buffett Cancer Center, University of Nebraska Medical CenterEppley Institute for Research in Cancer and Allied Diseases, Fred & Pamela Buffett Cancer Center, University of Nebraska Medical CenterInstitute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of SciencesInstitute of Molecular Genetics, National Research Center «Kurchatov Institute»Abstract Human PrimPol belongs to the archaeo-eukaryotic primase superfamily of primases and is involved in de novo DNA synthesis downstream of blocking DNA lesions and non-B DNA structures. PrimPol possesses both DNA/RNA primase and DNA polymerase activities, and also bypasses a number of DNA lesions in vitro. In this work, we have analyzed translesion synthesis activity of PrimPol in vitro on DNA with an 1,2-intrastrand cisplatin cross-link (1,2-GG CisPt CL) or a model DNA–protein cross-link (DpCL). PrimPol was capable of the 1,2-GG CisPt CL bypass in the presence of Mn2+ ions and preferentially incorporated two complementary dCMPs opposite the lesion. Nucleotide incorporation was stimulated by PolDIP2, and yeast Pol ζ efficiently extended from the nucleotides inserted opposite the 1,2-GG CisPt CL in vitro. DpCLs significantly blocked the DNA polymerase activity and strand displacement synthesis of PrimPol. However, PrimPol was able to reach the DpCL site in single strand template DNA in the presence of both Mg2+ and Mn2+ ions despite the presence of the bulky protein obstacle.https://doi.org/10.1038/s41598-021-96692-y |
| spellingShingle | Elizaveta O. Boldinova Anna V. Yudkina Evgeniy S. Shilkin Diana I. Gagarinskaya Andrey G. Baranovskiy Tahir H. Tahirov Dmitry O. Zharkov Alena V. Makarova Translesion activity of PrimPol on DNA with cisplatin and DNA–protein cross-links Scientific Reports |
| title | Translesion activity of PrimPol on DNA with cisplatin and DNA–protein cross-links |
| title_full | Translesion activity of PrimPol on DNA with cisplatin and DNA–protein cross-links |
| title_fullStr | Translesion activity of PrimPol on DNA with cisplatin and DNA–protein cross-links |
| title_full_unstemmed | Translesion activity of PrimPol on DNA with cisplatin and DNA–protein cross-links |
| title_short | Translesion activity of PrimPol on DNA with cisplatin and DNA–protein cross-links |
| title_sort | translesion activity of primpol on dna with cisplatin and dna protein cross links |
| url | https://doi.org/10.1038/s41598-021-96692-y |
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