Translesion activity of PrimPol on DNA with cisplatin and DNA-protein cross-links.
Translesion activity of PrimPol on DNA with cisplatin and DNA-protein cross-links.
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DOI：
10.1038/s41598-021-96692-y
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发表时间：
2021-09-02
影响因子：
4.6
通讯作者：
Makarova AV
中科院分区：
文献类型：
--
作者：
Boldinova EO;Yudkina AV;Shilkin ES;Gagarinskaya DI;Baranovskiy AG;Tahirov TH;Zharkov DO;Makarova AV
人PrimPol属于引物酶的古-真核引物酶超家族，并参与阻断DNA损伤和非B DNA结构下游的从头DNA合成。PrimPol具有DNA/RNA引发酶和DNA聚合酶活性，并且还在体外绕过许多DNA损伤。在这项工作中，我们分析了PrimPol在体外对具有1，2-链内顺铂交联（1，2-GG CisPt CL）或模型DNA-蛋白质交联（DpCL）的DNA的跨损伤合成活性。PrimPol能够在Mn 2+离子存在下进行1，2-GG CisPt CL旁路，并优先掺入病变对面的两个互补dCMP。通过PolDIP 2刺激核苷酸掺入，并且酵母PolDIP 2在体外从相对于1，2-GG CisPt CL插入的核苷酸有效地延伸。DpCL显著阻断PrimPol的DNA聚合酶活性和链置换合成。然而，PrimPol能够在Mg 2+和Mn 2+离子两者的存在下到达单链模板DNA中的DpCL位点，尽管存在庞大的蛋白质障碍。
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.
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影响因子：
14.9
作者：
Guilliam TA;Bailey LJ;Brissett NC;Doherty AJ
通讯作者：
Doherty AJ
影响因子：
16
作者：
Bianchi, Julie;Rudd, Sean G.;Jozwiakowski, Stanislaw K.;Bailey, Laura J.;Soura, Violetta;Taylor, Elaine;Stevanovic, Irena;Green, Andrew J.;Stracker, Travis H.;Lindsay, Howard D.;Doherty, Aidan J.
通讯作者：
Doherty, Aidan J.
DOI：
10.1007/978-1-61779-998-3_15
发表时间：
2012
期刊：
Methods in molecular biology (Clifton, N.J.)
影响因子：
--
作者：
Enoiu, Milica;Ho, The Vinh;Long, David T;Walter, Johannes C;Scharer, Orlando D
通讯作者：
Scharer, Orlando D
影响因子：
2.8
作者：
Kojima Y;Machida YJ
通讯作者：
Machida YJ
影响因子：
2.9
作者：
KOW, YW;WALLACE, SS
通讯作者：
WALLACE, SS