Werner Helicase Control of Human Papillomavirus 16 E1-E2 DNA Replication Is Regulated by SIRT1 Deacetylation
Werner Helicase Control of Human Papillomavirus 16 E1-E2 DNA Replication Is Regulated by SIRT1 Deacetylation
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DOI：
10.1128/mbio.00263-19
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发表时间：
2019-03-01
期刊：
影响因子：
6.4
通讯作者：
Morgan, Iain M.
中科院分区：
文献类型：
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作者：
Das, Dipon;Bristol, Molly L.;Morgan, Iain M.
人乳头瘤病毒（HPV）是一种双链DNA病毒，可引起多种人类疾病，包括多种癌症。感染后，两种病毒蛋白E1和E2与细胞因子一起激活病毒复制，并在复制过程中刺激DNA损伤反应（DDR）。E1-E2使用同源重组（HR）来促进DNA复制，但对参与这一过程的宿主因素的理解仍然不完整。以前，我们证明了III类脱乙酰酶SIRT 1，它可以调节HR，被招募到E1-E2复制DNA和调节复制水平。在这里，我们证明了SIRT 1促进E1-E2复制的保真度，并且SIRT 1的缺失导致DNA修复蛋白Werner解旋酶（WRN）向E1-E2复制DNA的招募减少。CRISPR/Cas9编辑表明，WRN与SIRT 1一样，调节E1-E2复制的数量和保真度。这是WRN调节E1-E2 DNA复制的第一份报告，或者WRN在HPV生命周期中的作用。在缺乏SIRT 1的情况下，WRN的乙酰化和稳定性增加，但与E1-E2复制DNA相互作用的能力降低。我们提出了一个模型，其中E1-E2复制打开DDR，刺激SIRT 1去乙酰化WRN。这种去乙酰化促进WRN与E1-E2复制DNA的相互作用，以控制复制的数量和保真度。该系统不仅提供了对HPV复制控制的重要见解，还为研究SIRT 1和WRN在控制哺乳动物细胞复制中的联系提供了独特的模型。重要信息HPV 16是导致全球3%至4%癌症的主要病毒性人类致癌物。感染后，该病毒激活DNA损伤反应（DDR）以促进其生命周期，并将DDR蛋白招募到其复制DNA中，以促进复制期间的同源重组。这促进了有活力的病毒后代的产生。我们对HPV 16复制如何与DDR相互作用的理解仍然不完整。在这里，我们证明了细胞脱乙酰酶SIRT 1，这是E1-E2复制复合物的一部分，调节招聘的DNA修复蛋白WRN的复制DNA。我们证明，WRN调节E1-E2复制的水平和保真度。总的来说，这些结果表明了一种机制，通过这种机制，WRN的SIRT 1脱乙酰化促进了它与E1-E2复制DNA的相互作用，以控制复制的水平和保真度。
Human papillomaviruses (HPV) are double-stranded DNA viruses causative in a host of human diseases, including several cancers. Following infection, two viral proteins, E1 and E2, activate viral replication in association with cellular factors and stimulate the DNA damage response (DDR) during the replication process. E1-E2 uses homologous recombination (HR) to facilitate DNA replication, but an understanding of host factors involved in this process remains incomplete. Previously, we demonstrated that the class III deacetylase SIRT1, which can regulate HR, is recruited to E1-E2-replicating DNA and regulates the level of replication. Here, we demonstrate that SIRT1 promotes the fidelity of E1-E2 replication and that the absence of SIRT1 results in reduced recruitment of the DNA repair protein Werner helicase (WRN) to E1-E2-replicating DNA. CRISPR/Cas9 editing demonstrates that WRN, like SIRT1, regulates the quantity and fidelity of E1-E2 replication. This is the first report of WRN regulation of E1-E2 DNA replication, or a role for WRN in the HPV life cycle. In the absence of SIRT1 there is an increased acetylation and stability of WRN, but a reduced ability to interact with E1-E2-replicating DNA. We present a model in which E1-E2 replication turns on the DDR, stimulating SIRT1 deacetylation of WRN. This deacetylation promotes WRN interaction with E1-E2-replicating DNA to control the quantity and fidelity of replication. As well as offering a crucial insight into HPV replication control, this system offers a unique model for investigating the link between SIRT1 and WRN in controlling replication in mammalian cells.IMPORTANCE HPV16 is the major viral human carcinogen responsible for between 3 and 4% of all cancers worldwide. Following infection, this virus activates the DNA damage response (DDR) to promote its life cycle and recruits DDR proteins to its replicating DNA in order to facilitate homologous recombination during replication. This promotes the production of viable viral progeny. Our understanding of how HPV16 replication interacts with the DDR remains incomplete. Here, we demonstrate that the cellular deacetylase SIRT1, which is a part of the E1-E2 replication complex, regulates recruitment of the DNA repair protein WRN to the replicating DNA. We demonstrate that WRN regulates the level and fidelity of E1-E2 replication. Overall, the results suggest a mechanism by which SIRT1 deacetylation of WRN promotes its interaction with E1-E2-replicating DNA to control the levels and fidelity of that replication.