Association of Human Papillomavirus 16 E2 with Rad50-Interacting Protein 1 Enhances Viral DNA Replication.
Association of Human Papillomavirus 16 E2 with Rad50-Interacting Protein 1 Enhances Viral DNA Replication.
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DOI：
10.1128/jvi.02305-16
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发表时间：
2017-03-01
影响因子：
5.4
通讯作者：
Parish JL
中科院分区：
文献类型：
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作者：
Campos-León K;Wijendra K;Siddiqa A;Pentland I;Feeney KM;Knapman A;Davies R;Androphy EJ;Parish JL
Rad50相互作用蛋白1 （Rint1）与DNA损伤反应蛋白Rad50在S期到G2/M期的过渡过程中相互作用，并在辐射诱导的G2检查点控制中起作用。研究还表明，Rint1通过与Zeste-White 10 （ZW10）的相互作用，在高尔基体到内质网（ER）的囊泡运输中是必不可少的。我们已经分离出一种新的Rint1与人乳头瘤病毒16 （HPV16）转录和复制因子E2之间的相互作用。E2在其ZW10相互作用域中与Rint1结合，我们发现在E2缺失的情况下，Rint1定位于内质网并与ZW10结合。E2表达导致Rint1- zw10相互作用的破坏和核Rint1的积累，与从内质网到高尔基体的囊泡运动显著减少相一致。有趣的是，细胞核Rint1和Mre11/Rad50/Nbs1 （MRN）复合体的成员在不同的E2核病灶中被发现，在s期中期达到峰值，这表明Rint1在细胞核内被募集到E2病灶也可能导致这种DNA损伤感应蛋白复合体的募集。我们发现外源性Rint1表达增强e2依赖性病毒复制。相反，截断的Rint1蛋白的过表达保留E2结合域而不保留Rad50结合域，作为E2依赖性HPV复制的显性阴性抑制剂。综上所述，这些实验表明Rint1和E2之间的相互作用在HPV复制中具有重要作用。HPV感染是许多上皮性癌症的重要驱动因素，包括那些在肛门生殖器和口咽道。人乳头瘤病毒的生命周期受到严格调控，并与其感染的上皮细胞的分化密切相关。在细胞核中形成的HPV复制工厂是复制病毒DNA以支持病毒持续存在和扩大感染的位置。将特定的细胞蛋白复合物招募到这些工厂有助于有效和受控的病毒复制。我们已经确定了一种新的hpv -宿主相互作用，在细胞对DNA损伤和细胞周期控制的反应中起作用。我们发现HPV E2蛋白靶向rad50相互作用蛋白1 （Rint1）以促进病毒基因组复制。这些发现增加了我们对HPV如何复制和宿主细胞途径的理解，这些途径是HPV支持病毒复制的目标。了解这些途径将有助于进一步研究HPV基因组复制的新型抑制剂。
Rad50-interacting protein 1 (Rint1) associates with the DNA damage response protein Rad50 during the transition from the S phase to the G2/M phase and functions in radiation-induced G2 checkpoint control. It has also been demonstrated that Rint1 is essential in vesicle trafficking from the Golgi apparatus to the endoplasmic reticulum (ER) through an interaction with Zeste-White 10 (ZW10). We have isolated a novel interaction between Rint1 and the human papillomavirus 16 (HPV16) transcription and replication factor E2. E2 binds to Rint1 within its ZW10 interaction domain, and we show that in the absence of E2, Rint1 is localized to the ER and associates with ZW10. E2 expression results in a disruption of the Rint1-ZW10 interaction and an accumulation of nuclear Rint1, coincident with a significant reduction in vesicle movement from the ER to the Golgi apparatus. Interestingly, nuclear Rint1 and members of the Mre11/Rad50/Nbs1 (MRN) complex were found in distinct E2 nuclear foci, which peaked during mid-S phase, indicating that the recruitment of Rint1 to E2 foci within the nucleus may also result in the recruitment of this DNA damage-sensing protein complex. We show that exogenous Rint1 expression enhances E2-dependent virus replication. Conversely, the overexpression of a truncated Rint1 protein that retains the E2 binding domain but not the Rad50 binding domain acts as a dominant negative inhibitor of E2-dependent HPV replication. Put together, these experiments demonstrate that the interaction between Rint1 and E2 has an important function in HPV replication. IMPORTANCE HPV infections are an important driver of many epithelial cancers, including those within the anogenital and oropharyngeal tracts. The HPV life cycle is tightly regulated and intimately linked to the differentiation of the epithelial cells that it infects. HPV replication factories formed in the nucleus are locations where viral DNA is copied to support virus persistence and amplification of infection. The recruitment of specific cellular protein complexes to these factories aids efficient and controlled viral replication. We have identified a novel HPV-host interaction that functions in the cellular response to DNA damage and cell cycle control. We show that the HPV E2 protein targets Rad50-interacting protein 1 (Rint1) to facilitate virus genome replication. These findings add to our understanding of how HPV replicates and the host cell pathways that are targeted by HPV to support virus replication. Understanding these pathways will allow further research into novel inhibitors of HPV genome replication.