Department of Biological Sciences, University of Texas at El Paso, El Paso, Texas, USA.
J Virol. 2013 Mar;87(5):2496-507. doi: 10.1128/JVI.01668-12. Epub 2012 Dec 19.
Poly(ADP-ribose) polymerase 1 (PARP-1) is a cellular enzyme with a fundamental role in DNA repair and the regulation of chromatin structure, processes involved in the cellular response to retroviral DNA integration. However, the function of PARP-1 in retroviral DNA integration is controversial, probably due to the functional redundancy of the PARP family in mammalian cells. We evaluated the function of PARP-1 in retroviral infection using the chicken B lymphoblastoid cell line DT40. These cells lack significant PARP-1 functional redundancy and efficiently support the postentry early events of the mammalian-retrovirus replication cycle. We observed that DT40 PARP-1(-/-) cells were 9- and 6-fold more susceptible to infection by human immunodeficiency virus type 1 (HIV-1)- and murine leukemia virus (MLV)-derived viral vectors, respectively, than cells expressing PARP-1. Production of avian Rous-associated virus type 1 was also impaired by PARP-1. However, the susceptibilities of these cell lines to infection by the nonretrovirus vesicular stomatitis virus were indistinguishable. Real-time PCR analysis of the HIV-1 life cycle demonstrated that PARP-1 did not impair reverse transcription, nuclear import of the preintegration complex, or viral DNA integration, suggesting that PARP-1 regulates a postintegration step. In support of this hypothesis, pharmacological inhibition of the epigenetic mechanism of transcriptional silencing increased retroviral expression in PARP-1-expressing cells, suppressing the differences observed. Further analysis of the implicated molecular mechanism indicated that PARP-1-mediated retroviral silencing requires the C-terminal region, but not the enzymatic activity, of the protein. In sum, our data indicate a novel role of PARP-1 in the transcriptional repression of integrated retroviruses.
聚(ADP-核糖)聚合酶 1(PARP-1)是一种在 DNA 修复和染色质结构调节中具有基本作用的细胞酶,这些过程涉及细胞对逆转录病毒 DNA 整合的反应。然而,PARP-1 在逆转录病毒 DNA 整合中的功能存在争议,这可能是由于哺乳动物细胞中 PARP 家族的功能冗余。我们使用鸡 B 淋巴母细胞系 DT40 评估了 PARP-1 在逆转录病毒感染中的功能。这些细胞缺乏明显的 PARP-1 功能冗余,并且能够有效地支持哺乳动物 - 逆转录病毒复制周期的进入后早期事件。我们观察到,与表达 PARP-1 的细胞相比,DT40 PARP-1(-/-)细胞对人类免疫缺陷病毒 1(HIV-1)和鼠白血病病毒(MLV)衍生的病毒载体的感染分别敏感 9 倍和 6 倍。禽 Rous 相关病毒 1 的产生也受到 PARP-1 的损害。然而,这些细胞系对非逆转录病毒水疱性口炎病毒的感染敏感性无法区分。HIV-1 生命周期的实时 PCR 分析表明,PARP-1 不会损害逆转录、预整合复合物的核输入或病毒 DNA 整合,表明 PARP-1 调节整合后步骤。支持这一假设,对转录沉默的表观遗传机制的药理学抑制增加了表达 PARP-1 的细胞中的逆转录病毒表达,抑制了观察到的差异。对所涉及的分子机制的进一步分析表明,PARP-1 介导的逆转录病毒沉默需要蛋白质的 C 末端区域,但不需要酶活性。总之,我们的数据表明 PARP-1 在整合的逆转录病毒的转录抑制中具有新的作用。