Host proteins interacting with the Moloney murine leukemia virus integrase: multiple transcriptional regulators and chromatin binding factors.
Host proteins interacting with the Moloney murine leukemia virus integrase: multiple transcriptional regulators and chromatin binding factors.
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DOI：
10.1186/1742-4690-5-48
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发表时间：
2008-06-13
期刊：
影响因子：
3.3
通讯作者：
Goff SP
中科院分区：
文献类型：
--
作者：
Studamire B;Goff SP
逆转录病毒复制的关键步骤是将前病毒稳定整合到其宿主的基因组中。病毒整合酶蛋白在逆转录病毒生命周期的这一重要步骤中至关重要。虽然哺乳动物逆转录病毒整合的基本机制已经得到很好的表征，但决定病毒整合事件如何靶向基因组的特定区域或特定DNA结构的区域的因素仍然定义不清。关于宿主蛋白对靶位点的选择、整合中间体的修复以及整合效率的影响，仍然存在重大问题。我们描述了酵母双杂交筛选的结果，使用莫洛尼鼠白血病病毒整合酶作为诱饵，筛选与整合酶相互作用的宿主蛋白质的小鼠cDNA文库。我们鉴定了27种与不同整合酶融合蛋白相互作用的蛋白质。鉴定的蛋白质包括染色质重塑、DNA修复和转录因子（13种蛋白质）;翻译调节因子、解旋酶、剪接因子和其他RNA结合蛋白（10种蛋白质）;以及转运蛋白或杂项因子（4种蛋白质）。我们证实了这些蛋白质与整合酶的相互作用，通过测试他们在其他酵母菌株与GAL 4-DNA结合结构域整合酶融合的背景下，并通过重组蛋白之间的体外结合试验。随后的分析表明，许多被鉴定为Mo-MLV整合酶相互作用因子的蛋白质也在酵母和体外与HIV-1整合酶相互作用。我们确定了几种蛋白质直接相互作用的MoMLV和HIV-1整合酶，这可能是共同的整合反应途径的两种病毒。在屏幕上识别的许多蛋白质是整合酶的逻辑相互作用伙伴，并且许多相互作用的有效性得到了其他研究的支持。此外，我们观察到一些蛋白质已经记录了与其他病毒的相互作用，提出了一种有趣的可能性，即不同病毒可能使用共同的宿主蛋白质。我们进行了这个屏幕，以确定主机的因素，可能会影响整合靶位点的选择，并发现我们的屏幕产生了丰富的假定相互作用的蛋白质，值得进一步研究。
A critical step for retroviral replication is the stable integration of the provirus into the genome of its host. The viral integrase protein is key in this essential step of the retroviral life cycle. Although the basic mechanism of integration by mammalian retroviruses has been well characterized, the factors determining how viral integration events are targeted to particular regions of the genome or to regions of a particular DNA structure remain poorly defined. Significant questions remain regarding the influence of host proteins on the selection of target sites, on the repair of integration intermediates, and on the efficiency of integration. We describe the results of a yeast two-hybrid screen using Moloney murine leukemia virus integrase as bait to screen murine cDNA libraries for host proteins that interact with the integrase. We identified 27 proteins that interacted with different integrase fusion proteins. The identified proteins include chromatin remodeling, DNA repair and transcription factors (13 proteins); translational regulation factors, helicases, splicing factors and other RNA binding proteins (10 proteins); and transporters or miscellaneous factors (4 proteins). We confirmed the interaction of these proteins with integrase by testing them in the context of other yeast strains with GAL4-DNA binding domain-integrase fusions, and by in vitro binding assays between recombinant proteins. Subsequent analyses revealed that a number of the proteins identified as Mo-MLV integrase interactors also interact with HIV-1 integrase both in yeast and in vitro. We identify several proteins interacting directly with both MoMLV and HIV-1 integrases that may be common to the integration reaction pathways of both viruses. Many of the proteins identified in the screen are logical interaction partners for integrase, and the validity of a number of the interactions are supported by other studies. In addition, we observe that some of the proteins have documented interactions with other viruses, raising the intriguing possibility that there may be common host proteins used by different viruses. We undertook this screen to identify host factors that might affect integration target site selection, and find that our screens have generated a wealth of putative interacting proteins that merit further investigation.
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