CRYSTALLOGRAPHIC ANALYSIS OF HIV-1 VIF IN COMPLEX WITH HUMAN HOST PROTEINS

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 世界卫生组织估计，全世界有3000-3600万人感染了HIV-1病毒。自1981年以来，几乎有同样多的人死于艾滋病。尽管在开发新的抗病毒疗法方面取得了重大进展，但由于耐药性的出现，需要新的药物类别。在这里，我们建议确定HIV-1因子Vif的晶体结构，由于它能够灭活CD4+T细胞中的天然免疫因子APOBEC3G，因此对于病毒的感染性是必不可少的。VIF通过将APOBEC3G募集到库林环连合酶(CRL)复合体中发挥作用，在CRL复合体中，抗病毒因子经历多泛素化，随后被26S蛋白酶体降解。Vif的C端72个氨基酸是与CRL的细长蛋白(Elo)B和EloC蛋白通过发散的BC-box基序相互作用的必要条件和充分条件。我们已经制备了一个由Vif的C末端与人EloB/C蛋白结合的三元复合体的晶体。在短期内，我们需要快速获得同步辐射，以：(I)筛选我们在国内制备的大量小晶体，以指导优化工作；以及(Ii)收集使用已知的EloB/C坐标进行可能的分子替换的本地数据。Vif和EloB/C之间的相互作用代表着新的HIV-1疗法的潜在新靶点。因此，这项工作有可能造福公众健康。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The World Health Organization estimates 30-36 million persons are infected worldwide with the HIV-1 virus. Nearly as many individuals have died from AIDS since 1981. Although significant progress has been made in the development of new anti-viral therapies, new classes of drugs are required due to the emergence of drug resistance. Here we propose to determine the crystal structure of the HIV-1 factor Vif, which is essential for viral infectivity due to its ability to inactivate the innate immune factor APOBEC3G in CD4+ T cells. Vif functions in the infected host cell by recruiting APOBEC3G to a Cullin Ring Ligase (CRL) complex where the anti-viral factor undergoes polyubiquitination, and subsequent degradation by the 26S proteasome. The C-terminal 72 amino acids of Vif are necessary and sufficient to interact with the elongin (Elo)B and EloC proteins of the CRL via a divergent BC-box motif. We have prepared crystals of a ternary complex comprising the C-terminus of Vif bound to the human EloB/C proteins. In the short term, we require rapid access to synchrotron radiation to: (i) screen the numerous small crystals we have prepared at home in order to guide optimization efforts; and (ii) to collect native data for possible molecular replacement using the known EloB/C coordinates. The interaction between Vif and EloB/C represents a potentially novel target for new HIV-1 therapeutics. As such, this work has the potential to benefit public health.