STRUCT ANALYSIS OF THE PROTEIN NETWORK OF SORTING AT MULTIVESICULAR BODIES:HIV

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 HIV-1颗粒的组装和释放依赖于人蛋白AIP 1/阿利克斯和Vps 4A/B，以及四种人蛋白复合物：Hrs/STAM和ESCRT-I、II和III。这些蛋白质和复合物从酵母到人类都是保守的，它们的正常功能是将单泛素化受体、酶和其他货物分类到溶酶体或液泡。在这个网络中测试的几种蛋白质中的任何一种失活都会阻止HIV的释放和感染性。我们已经开始系统地确定该网络中的结构域、蛋白质和多蛋白复合物的结构，并且我们已经解决了Vps 27（酵母Hrs）FYVE结构域、Bro 1的N-末端半部分（酵母AIP 1/阿利克斯）和几乎完整的酵母ESCRT-II复合物（Vps 22、Vps 25和Vps 36）的结构。在接下来的两年里，我们计划1）解决ESCRT-I的结构（Vps 23，Vps 28，Vps 37）复合物的结构和功能分析; 2）以我们的3.6复合物结构为出发点，对ESCRT-Ⅱ进行结构和功能分析; 3）解析Bro 1或AIP 1/阿利克斯的C-末端一半的结构;和4）表征AIP 1/阿利克斯的膜和病毒晚期结构域结合。从长远来看，我们的目标是对整个网络进行完整的结构和生化表征。

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. HIV-1 particle assembly and release depend on the human proteins AIP1/Alix and Vps4A/B, and four human protein complexes: Hrs/STAM and ESCRT-I, II, and III. These proteins and complexes are conserved from yeast to human, and their normal function is to sort monoubiquitinated receptors, enzymes, and other cargo to the lysosome or vacuole. Inactivation of any one of several proteins tested in this network blocks HIV release and infectivity. We have begun to systematically determine the structures of domains, proteins, and multiprotein complexes in this network, and we have already solved the structures of the Vps27 (yeast Hrs) FYVE domain, the N-terminal half of Bro1 (yeast AIP1/Alix), and the nearly intact yeast ESCRT-II complex (Vps22,Vps25, and Vps36). Over the next two years we plan to 1) solve the structure of the ESCRT-I (Vps23, Vps28, Vps37) complex from yeast or human; 2) carry out a structural and functional analysis of ESCRT-II taking our 3.6 ¿ complex structure as a starting point; 3) solve the structure of the C-terminal half of Bro1 or AIP1/Alix; and 4) characterize the membrane and viral late domain binding of AIP1/Alix. In the long term our goal is the complete structural and biochemical characterization of the entire network.