HIV Frameshift Site RNA Structure, Function and Ligand Interactions

描述（由申请人提供）：拟议研究的目的是了解HIV-1移码（FS）机制，并了解如何用小的药物样分子靶向它。HIV-1的FS位点是一个高度保守的基因组RNA结构，位于gag和pol阅读框之间。其目的是刺激Pol基因表达所需的-1 FS， Pol基因位于相对于Gag的-1阅读框中。我们已经解决了HIV-1 FS RNA的结构，无论是单独的还是与小药物样分子的复合物。我们将在体外和体内使用高通量筛选来鉴定结合和调节HIV-1 FS RNA的小分子。最好的小分子配体将被鉴定为先导化合物。HIV-1 FS RNA与先导化合物复合物的结构将得到解决，以了解其作用模式的基础。根据这些信息，新的第二代化合物将被开发出具有更好的亲和力、特异性和效力。此外，我们将研究FS机制，这是目前尚不清楚。阐明FS机制将有助于更好地理解病毒生命周期的这一基本特征，这是所有逆转录病毒所利用的。最后，我们将研究整个HIV-1基因组FS结构域的结构和功能，该结构域最近被发现，并被假设在FS之前减缓翻译动力学。

DESCRIPTION (provided by applicant): The purpose of the proposed studies is to understand the HIV-1 frameshift (FS) mechanism and to learn how it can be targeted with small, drug-like molecules. The HIV-1 FS site is a highly conserved genomic RNA structure located between the gag and pol reading frames. Its purpose is to stimulate a -1 FS that is required for expression of the Pol genes, which are in the -1 reading frame relative to Gag. We have solved the structure of the HIV-1 FS RNA, both alone and in complex with small drug-like molecules. We will use high throughput screens to identify small molecules that bind to and modulate the HIV-1 FS RNA, both in vitro and in vivo. The best small molecule ligands will be identified as lead compounds. The structures of the HIV-1 FS RNA in complex with lead compounds will be solved, in order to understand the basis for their modes of action. From this information, novel second generation compounds will be developed with improved affinity, specificity and potency. Additionally, we will investigate the FS mechanism, which is currently not well understood. Elucidation of the FS mechanism will provide a better understanding of this essential feature of the viral life cycle, which is utilized by all retroviruses. Finally, we will investigate the structure and function of the entire HIV-1 genomic FS domain, which has been recently identified and is hypothesized to function in slowing the kinetics of translation prior to the FS. PUBLIC HEALTH RELEVANCE: More than 30 million people are infected with Human immunodeficiency virus (HIV), the causative agent of AIDS. Results of the proposed studies will elucidate how HIV stimulates translational frameshifting, a critical step in the viral replication cycle. Small drug-like molecules that inhibit viral replication by targeting this mechanism will be developed.