Lampel Ayala, Bram Yaron, Ezer Anat, Shaltiel-Kario Ronit, Saad Jamil S, Bacharach Eran, Gazit Ehud
†Department of Molecular Microbiology and Biotechnology, George S. Wise Faculty of Life Sciences, Tel Aviv University, Tel Aviv 69978, Israel.
‡Department of Microbiology, University of Alabama at Birmingham, Birmingham, Alabama 35294, United States.
ACS Chem Biol. 2015 Aug 21;10(8):1785-90. doi: 10.1021/acschembio.5b00347. Epub 2015 May 29.
Viral assembly, similar to other self-organizing protein systems, relies upon early building blocks, which associate into the late supramolecular structures. An initial and crucial event during HIV-1 core assembly is the dimerization of the capsid protein C-terminal domain, which stabilizes the viral capsid lattice. Thus, monitoring and manipulating this stage is desirable both from mechanistic as well as clinical perspectives. Here, we developed a fluorescent-based method for the detection and visualization of these early capsid interactions. We detected strong dimeric interactions, which were influenced by mutations in the capsid protein. We utilized this assay for potential assembly inhibitors screening, which resulted in the identification of a leading compound that hinders the assembly of capsid protein in vitro. Moreover, a derivative of the compound impaired virus production and infectivity in cell cultures. These findings demonstrate that the described assay efficiently detects the very first association events in HIV-1 capsid formation and emphasize the significance of targeting early intermolecular interactions.
病毒组装与其他自组织蛋白质系统类似，依赖于早期构建模块，这些模块会组装成晚期超分子结构。HIV-1核心组装过程中的一个初始且关键的事件是衣壳蛋白C末端结构域的二聚化，这使病毒衣壳晶格得以稳定。因此，从机制和临床角度来看，监测和操控这一阶段都是很有必要的。在此，我们开发了一种基于荧光的方法来检测和可视化这些早期衣壳相互作用。我们检测到了强烈的二聚体相互作用，这种相互作用受到衣壳蛋白突变的影响。我们利用该检测方法进行潜在组装抑制剂的筛选，结果鉴定出一种先导化合物，该化合物在体外可阻碍衣壳蛋白的组装。此外，该化合物的一种衍生物会损害细胞培养中的病毒产生和感染性。这些发现表明，所描述的检测方法能够有效检测HIV-1衣壳形成过程中最初的缔合事件，并强调了靶向早期分子间相互作用的重要性。
