Molecular and cellular determinants of TRIM5alpha restriction of HIV-1

摘要 TRIM 5 α是一种限制性因子，在感染过程中靶向逆转录病毒衣壳，通过以下方式抑制感染： 诱导病毒衣壳核心的解体失败。发生这种情况的机制是很差的 明白我们提供的数据表明，TRIM5α的动态构象变化与 抑制病毒感染的能力，并建议更好地定义这些构象变化，以了解 分子间的相互作用驱动衣壳解体。在目标1中，我们将定义构象变化， 发生在恒河猴TRIM5α、人类TRIM5α和一组天然发生的和结构引导的突变中 为了定义在重组体的背景下驱动这些构象变化的分子相互作用， TRIM5α二聚体单元，包含卷曲螺旋结构域和接头2区。在目标2中，我们将扩展这些 研究确定这些构象变化如何翻译到相邻的结构域，包括衣壳 结合TRIM5α的SPRY结构域，并确定SPRY与组装CA的结合如何影响这些 构象变化在目标3中，我们提出了在前2个目标中获得的结果的功能验证， 这些实验将确定生物物理和生物化学蛋白质如何转化为 在限制过程中执行单独的、可衡量的步骤。

Abstract TRIM5α is a restriction factor which targets the retroviral capsid during infection which inhibits infection by inducing the abortive disassembly of the viral capsid core. The mechanism by which this occurs is poorly understood. We provide data demonstrating that dynamic conformational changes in TRIM5α correlate with the ability to inhibit viral infection, and propose to better define these conformational changes to understand the molecular interactions that drive capsid disassembly. In aim 1, we will define the conformational changes that occur in rhesus TRIM5α, human TRIM5α, and a panel of naturally occurring and structurally guided mutations to define the molecular interactions that drive these conformational changes in the context of the recombinant TRIM5α dimeric unit comprising the coiled coil domain and linker 2 region. In Aim 2, we will expand these studies to define how these conformational changes translate to neighboring domains, including the capsid binding SPRY domain of TRIM5α, and also determine how SPRY binding to assembled CA influences these conformational changes. In aim 3, we propose functional validation of the results obtained in the first 2 aims in experiments which will determine how the biophysical and biochemical proteins translate to the ability to perform the individual, measurable steps in the restriction process.