THE EFFECTS OF DOMAIN SWAPPING IN HIV-1 CAPSID PROTEIN

HIV-1 衣壳蛋白结构域交换的影响

• 批准号: 8168736
• 负责人: Peter E. Prevelige
• 金额: $ 0.93万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-03-10 至 2010-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8168736
• 关键词: Amino Acids; C-terminal; Capsid; Capsid Proteins; Computer Retrieval of Information on Scientific Projects Database; DNA Sequence Rearrangement; Dimerization; Elements; Engineering; Funding; Gambling; Grant; HIV; HIV-1; Hydrogen Bonding; Individual; Institution; Mutation; N-terminal; Play; Process; Research; Research Personnel; Resources; Retroviridae; Role; Scanning; Solutions; Source; Structural Models; Structural Protein; United States National Institutes of Health; Viral; Virion; Zinc Fingers; dimer; flexibility; oxidation

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 assembly of HIV-1 is a multi-step process in which several individual structural proteins undergo a substantial morphological rearrangement. The capsid (CA) protein plays a crucial but poorly understood role in this assembly. CA contains two-independently folded domains, the N-terminal (NTD) and C-terminal (CTD) domains, connected by a flexible linker. The CTD of HIV-1 has been shown to dimerize in solution both in the mature capsid protein and in the isolated domain (Gamble et al. 1997). The current structural model of the mature virion indicates the fundamental building block of the mature core is attributed to a CA hexamer formed by the self-association of the NTD tied together by dimerization of the CTD (Gamble et al. 1997). Additionally, CTD dimerization has been shown to be required for this hexamer formation (Lanman et al. 2003). The CTD of HIV contains the major homology region (MHR), a sequence of 20 amino acids that is highly conserved across different genera of retroviruses (Wills and Craven 1991). It is well documented that viral assembly is highly sensitive to MHR mutations but from a structural perspective it is not clear why MHR mutations are so deleterious (Cairns and Craven 2001). Recent studies have shown that a structural homology exists between the CTD of CA and the dimeric zinc finger associated domain SCAN (Ivanov et al. 2005). However, the SCAN domain dimer is domain swapped in comparison to the CTD of CA. In a domain swapped dimer, a structural element from one subunit is exchanged with the corresponding structural element from another subunit (Liu and Eisenberg 2002). The domain swapped region of the SCAN dimer corresponds to the MHR of the CTD. A domain swapped dimer would provide an explanation for the importance of the MHR because hydrogen bonding in this region would occur across the dimer interface. To determine whether CA can assemble when the C-terminal domain is domain swapped a fusion construct was engineered where the C-terminal domain of HIV-1 CA was replaced with the SCAN domain. These interactions will be studied using H/D exchange and fast photochemical oxidation.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 HIV-1的组装是一个多步骤过程，其中几种单独的结构蛋白会经历大量的形态重排。衣壳（CA）蛋白在该组装中起着至关重要但知之甚少的作用。 CA包含由柔性接头连接的两种独立折叠域，即N末端（NTD）和C末端（CTD）域。 HIV-1的CTD已显示在成熟的衣壳蛋白和分离的结构域中的溶液中均匀化（Gamble等，1997）。 成熟病毒体的当前结构模型表明，成熟核心的基本构建块归因于通过CTD的二聚化将NTD的自我关联形成的Ca Hexamer（Gamble等，1997）。 另外，已经证明CTD二聚化是这种己酯形成所必需的（Lanman等，2003）。 HIV的CTD包含主要同源区域（MHR），这是20种氨基酸的序列，在不同的逆转录病毒的不同属中高度保守（Wills and Craven 1991）。 有充分的文献证明，病毒组装对MHR突变高度敏感，但是从结构的角度来看，尚不清楚为什么MHR突变如此有害（Cairns and Craven 2001）。最近的研究表明，CA和二聚体锌指相关结构域扫描之间存在结构同源性（Ivanov等，2005）。 但是，与CA的CTD相比，扫描域二聚体是换的。在域交换二聚体中，一个亚基的结构元件与另一个亚基的相应结构元件交换（Liu and Eisenberg 2002）。 扫描二聚体的域交换区域对应于CTD的MHR。域交换二聚体将为MHR的重要性提供解释，因为该区域中的氢键将在二聚体界面上发生。为了确定当C末端结构域是域交换时CA是否可以组装一个融合构建体，在将HIV-1 Ca的C末端结构域用扫描域代替。 这些相互作用将使用H/D交换和快速的光化学氧化研究。