Interactions of the CA Protein in the Retrovirus Core

逆转录病毒核心中 CA 蛋白的相互作用

• 批准号: 8461643
• 负责人: REBECCA C. CRAVEN
• 金额: $ 22.25万
• 依托单位: PENNSYLVANIA STATE UNIV HERSHEY MED CTR
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-06-01 至 2014-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8461643
• 关键词: Amino Acids; Anti-Retroviral Agents; Antiviral Agents; Biochemical; Biological Assay; Capsid; Capsid Proteins; Cells; Cleaved cell; Complex; Development; Dissection; Distant; Elements; Enzymes; Event; Gagging; Genetic; Genome; Genus Alpharetrovirus; Goals; HIV; Health; Human; In Situ; In Vitro; Infection; Intervention; Kinetics; Knowledge; Laboratories; Life Cycle Stages; Modeling; Molecular; Morphology; Mutagenesis; Mutation; N-terminal; Pathway interactions; Pharmaceutical Preparations; Polyproteins; Positioning Attribute; Process; Proteins; RNA; RNA replication; Research; Resolution; Retroviridae; Ribonucleoproteins; Role; Rous sarcoma virus; Site-Directed Mutagenesis; Staging; Structural Models; Structural Protein; Structure; T-Lymphocyte; Testing; Virion; Virus; Virus Diseases; Virus Replication; Work; assay development; flexibility; gag Gene Products; in vitro Assay; inhibitor/antagonist; interfacial; intermolecular interaction; novel; pathogen; prevent; resistance mechanism; viral RNA

DESCRIPTION (provided by applicant): The CA protein is the major structural protein in the retrovirus core and a contributor to several stages of the virus life-cycle. In the mature virion core, CA forms a protein shell that surrounds the ribonucleoprotein complex of viral RNA and replication enzymes. The integrity of the CA protein and the shell that it forms are critical for the ability of the virus to replicate its genome and to establish infection in the target cell. As such, the CA protein and the maturation process that creates the functional capsid are attractive targets for antiviral intervention. However, the limited knowledge about the precise molecular details of these events seriously limits the ability to exploit these targets. Once CA is cleaved from its polyprotein precursor, CA undergoes a complicated choreography of conformational changes and intermolecular interactions to create the functional capsid. We have developed an in vitro assay for CA assembly that faithfully recapitulates many features of capsid assembly in situ, including the formation of capsid-like shells that contain CA in both hexameric and pentameric arrangements. We have demonstrated that CA assembly in vitro is a well-behaved nucleation-driven process that is highly amenable to the study of the genetic and biochemical factors that control the nucleation of capsid assembly. We have also produced a high resolution structural model for the CA protein in a pentameric arrangement that both identifies novel intermolecular interactions between the CA subunits and provides a testable model for the control of capsid nucleation. In the proposed studies we will test key features of this model, including the action of particular interfacial residues, of the ?8 helix and the ?8-??9 loop, and of the ?-hairpin and flexible outer loop regions of the N-terminal domain in controlling the nucleation of capsid assembly. We will seek to define CA intersubunit interactions in capsid assembly intermediates and the role of other virion constituents in the process. Achieving these goals will be a major step toward understanding one of the most mysterious aspects of the retroviral life cycle and the ultimate identification of new strategies for blocking the maturation events and thereby the replication of the virus.

描述（由申请人提供）：CA蛋白是逆转录病毒核心的主要结构蛋白，也是病毒生命周期的多个阶段的贡献者。在成熟的病毒素内核中，Ca形成了一个蛋白质壳，该蛋白壳围绕病毒RNA和复制酶的核糖核蛋白复合物。 CA蛋白及其形成的壳的完整性对于病毒复制其基因组并在靶细胞中建立感染的能力至关重要。因此，创建功能性capsid的CA蛋白和成熟过程是抗病毒干预的有吸引力的靶标。但是，关于这些事件的确切分子细节的有限知识严重限制了利用这些目标的能力。一旦Ca从其多蛋白前体裂开后，CA就会经历复杂的构象变化和分子间相互作用的编排，以创建功能性上限。我们已经开发了一种用于CA组件的体外测定法，该测定法忠实地概括了原位带帽的许多特征，包括形成带有六聚体和五聚体布置的ca的带状壳样壳。我们已经证明，在体外CA组装是一种表现良好的成核驱动的过程，它高度适合研究控制衣壳组装成核的遗传和生化因子。我们还以五聚体排列产生了CA蛋白的高分辨率结构模型，这两者都识别了CA亚基之间的新型分子间相互作用，并为控制衣壳成核的控制提供了可测试的模型。在拟议的研究中，我们将测试该模型的关键特征，包括？8螺旋的特定界面残基的作用，以及？8-- ?? 9回路，以及？ - 海 - 海p和柔性外圈区域的柔性外环区域，以控制capsid组件的成核。我们将寻求定义CAPSID组装中间体中的CA间相互作用以及其他病毒素成分在此过程中的作用。实现这些目标将是理解逆转录病毒生命周期中最神秘的方面之一，以及对阻止成熟事件的新策略的最终识别，从而复制病毒的最终识别。

项目成果

Lethal mutations in the major homology region and their suppressors act by modulating the dimerization of the rous sarcoma virus capsid protein C-terminal domain.

主要同源区及其抑制子中的致死突变通过调节劳斯肉瘤病毒衣壳蛋白 C 末端结构域的二聚化起作用。

• DOI: 10.1002/prot.24188
• 发表时间: 2013
• 期刊: Proteins
• 影响因子: 2.9
• 作者: Dalessio,PaulaM;Craven,RebeccaC;Lokhandwala,ParvezM;Ropson,IraJ
• 通讯作者: Ropson,IraJ

Retroviral capsid assembly: a role for the CA dimer in initiation.

• DOI: 10.1016/j.jmb.2009.04.006
• 发表时间: 2009-06-05
• 期刊: JOURNAL OF MOLECULAR BIOLOGY
• 影响因子: 5.6
• 作者: Purdy, John G.;Flanagan, John M.;Ropson, Ira J.;Craven, Rebecca C.
• 通讯作者: Craven, Rebecca C.

Visualization of a missing link in retrovirus capsid assembly.

• DOI: 10.1038/nature07724
• 发表时间: 2009-02-05
• 期刊: NATURE
• 影响因子: 64.8
• 作者: Cardone, Giovanni;Purdy, John G.;Cheng, Naiqian;Craven, Rebecca C.;Steven, Alasdair C.
• 通讯作者: Steven, Alasdair C.