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Molecular Determinants of Retroviral Capsid Morphology using a Novel Imaging Approach

使用新型成像方法研究逆转录病毒衣壳形态的分子决定因素

基本信息

批准号：
10219047
负责人：
Nathaniel Lee Talledge
金额：
$ 6.88万
依托单位：
UNIVERSITY OF MINNESOTA
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-03-17 至 2022-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10219047
关键词：
3-Dimensional Alpha Particles Anti-Retroviral Agents Architecture Benchmarking Biogenesis Biological Biophysics Capsid Capsid Proteins Cell membrane Cells Complex Core Protein Crowding Cryo-electron tomography Cryoelectron Microscopy Disease Drug Design Drug Targeting Electron Microscopy Elements Foundations Goals HIV HIV-1 HIV-2 Health Human Human T-lymphotropic virus 1 Hybrids Image Individual Knowledge Life Cycle Stages Location Maps Mediating Methodology Methods Modeling Molecular Molecular Conformation Morphology Nature Outcome Pathway interactions Peptides Pharmacotherapy Positioning Attribute Process Production Property Proteins Radiation induced damage Research Resolution Retroviridae Sampling Structure Techniques Testing Tomogram Tube Ursidae Family Viral Virion Virus Virus Assembly Virus Diseases Virus-like particle antiretroviral therapy base comparative drug development electron radiation flexibility gag Gene Products image processing imaging approach insight member new therapeutic target novel novel strategies particle reconstruction targeted treatment tool

项目摘要

Project Summary Immature virus particle structure can provide valuable clues to the nature of virus particle assembly in cells. In this proposal, we sought to decipher key structural details of immature retroviral Gag assemblies by obtaining high-resolution structures using cryo-electron microscopy (cryo-EM). Reconstructions from the immature Gag structure of distinct types of virus-like particles reveals distinct similarities among Gag lattice organization of the viruses studied to date. Structural insights have enhanced from a comparative analysis human immunodeficiency virus type 1 (HIV-1) cryo-EM reconstructions regarding the importance of the capsid (CA) carboxy-terminal domain (CTD) and spacer peptide 1 (SP1) regions in forming hexameric assemblies of CA in the intermolecular contacts of the overall lattice structure. In order to gain a fundamental understanding of retrovirus assembly and maturation, atomic resolution maps of immature particles for other members of Orthoretrovirinae are required. Here, I propose a strategy, i.e., cryo-electron tomography guided single particle reconstruction (Cryo-ET guided SPR), to achieve atomic resolution reconstruction maps of immature retrovirus particles. Initial efforts at streamlining the EM methodology to resolve immature Gag lattices using the strengths from each imaging processing technique combined into a new approach. I will apply this methodology to study HIV type 2 (HIV-2) and human T-cell leukemia virus type 1 (HTLV-1). HTLV-1 has an immature Gag lattice that forms a uniquely flattened capsid that is not observed in other retroviruses morphologies, such as with HIV-1. My central hypothesis is that HTLV-1 Gag CA protein mediates unique intermolecular contacts in the immature Gag lattice that allows hexameric Gag conformations to generate a flattened morphology that does not exist within immature HIV viral particles. By studying the CA structures of HIV-2 and HTLV-1, I expect to generate biophysical and structural information to provide fundamental aspects of virus particle assembly and Gag architecture. To expand the biological significance of this proposed research morphological aspects of authentic HIV-2 and HTLV-1 immature virus particle capsid lattices will be analyzed and compaired to the more ordered viral-like immature particles used for high-resolution efforts. A single particle and tomography cryo-EM hybrid approach of both capsid lattices will provide novel insights into this flattened CA morphology as well as the conformational flexibility of Gag in forming immature capsid structures. Identifying structural differences in the Gag lattice organization among closely related human retroviruses sheds new details on virus particle assembly that will ultimately identify novel drug targets for antiretroviral therapies targeting Gag capsid lattice formation.

项目总结