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RNA binding and packaging by retroviral Gag proteins

逆转录病毒 Gag 蛋白的 RNA 结合和包装

基本信息

批准号：
10034983
负责人：
Karin M Musier-Forsyth
金额：
$ 51.53万
依托单位：
OHIO STATE UNIVERSITY
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 2025-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10034983
关键词：
5&apos Untranslated Regions Acylation Adopted Affect Binding Binding Sites Biological Assay Biology Cell Culture Techniques Cells Characteristics Clinical Computer Models Data Drug Targeting Elements Ensure Fluorescence Resonance Energy Transfer Genetic Materials HIV-1 Heterogeneity Hydroxyl Radical In Vitro Kinetics Label Mass Spectrum Analysis Measures Mediating Molecular Conformation Mutation Nucleotides Pharmaceutical Preparations Primer Extension Production Property RNA RNA Binding RNA Conformation RNA Probes RNA Splicing Reporting Role Specificity Structure Testing Therapeutic Therapeutic Agents Transcript Transcription Initiation Site Viral Viral Genome Virion Virus Assembly Virus Replication Work Zinc Fingers base cost crosslink design dimer dimethyl sulfate expectation fitness gag Gene Products genomic RNA hammerhead ribozyme inhibitor/antagonist insight mutant novel strategies novel therapeutics particle preference prevent single molecule stoichiometry targeted treatment viral RNA

项目摘要

Abstract HIV-1 packages two copies of genomic RNA (gRNA) as a dimer into newly formed viral particles. Retroviral assembly doesn’t depend on the presence of gRNA, yet gRNA packaging is essential for production of infectious virus particles. Robust and specific mechanisms must therefore be in place to ensure the genetic material is passed on to progeny virions. While there are numerous therapeutics in clinical use, there are currently no gRNA packaging or viral assembly inhibitors. Selective gRNA packaging is facilitated by interactions between the HIV- 1 Gag protein and conserved gRNA elements within the 5′ untranslated region (5′UTR). Here, we focus on recent unexpected findings in HIV-1 RNA biology that revealed sequence heterogeneity at the 5′ end of the HIV-1 RNA transcript. The variable number of G residues (1G, 2G and 3G) has been reported to affect the gRNA localization, with 1G RNA preferentially selected over 3G to be the viral genome. This is very surprising given that these two 9-kb RNAs only differ by 2 nucleotides. Preliminary data presented here strongly support the enrichment of 1G- containing transcripts among packaged gRNA. Importantly, various point mutants that abrogate the preference for 1G RNA have been identified. Exciting preliminary data support our major hypothesis that the ensemble of RNA structures adopted by the 5′UTR is significantly altered in 1G vs. 3G transcripts. These data provide a strong starting point for the proposed studies aimed at elucidating the mechanism by which transcriptional start site choice modulates gRNA packaging selectivity. More broadly, we will gain insights into how subtle sequence changes can alter the ensemble of 5′UTR RNA structures and impact viral replication fitness. The specific aims are: (1) To probe wild-type and mutant retroviral gRNA structure and dynamics; (2) To probe wild-type and mutant HIV-1 Gag RNA binding and packaging specificity. The results of these studies will help guide the design of novel therapeutic agents that target the 5′UTR and interfere with the essential conformational plasticity and/or key binding interactions with the expectation for a lower rate of mutational escape than conventional drugs.

摘要