权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Structural Control of Human Co-factors for Retroviral Gene Expression

逆转录病毒基因表达的人类辅因子的结构控制

基本信息

批准号：
9341781
负责人：
CLARA KIELKOPF
金额：
$ 8.45万
依托单位：
UNIVERSITY OF ROCHESTER
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-03-01 至 2020-02-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9341781
关键词：
Address Affinity Alternative Splicing Binding Binding Sites Biochemical Biochemistry Biological Assay Biology Calorimetry Cell Extracts Cell physiology Cells Collaborations Complex Crystallization Development Drug resistance Gene Expression Genome Genomics HIV HIV-1 Hand Health Homologous Gene Housing Human Human Cell Line Immune Infection Integration Host Factors Knowledge Laboratories Length Life Cycle Stages Ligands Mediating Medicine Messenger RNA Molecular Molecular Biology Mutation Nucleocapsid Proteins Pathway interactions Pharmaceutical Preparations Phase Plague Process Production Proteins RNA RNA Binding RNA Recognition Motif RNA Splicing Resistance Resolution Retroviridae Retrovirology Role SF1 Small Interfering RNA Specificity Spliceosomes Staging Structural Biochemistry Structure Surface Testing Therapeutic Therapeutic Intervention Titrations Transcript U2 Small Nuclear Ribonucleoprotein U2 small nuclear RNA Universities Ursidae Family Variant Viral Virus Virus Replication Work X-Ray Crystallography Yeasts base biochemical tools cofactor genomic RNA insight interdisciplinary approach knock-down mRNA Precursor macromolecule medical schools novel novel therapeutic intervention protein expression research study rev Protein small hairpin RNA small molecule stem structural biology therapeutic development virology virus host interaction

项目摘要

DESCRIPTION (provided by applicant): In this project, we will dissect the activity of key host cofactor Tat-SF1 in human versus HIV-1 RNA splicing. Complex retroviruses such as HIV-1 co-opt the human spliceosome machinery for tightly coordinated production of their spliced mRNAs and genomic RNAs during the early and late stages of the viral life cycle. At present, the molecular mechanisms responsible for the dominant progression of HIV-1 through the host splicing cycle remain outstanding questions in the field. Tat-SF1 is a host protein that is critica in the production of fully spliced HIV-1 mRNAs in the early stage of the retroviral life cycle. Tat SF1 is known to associate with the U2 small nuclear ribonucleoprotein subunit (snRNP) of the spliceosome and regulate the splicing of specific human transcripts. As such, studies of Tat-SF1 will be important to understand host-virus interactions, thereby laying the groundwork for the development of new biochemical tools or therapeutic intervention approaches at the level of splicing. At present, the underlying structural and functional roles of Tat-SF1 in HIV-1 and human splicing remain unknown. We will address this knowledge gap by a strong collaboration involving multidisciplinary approaches that include structural biochemistry of pre-mRNA splicing factors, molecular biology, and virology. Specifically, we will determine Tat- SF1 interactions among the human U2 snRNA and U2 snRNP subunits, and test these interfaces for functions in human splicing versus HIV-1 splicing and infectivity. We also will investigate a new mechanism of action for HIV-1 to pre-empt the human splicing machinery, in which the early stage HIV-1 Rev protein promotes late stage unspliced HIV-1 RNAs by disrupting the host Tat-SF1-U2 snRNP complex. Already we have: (i) produced diffracting crystals of the Tat-SF1 RNA recognition motif, (ii) obtained an initial structure of a complex between the Tat-SF1 and the SF3b155 subunit of the U2 snRNP, and (iii) generated a stable knockdown of Tat-SF1 in a human cell line. Moreover, our preliminary results demonstrate that the HIV- 1 Rev protein associates with human Tat-SF1. In addition to elucidating new mechanisms of action for HIV-1 to manipulate host pathways of gene expression, successful completion of the proposed experiments will set the stage for new therapeutic strategies that target a host cofactor, thereby circumventing drug resistance challenges that plague existing anti-HIV-1 treatments.