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Cellular Factors Critical for Initiation of HIV-1 Reverse Transcriptase

对 HIV-1 逆转录酶启动至关重要的细胞因素

基本信息

批准号：
10393691
负责人：
Karin M Musier-Forsyth
金额：
$ 39.84万
依托单位：
OHIO STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-09-01 至 2025-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10393691
关键词：
5&apos Untranslated Regions Acquired Immunodeficiency Syndrome Address Affect Affinity Amino Acyl Transfer RNA Antibodies Antiviral Agents Back Binding Binding Sites CRISPR/Cas technology Cell Nucleus Cell membrane Cells Co-Immunoprecipitations Complex Confocal Microscopy Cryoelectron Microscopy Development Dimerization Elements Event Fluorescence Resonance Energy Transfer Genetic Transcription Grant HIV-1 Human In Vitro Infection Lead Ligase Location Lysine-Specific tRNA Lysine-tRNA Ligase Measures Molecular Conformation Mutation Northern Blotting Nuclear Nuclear Export Nuclear Localization Signal Nucleic Acid Binding Pathway interactions Phosphorylation Process Protein Isoforms RNA RNA Conformation RNA-Directed DNA Polymerase Reporting Resolution Reverse Transcription Ribosomes Roentgen Rays Role Signal Transduction Specificity Structure Testing Time Transfer RNA Viral Virion Work base combat crosslink design genomic RNA insight new therapeutic target novel novel therapeutic intervention particle recruit scaffold single molecule therapeutic target transcriptome sequencing

项目摘要

Project Summary The primer for reverse transcription in HIV-1, human tRNALys3, is selectively packaged into virions along with tRNALys1,2. Human lysyl-tRNA synthetase (LysRS), the only cellular factor that interacts specifically with both tRNALys isoacceptors, is also packaged into HIV-1. Selective packaging of tRNALys depends on the ability of LysRS to bind to tRNALys and the presence of both host cell factors is required for optimal viral infectivity. LysRS is normally part of a dynamic mammalian multi-synthetase complex. In the previous grant period, we made substantial progress towards elucidating the mechanism by which human LysRS is recruited into HIV-1 particles. We reported that HIV-1 infection results in a free pool of phosphorylated LysRS (pS207-LysRS) that is partially re- localized to the nucleus of target cells. Blocking this pathway in HIV-1 producing cells abolished LysRS packaging and resulted in less infectious progeny virions. We also extended earlier work showing that LysRS binds to a tRNA-like element in the primer-binding site (PBS) region of HIV-1 genomic RNA (gRNA), which facilitates tRNA release from LysRS and primer placement onto the PBS. In the next grant period, we will build on these results by examining the overall hypotheses that phosphorylated LysRS facilitates uncharged tRNA primer packaging and that tRNALys3 annealing to the PBS alters gRNA conformation and dynamics, thereby impacting downstream events that are critical for the HIV-1 lifecycle. This work will transform our understanding of the role of two host cell factors in HIV-1 replication. Aim 1 will explore the role of LysRS phosphorylation in packaging an uncharged form of tRNALys3 and validate this isoform of LysRS as a therapeutic target. Aim 2 will establish the effect of tRNA annealing and pS207-LysRS binding on gRNA structure and dynamics and provide new insights into the timing of this key step in the viral lifecyle, which may reveal a novel therapeutic strategy.

项目摘要 HIV-1逆转录的引物（人tRNALys 3）与一起沿着地选择性包装到病毒粒子中 tRNALys1，2.人赖氨酰-tRNA合成酶（LysRS），唯一与两者特异性相互作用的细胞因子 tRNALys同种受体也被包装到HIV-1中。tRNALys的选择性包装取决于最佳病毒感染性需要LysRS与tRNALys结合以及两种宿主细胞因子的存在。LysRS 通常是动态哺乳动物多合成酶复合体的一部分。在过去的一段时间里，我们在阐明人LysRS被募集到HIV-1颗粒中的机制方面取得了实质性进展。我们报道了HIV-1感染导致磷酸化LysRS（pS207-LysRS）的游离池，其部分地被重新激活。定位于靶细胞的细胞核。在HIV-1产生细胞中阻断该途径可消除LysRS 包装并导致感染性较低的子代病毒体。我们还扩展了早期的工作，表明LysRS 与HIV-1基因组RNA（gRNA）的引物结合位点（PBS）区域中的tRNA样元件结合，促进tRNA从LysRS释放并将引物放置到PBS上。在下一个资助期内，我们将通过检查磷酸化LysRS促进不带电荷的tRNA的总体假设，引物包装和与PBS退火的tRNALys 3改变了gRNA构象和动力学，从而影响对HIV-1生命周期至关重要的下游事件。这项工作将改变我们对两种宿主细胞因子在HIV-1复制中的作用。目的1将探讨LysRS磷酸化在包装不带电荷的tRNALys 3形式，并验证LysRS的这种亚型作为治疗靶点。目标2将建立tRNA退火和pS207-LysRS结合对gRNA结构和动力学的影响，并提供对病毒生命周期中这一关键步骤的时机有了新的认识，这可能揭示一种新的治疗策略。

项目成果

期刊论文数量（9）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Anticodon-like binding of the HIV-1 tRNA-like element to human lysyl-tRNA synthetase.

DOI：
10.1261/rna.058081.116
发表时间：
2016-12
期刊：
RNA (New York, N.Y.)
影响因子：
0
作者：
Liu S;Comandur R;Jones CP;Tsang P;Musier-Forsyth K
通讯作者：
Musier-Forsyth K

Human lysyl-tRNA synthetase phosphorylation promotes HIV-1 proviral DNA transcription.

DOI：
10.1093/nar/gkad941
发表时间：
2023-12-11
期刊：
NUCLEIC ACIDS RESEARCH
影响因子：
14.9
作者：
Tang, Yingke;Behrens, Ryan T.;St Gelais, Corine;Wu, Siqi;Vivekanandan, Saravanan;Razin, Ehud;Fang, Pengfei;Wu, Li;Sherer, Nathan;Musier-Forsyth, Karin
通讯作者：
Musier-Forsyth, Karin

Hairpin RNA-induced conformational change of a eukaryotic-specific lysyl-tRNA synthetase extension and role of adjacent anticodon-binding domain.

发夹 RNA 诱导真核特异性赖氨酰-tRNA 合成酶延伸的构象变化以及相邻反密码子结合域的作用。

DOI：
10.1074/jbc.ra120.013852
发表时间：
2020
期刊：
The Journal of biological chemistry
影响因子：
0
作者：
Liu,Sheng;Refaei,Maryanne;Liu,Shuohui;Decker,Aaron;Hinerman,JenniferM;Herr,AndrewB;Howell,Mike;Musier-Forsyth,Karin;Tsang,Pearl
通讯作者：
Tsang,Pearl

Phosphomimetic S207D Lysyl-tRNA Synthetase Binds HIV-1 5'UTR in an Open Conformation and Increases RNA Dynamics.