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Mechanisms regulating poxvirus post-replicative protein synthesis

痘病毒复制后蛋白质合成的调节机制

基本信息

批准号：
10666342
负责人：
Zhilong Yang
金额：
$ 37.88万
依托单位：
TEXAS A&M AGRILIFE RESEARCH
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-07-01 至 2025-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10666342
关键词：
7-methylguanosine Animals Antineoplastic Agents Area Automobile Driving Cells DNA biosynthesis Development Gene Expression Gene Expression Regulation Genes Human Individual Knowledge Mediating Messenger RNA Molecular Morbidity - disease rate Oncolytic Outcome Peptide Initiation Factors Plasmids Poly(A)-Binding Proteins Poxviridae Protein Biosynthesis Proteins Ribosomal Proteins Role Site Smallpox Testing Transcript Translating Translation Initiation Translational Repression Translations Vaccines Vaccinia virus Viral Virion Virus Virus Diseases Virus Replication biothreat cancer therapy decapping enzyme experimental study expression vector improved mRNA Decay mRNA Transcript Degradation mRNA Translation mortality mutant poxvirus vectors protein expression prototype recruit translation factor vaccine development vaccinia virus vector vector vaccine viral DNA

项目摘要

Project Summary Although smallpox had been eradicated, the chance the virus could arise from unknown stocks or de novo synthesis remains a credible biothreat. Moreover, other poxviruses cause significant morbidity and mortality in humans and economically important animals; and poxviruses are widely being developed as vaccine and protein expression vectors, and anti-cancer agents. Poxvirus gene expression is an arguably irreplaceable area in elucidating fundamental mechanisms of poxvirus replication and developing poxvirus-based utilities. Vaccinia virus (VACV) is the prototype poxvirus. VACV post-replicative genes (those expressed after viral DNA replication) number over 90 and encode mainly virion-associated proteins. They are more efficiently translated than cellular mRNAs and expressed at high levels. The objective of this project is to elucidate molecular mechanisms driving selective translation of VACV post-replicative mRNAs, all of which have an unusual feature—a 5'-poly(A) leader. During the post-replicative stage of VACV infection, the poly(A) leader appears to confer a translational advantage. The central hypothesis of this project is that the 5'-poly(A) leader coordinates with specific viral and cellular factors to promote translation of VACV post-replicative mRNAs. Supporting this hypothesis, D9 and D10, two VACV-encoded decapping enzymes that are known to promote cellular and viral mRNA degradation, are required for efficient translation of VACV post-replicative mRNAs. Exogenous expression of D10 in uninfected cells promotes translation of mRNA with a 5'-poly(A) leader. This is striking given that activation of cellular decapping enzymes causes translation repression. Moreover, La-related protein 4 (LARP4), a cellular poly (A)-binding protein and mRNA translation regulator, is required for VACV post- replicative protein synthesis. LARP4 is also highly enriched in viral factories, where viral post-replicative mRNA translation occurs. Three Specific Aims will be carried out to test how these factors promote selectively translation of VACV post-replicative mRNAs. In Aim 1, the role and molecular determinants of poxvirus decapping enzymes in promoting viral post-replicative mRNA translation will be defined. In Aim 2, the mechanisms by which VACV decapping enzymes promote viral post-replicative mRNA translation will be elucidated. In Aim 3, how LARP4 regulates VACV post-replicative mRNA translation will be determined. The results will provide an integrated new paradigm illustrating how an intrinsic sequence in viral transcripts, specific viral and cellular factors orchestrate selective translation of VACV post-replicative mRNAs, and in turn, VACV replication. The information can be used to tune or block poxvirus replication and improve the efficacy of VACV vectors used in vaccine and oncolytic cancer therapy. Moreover, these findings can provide knowledge for understanding fundamental mechanisms of eukaryotic translation.

项目摘要虽然天花已经被根除，但病毒可能来自未知的种群或新生的机会仍然很小。合成仍然是一种可信的生物威胁。此外，其他痘病毒在哺乳动物中引起显著的发病率和死亡率。人类和经济上重要的动物;并且痘病毒被广泛开发为疫苗，蛋白质表达载体和抗癌剂。痘病毒基因表达是一种无可替代的在阐明痘病毒复制的基本机制和开发基于痘病毒的实用程序领域。牛痘病毒（VACV）是痘病毒的原型。VACV复制后基因（在病毒DNA扩增后表达的基因）复制）数量超过90，主要编码病毒体相关蛋白。翻译效率更高比细胞mRNA高，并且表达水平高。本项目的目的是阐明驱动VACV复制后mRNA选择性翻译的机制，所有这些都具有不寻常的特征-5 '-聚（A）前导。在VACV感染的复制后阶段，poly（A）前导序列似乎赋予翻译优势。该项目的中心假设是，5 '-poly（A）前导协调与特定的病毒和细胞因子一起促进VACV复制后mRNA的翻译。支持这一假设，D9和D10，两种VACV编码的去帽酶，已知其促进细胞和病毒的生长， mRNA降解是VACV复制后mRNA有效翻译所必需的。外源 D10在未感染细胞中的表达促进具有5 ′-聚腺苷酸前导序列的mRNA的翻译。这是惊人的假定细胞去帽酶的激活引起翻译抑制。此外，La相关蛋白 4（LARP 4），一种细胞poly（A）结合蛋白和mRNA翻译调节因子，是VACV后复制性蛋白质合成LARP 4还高度富集在病毒工厂中，其中病毒复制后mRNA 翻译发生了。三个具体目标将进行测试，这些因素如何促进选择性 VACV复制后mRNA的翻译。目的1痘病毒的作用和分子决定因素将定义在促进病毒复制后mRNA翻译中的脱帽酶。在目标2中， VACV脱帽酶促进病毒复制后mRNA翻译的机制将被阐明。在目标3中，将确定LARP 4如何调节VACV复制后mRNA翻译。的结果将提供一个综合的新范例，说明病毒转录物中的内在序列，特异性病毒和细胞因子协调VACV复制后mRNA的选择性翻译，反过来， VACV复制。这些信息可用于调节或阻断痘病毒复制，并提高免疫治疗的疗效。 VACV载体用于疫苗和溶瘤癌症治疗。此外，这些发现可以提供知识，理解真核生物翻译的基本机制。