Control of elF4F complex by Vesicular Stomatitis Virus

水泡性口炎病毒对 eF4F 复合物的控制

• 批准号: 6909396
• 负责人: John H Connor
• 金额: $ 16.2万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-08-01 至 2008-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6909396
• 关键词: Vesiculovirus; infection; proteins

One of the most striking features of gene expression in cells infected with many different viruses is that there is an inhibition of translation of host mRNAs while viral mRNAs are being translated. This viral control of translation plays a critical role in viral pathogenesis in intact animal hosts. This proposal is focused on understanding the mechanism by which the prototype negative strand RNA virus, vesicular stomatitis virus (VSV), interacts with the translation initiation factor complex eIF4F. eIF4F is a multi-protein complex, and its function is know to be the target of many viruses such as poliovirus, adenovirus and rotavirus. The aims of this project are to determine how host VSV infection causes the dephosphorylation of eIF4E, the cap-binding subunit of the eIF4F complex. This will be accomplished by directly analyzing the activity of the eIF4E and eIF4E binding protein (eIF4E-BP1) kinase and phosphatase activity in infected cell lysates, and by determining whether the eIF4E kinases are dissociated from eIF4F during VSV infection. Additionally, the aims of this proposal are to determine whether there are additional changes to the protein makeup of the eIF4F protein complex by using standard affinity isolation techniques followed by immunoblot analysis and/or mass-spec sequencing following complex purification. These experiments will help understand how VSV, a virus that produces 5' capped, and 3' polyadenylated mRNAs manages to dominate the host translation apparatus, excluding cellular capped and polyadenylated host mRNA and translating its own.

感染多种不同病毒的细胞中基因表达最显着的特征之一是，在翻译病毒 mRNA 时，宿主 mRNA 的翻译受到抑制。 这种病毒对翻译的控制在完整动物宿主的病毒发病机制中起着至关重要的作用。 该提案的重点是了解原型负链 RNA 病毒、水泡性口炎病毒 (VSV) 与翻译起始因子复合物 eIF4F 相互作用的机制。 eIF4F是一种多蛋白复合物，其功能已知是脊髓灰质炎病毒、腺病毒和轮状病毒等多种病毒的靶标。 该项目的目的是确定宿主 VSV 感染如何导致 eIF4E（eIF4F 复合物的帽结合亚基）去磷酸化。 这将通过直接分析受感染细胞裂解物中的 eIF4E 和 eIF4E 结合蛋白 (eIF4E-BP1) 激酶的活性和磷酸酶活性，并确定在 VSV 感染期间 eIF4E 激酶是否从 eIF4F 解离来完成。 此外，该提案的目的是通过使用标准亲和分离技术，然后在复合物纯化后进行免疫印迹分析和/或质谱测序，确定 eIF4F 蛋白复合物的蛋白质组成是否存在其他变化。 这些实验将有助于了解 VSV（一种产生 5' 加帽和 3' 聚腺苷酸化 mRNA 的病毒）如何设法控制宿主翻译装置，排除细胞加帽和聚腺苷酸化宿主 mRNA 并翻译自己的 mRNA。