Kinase Regulation of Nuclear Speckle Function and Splicing during Influenza Virus Infection

流感病毒感染期间核斑点功能和剪接的激酶调节

基本信息

批准号：
10685340
负责人：
MELANIE H. COBB
金额：
$ 56.51万
依托单位：
UNIVERSITY OF PENNSYLVANIA
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：
2016-05-25 至 2026-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10685340
关键词：
Alternative Splicing Biochemical Biochemistry Biological Assay Cell Nucleus Cells Chemicals Complement Creativeness Data Economic Burden Event Gene Expression Genes Genetic Transcription Goals Grant Health Human Immune response Impairment Infection Influenza Virus Infected Cells Integration Host Factors Label Link Maintenance Mass Spectrum Analysis Mediating Messenger RNA Methods Microscopy Morphology Nuclear Nuclear Proteins Nuclear RNA Phosphorylation Phosphotransferases Positioning Attribute Process Protein Kinase Proteins RNA RNA Processing RNA Splicing RNA-Binding Proteins Regulation Role Shapes Transcript Viral Viral Genes Viral Proteins Virus Virus Diseases Virus Replication Western Blotting Work experience genetic manipulation high resolution imaging influenza infection influenzavirus insight knock-down mRNA Precursor therapeutic target trafficking transcriptome sequencing viral RNA

项目摘要

PROJECT SUMMARY The goals of this proposal are to uncover how pre-mRNA splicing and nuclear speckles are controlled by the kinase TAO2, and to determine why TAO2 is required for successful Influenza virus (IAV) replication. Recent work has identified the understudied protein kinase TAO2 as a host factor essential for splicing and speckle localization of the IAV M RNA. A pool of TAO2 localizes to nuclear speckles and its loss, by chemical inhibition or protein depletion, alters nuclear speckle composition, splicing and export of IAV M RNA, and therefore impairs IAV replication. Inhibition of TAO2 also disrupts splicing of a subset of host mRNAs, without altering bulk host mRNA. These data uncover a new cellular activity for TAO2 and identify inhibition of TAO2 as a potential approach for controlling IAV infection. Preliminary data suggest that TAO2 interacts with several splicing factors and other RNA binding proteins. The work outlined in this proposal seeks a comprehensive understanding of the nuclear activities of TAO2 in human cells and the functional consequence of these activities for host and viral RNA expression. Specifically, a three-pronged approach will be taken to: (1) characterize functional TAO2 interaction partners and substrates of phosphorylation amongst nuclear proteins and determine if IAV infection alters these interactions or if TAO2 interacts directly with any IAV-encoded proteins; (2) define the role of TAO2 in maintaining the integrity of nuclear speckles and (3) characterize the global impact of TAO2 on the human splicing machinery and the consequence of this function for alternative splicing. These goals will be achieved through a combination of genetic manipulation of cells, biochemistry, mass spectrometry and microscopy. Importantly, the conclusions obtained from these studies will have implications for general mechanisms of RNA splicing and nuclear speckle formation and function and will further inform the understanding of host requirements and vulnerabilities for influenza infection.

项目概要该提案的目标是揭示前 mRNA 剪接和核斑点是如何被控制的激酶 TAO2，并确定为什么流感病毒 (IAV) 成功复制需要 TAO2。最近的工作已经确定了正在研究的蛋白激酶 TAO2 作为剪接和转录过程所必需的宿主因子。 IAV M RNA 的斑点定位。 TAO2 池定位于核斑点及其损失，通过化学抑制或蛋白质消耗，改变 IAV M 的核斑点组成、剪接和输出 RNA，因此会损害 IAV 复制。 TAO2 的抑制也会破坏宿主子集的剪接 mRNA，不改变大量宿主 mRNA。这些数据揭示了 TAO2 的新细胞活性并确定抑制TAO2作为控制IAV感染的潜在方法。初步数据表明 TAO2 与多种剪接因子和其他 RNA 结合蛋白相互作用。本提案中概述的工作寻求全面了解 TAO2 在人类细胞中的核活动及其功能这些活动对宿主和病毒 RNA 表达的影响。具体来说，要从三方面入手将用于：(1) 表征功能性 TAO2 相互作用伙伴和磷酸化底物并确定 IAV 感染是否改变这些相互作用或 TAO2 是否相互作用直接与任何 IAV 编码的蛋白质结合； (2)明确TAO2在维持核完整性中的作用 (3) 表征 TAO2 对人体剪接机制和该功能对于选择性剪接的结果。这些目标将通过组合来实现细胞遗传操作、生物化学、质谱和显微镜学。重要的是，从这些研究中获得的结论将对 RNA 剪接的一般机制产生影响和核散斑的形成和功能，并将进一步了解宿主的需求和流感感染的脆弱性。

项目成果

期刊论文数量（5）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Nucleoporin TPR is an integral component of the TREX-2 mRNA export pathway.

核孔蛋白 TPR 是 TREX-2 mRNA 输出途径的一个组成部分。

DOI：
发表时间：
2020-09-11
期刊：
影响因子：
16.6
作者：
Aksenova, Vasilisa;Smith, Alexandra;Lee, Hangnoh;Bhat, Prasanna;Esnault, Caroline;Chen, Shane;Iben, James;Kaufhold, Ross;Yau, Ka Chun;Echeverria, Carlos;Fontoura, Beatriz;Arnaoutov, Alexei;Dasso, Mary
通讯作者：
Dasso, Mary

Cryptic MHC-E epitope from influenza elicits a potent cytolytic T cell response.

流感病毒的隐性 MHC-E 表位可引发强效的溶细胞 T 细胞反应。

DOI：
发表时间：
2023-11
期刊：
影响因子：
30.5
作者：
Hogan, Michael J;Maheshwari, Nikita;Begg, Bridget E;Nicastri, Annalisa;Hedgepeth, Emma J;Muramatsu, Hiromi;Pardi, Norbert;Miller, Michael A;Reilly, Shanelle P;Brossay, Laurent;Lynch, Kristen W;Ternette, Nicola;Eisenlohr, Laurence C
通讯作者：
Eisenlohr, Laurence C

Aryl Sulfonamide Inhibits Entry and Replication of Diverse Influenza Viruses via the Hemagglutinin Protein.

芳基磺酰胺通过血凝素蛋白抑制多种流感病毒的进入和复制。

DOI：
发表时间：
2021-08-12
期刊：
影响因子：
7.3
作者：
White, Kris;Esparza, Matthew;Liang, Jue;Bhat, Prasanna;Naidoo, Jacinth;McGovern, Briana L;Williams, Michael A P;Alabi, Busola R;Shay, Jerry;Niederstrasser, Hanspeter;Posner, Bruce;García;Ready, Joseph;Fontoura, Beatriz M A
通讯作者：
Fontoura, Beatriz M A

Microglia at the scene of the crime: what their transcriptomics reveal about brain health.

犯罪现场的小胶质细胞：它们的转录组学揭示了大脑健康状况。