Localization of RNA binding proteins in the control of cell survival

RNA结合蛋白在细胞存活控制中的定位。(5000字符最大值） 我实验室的研究项目集中在了解不同的RNA结合蛋白如何调节细胞应激反应。在过去的10年里，我们把注意力集中在这样一个蛋白，hnRNP A1，它在RNA代谢中发挥着不同的作用，包括端粒修复，mRNA剪接，mRNA输出，应激颗粒形成和选择性mRNA翻译的病毒和细胞的mRNA。我们已经表明，在响应不同的细胞应激，如紫外线照射，渗透压休克或病毒感染，hnRNP A1积累在受影响的细胞的细胞质中，并调节mRNA的稳定性或选择性翻译的mRNA编码的关键凋亡调节蛋白cIAP 1，XIAP，Bcl-xL和Apaf-1。此外，hnRNP A1的细胞质积累是肠道病毒（如脊髓灰质炎病毒、肠道病毒和鼻病毒，所有重要的人类病原体）生命周期进展中的关键步骤。然而令人惊讶的是，哪些信号通路调节hnRNP A1的细胞质积累，以及这种调节是如何发挥的是非常不清楚的。 最近，siRNA技术和自动化高通量成像技术的进步允许对复杂生物网络进行系统的询问。我们已经应用这些技术来研究亚细胞定位的hnRNP A1在渗透胁迫或人鼻病毒感染。我们的初步数据确定了几个新的激酶，调节hnRNPA 1在细胞中的定位。我们的工作假设是，不同的激酶和信号通路控制亚细胞定位的hnRNP A1在定义的生理和病理刺激。该计划的长期目标是详细了解细胞信号通路如何与离散RNA顺式调节元件交叉，以在不同的生理条件下选择性地调节基因表达。为了实现这一愿景，本提案的具体短期研究目标是： 1.目的：明确hnRNPA 1激酶如何调控hnRNPA 1的定位。 2.探讨调控这些信号通路对细胞存活和病毒生命周期的影响及其机制。 3.鉴定和表征调节hnRNP A1在病毒感染后细胞质蓄积的激酶。 虽然我们最初的重点是hnRNPA 1，我们将确定的激酶可能有其他目标，可能是重要的细胞应激反应。我们的长期计划是利用我们的资源和专业知识，并使用这些激酶来研究其他RBP及其在细胞存活和病毒感染中的作用。

Localization of RNA binding proteins in the control of cell survival. (5000 char maximum) The research program in my laboratory is focused on understanding how distinct RNA binding proteins regulate cellular stress response. Over the past 10 years we focused our attention on one such protein, hnRNP A1, which plays diverse roles in RNA metabolism, including telomere repair, mRNA splicing, mRNA export, stress granule formation and selective mRNA translation of viral and cellular mRNAs. We have shown that in response to diverse cellular stresses, such as UV irradiation, osmotic shock or viral infection, hnRNP A1 accumulates in the cytoplasm of affected cells and regulates either mRNA stability or selective translation of mRNAs encoding key apoptosis-regulating proteins cIAP1, XIAP, Bcl-xL and Apaf-1. In addition, cytoplasmic accumulation of hnRNP A1 is a critical step in the life cycle progression of Enteroviruses , such as poliovirus, enterovirus, and rhinovirus, all important human pathogens. Yet surprisingly, which signaling pathways regulate cytoplasmic accumulation of hnRNP A1, and how is this regulation exerted is very poorly understood. Recently, advances in siRNA technologies and automated high throughput imaging technologies allowed for systematic interrogations of complex biological networks. We have applied these technologies to study subcellular localization of hnRNP A1 in response to osmotic stress or human rhinovirus infection. Our preliminary data identified several novel kinases that modulate hnRNP A1 localization in cells. Our working hypothesis is that distinct kinases and signaling pathways control subcellular localization of hnRNP A1 in response to defined physiological and pathological stimuli. The long-term goal of the program is to gain a detailed understanding of how cellular signaling pathways intersect with discrete RNA cis-regulatory elements to selectively regulate gene expression under distinct physiological conditions. To achieve this vision the specific short term research objectives of this proposal are: 1. To define how hnRNPA1 kinases control hnRNP A1 localization. 2. To investigate the mechanism and biological consequences of modulating these signaling pathways on cell survival and viral life cycle. 3. To identify and characterize kinases that regulate cytoplasmic accumulation of hnRNP A1 in response to viral infection. Although our initial focus is on hnRNP A1, the kinases that we will identify may have other targets that could be important for cellular stress response. Our long term plan is to use our resources and expertise and use these kinases to study other RBPs and their role in cell survival and viral infection.