Mechanisms and Regulation of Yeast Internal Ribosome Entry Sites

描述(申请人提供)：我的主要科学兴趣是基因表达的转录后调控。特别是，我对依赖于RNA的翻译控制机制的多样性和复杂性很感兴趣。在未来，我希望把我自己的实验室打造成一个卓越的中心，研究基因表达的基本机制以及导致人类疾病的基因表达缺陷。在加州大学伯克利分校詹妮弗·杜德纳的实验室接受我的培训，将提供与翻译相关的RNA-蛋白质相互作用的生化解剖方面的出色技术教育，以及一个多样化和激动人心的智力环境。我还将通过无数机会监督具有不同科学优势和兴趣的学生，发展我未来作为一名独立调查员所需的专业技能。 翻译起始是真核基因表达调控的关键环节。作为对各种环境侮辱的反应，细胞下调初级(帽子依赖的)翻译机制，同时通过替代的启动机制，包括将翻译机制直接招募到内部核糖体进入位点(IRESS)，促进一组特定基因的表达。对细胞IRES活性的分子基础或对这些机制的信息特异性调控知之甚少。我已经在体外和体内鉴定了24个具有很强活性的酵母细胞IRES元件，创造了一个独特的机会，将分子遗传学和生物化学结合起来，以确定IRES依赖基因表达的分子基础和生理意义。我的目标是定义细胞IRES RNA序列及其相关RNA结合蛋白的分子功能，并最终利用生化、分子遗传学和生物信息学方法确定将胁迫相关信号通路与全球和信息特异性翻译调控联系起来的机制。这项工作的结果将为不同细胞IRESS内部启动翻译的机制和调控提供详细的分子洞察力，有望对真核基因表达的翻译调控具有广泛的意义。

DESCRIPTION (provided by applicant): My chief scientific interest is post-transcriptional regulation of gene expression. In particular, I am fascinated by the diversity and complexity of RNA-dependent mechanisms of translational control. In the future, I hope to make my own lab an outstanding center for research into the fundamental mechanisms of gene expression and the defects in gene expression that lead to human diseases. Pursuing my training in Jennifer Doudna's lab at UC Berkeley will provide an outstanding technical education in biochemical dissection of RNA-protein interactions relevant to translation, as well as a diverse and stimulating intellectual environment. I will also develop the professional skills I will need to thrive as an independent investigator in the future, through numerous opportunities to supervise students with diverse scientific strengths and interests. Translation initiation is a crucial point of regulation of eukaryotic gene expression. In response to a variety of environmental insults, cells down-regulate the primary (cap-dependent) mechanism of translation while favoring expression of a select group of genes via alternative initiation mechanisms including direct recruitment of the translation machinery to internal ribosome entry sites (IRESs). Little is known about the molecular basis for cellular IRES activity or for message-specific regulation of these mechanisms. I have identified 24 yeast cellular IRES elements with strong activity both in vitro and in vivo, creating a unique opportunity to combine molecular genetics and biochemistry to determine the molecular basis for and physiological significance of IRES-dependent gene expression.My aim is to define the molecular functions of cellular IRES RNA sequences and their associated RNA-binding proteins and ultimately, to identify the mechanisms linking stress-related signaling pathways to global and message-specific translational regulation using a combination of biochemical, molecular genetic, and bioinformatic approaches. Results from this work will provide detailed molecular insight into the mechanisms and regulation of internal initiation of translation by diverse cellular IRESs, which is expected to have wide-ranging implications for translational control of eukaryotic gene expression.