Mechanisms and Regulation of Yeast Internal Ribosome Entry Sites

描述（由申请人提供）：我的主要科学兴趣是基因表达的转录后调控。特别是，我着迷于RNA依赖的翻译控制机制的多样性和复杂性。将来，我希望把自己的实验室建成一个研究基因表达的基本机制和导致人类疾病的基因表达缺陷的杰出中心。在加州大学伯克利分校的Jennifer Doudna实验室继续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，以及多样化和刺激的智力环境。我还将发展专业技能，我将需要在未来作为一个独立的调查员茁壮成长，通过无数的机会来监督学生与不同的科学优势和兴趣。 翻译起始是真核生物基因表达调控的关键环节。响应于各种环境损伤，细胞下调翻译的主要（帽依赖性）机制，同时通过替代起始机制（包括将翻译机制直接募集到内部核糖体进入位点（IRES））来促进选择的一组基因的表达。关于细胞IRES活性或这些机制的信息特异性调节的分子基础知之甚少。我已经鉴定了24个在体外和体内都具有强活性的酵母细胞IRES元件，为联合收割机结合分子遗传学和生物化学以确定IRES依赖性基因表达的分子基础和生理意义创造了独特的机会。我的目的是确定细胞IRES RNA序列及其相关RNA结合蛋白的分子功能，并最终，利用生物化学、分子遗传学和生物信息学方法的组合，确定将应激相关信号传导途径与全局和信息特异性翻译调节联系起来的机制。从这项工作的结果将提供详细的分子洞察的机制和内部启动的翻译不同的细胞IRES，这是预期有广泛的影响，真核基因表达的翻译控制的调控。

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.