Characterization of a Non-Spliceosomal mRNA Ligase

描述（申请人提供）：mRNA反式连接现象是一个知之甚少的过程，需要新的实验技术和见解。本提案将研究人类癌细胞中mRNA反式结扎，该细胞被发现是对临床相关核糖核酸酶治疗的反应。核糖核酸酶诱导的mRNA反式连接的程度将通过RNA测序技术来表征。假设负责观察到的mRNA连接的酶将被动力学和机械地表征。结合细胞研究和mRNA连接的体外表征将揭示这一未表征的生理过程的重要细节。对mRNA反式连接的基本理解可能导致对各种正常生理和疾病过程的理解。

DESCRIPTION (provided by applicant): The phenomenon of mRNA trans-ligation is a poorly understood process in need of novel experimental techniques and insight. This proposal will study mRNA trans-ligation in human cancer cells that was discovered as a response to treatment with a clinically relevant ribonuclease. The extent of mRNA trans- ligation induced by ribonucleases will be characterized by RNA sequencing techniques. An enzyme hypothesized to be responsible for the observed mRNA ligation will be characterized kinetically and mechanistically. A combination of cellular studies and in vitro characterizations of mRNA ligation will bring to light important details about this uncharacterized physiological process. A fundamental understanding of mRNA trans-ligation is likely to lead to an understanding of various normal physiological and disease processes. PUBLIC HEALTH RELEVANCE: The proposed research will investigate the uncharacterized phenomenon of mRNA trans-ligation. A fundamental understanding of this process will be important to understand the role mRNA trans-ligation plays in both normal physiological and various disease processes. Characterization of the enzyme involved in this process will facilitate the design of inhibitors that specifically target mRNA trans-ligation.