CAREER: Elucidating the Targets and Functions of Mammalian tRNA methyltransferase orthologs

本研究的目的是了解核糖核酸（RNA）分子如何影响细胞生长、发育和对环境的反应。目前的证据表明，RNA的功能可以通过特定酶添加到RNA上的化学基团（修饰）的存在来调节。然而，关于酶如何找到它们的目标RNA，修饰如何改变RNA的功能，以及修饰的RNA如何调节关键的细胞过程，还有很多有待研究。这项研究将解决这些问题，同时提供一个动态的研究环境，以促进科学领域代表性不足的群体（如少数民族和妇女）的科学培训和发展。为了分享研究期间开发的新技术方法，学生们将制作基于视频的教程，作为科学界的教学和培训资源。该项目还将为缺乏教育的高中生提供教育讲习班，以鼓励他们对生物学领域的好奇心和实验精神。据预测，哺乳动物基因组编码多种转移RNA甲基转移酶（Trm）酶，但大多数Trm蛋白的细胞功能尚不清楚。以下项目旨在通过：1)建立人类Trm酶的蛋白质相互作用网络，2)鉴定Trm蛋白的细胞靶点，以及3)确定Trm蛋白调节的细胞过程来阐明许多未表征的哺乳动物Trm酶的生物学作用。这些目标将通过综合生化、分子和遗传方法来实现，包括对Trm复合物的蛋白质组学分析，以及对具有改变的Trm蛋白的人类细胞的全局转录和翻译分析。通过这些发现，这项工作有望为新的理解奠定基础，了解由不同Trm酶催化的RNA修饰如何转化为帮助细胞对各种环境条件作出反应的生理输出。

The goal of this research is to understand how ribonucleic acid (RNA) molecules influence cell growth, development, and response to the environment. Current evidence shows that RNA function can be regulated by the presence of chemical groups (modifications) that are added to the RNA by specific enzymes. However, much remains to be learned about how the enzymes find their target RNAs, how the modifications change RNA functions, and how the modified RNAs regulate critical cellular processes. This research will address those questions, while providing a dynamic research environment to foster the scientific training and development of underrepresented groups in science such as minorities and women. To share new technical approaches developed during the research, students will produce video-based tutorials to serve as a teaching and training resource for the scientific community. The project will also provide educational workshops for underserved high school students to encourage curiosity and experimentation in the field of biology. Mammalian genomes are predicted to encode a diverse number of transfer RNA methyltransferase (Trm) enzymes, but the cellular functions for the majority of Trm proteins remain unknown. The following project aims to elucidate the biological roles of the numerous uncharacterized mammalian Trm enzymes by: 1) establishing the protein interaction networks of human Trm enzymes, 2) identifying the cellular targets of Trm proteins, and 3) determining the cellular processes modulated by Trm proteins. The goals will be achieved through an integrated biochemical, molecular and genetic approach involving proteomic analysis of Trm complexes together with global transcription and translation profiling of human cells with altered Trm proteins. Through these discoveries, the work promises to lay a foundation of new understanding of how the RNA modifications catalyzed by diverse Trm enzymes translate into physiological outputs that help cells respond to various environmental conditions.