AMINOACYL TRNA SYNTHETASE SUBTRATE RECOGNITION

我的目标是建立一个充满活力的，独立的研究计划， 专注于研究RNA-蛋白质相互作用和结构- 生物系统中的功能关系。的主要功能 转移RNA（tRNA）分子是作为一个连接的网站， 氨基酸，然后被激活用于蛋白质合成。的 tRNA和氨酰-tRNA合成酶之间高度特异性， 催化这种附着的酶， 研究蛋白质-RNA原理的绝佳目标 互动转移RNA分子也可用作反向杂交的引物。 转录酶。这些酶在逆转录病毒中起作用， 从逆转录病毒RNA基因组转录DNA。 未来的发展方向 我的实验室将致力于了解艾滋病毒逆转 转录酶和人tRNA（Lys），这种特异性RNA引物， 病毒酶 简而言之，本研究的主要目标是解决 脯氨酸tRNA合成酶（ProRS）如何识别和相互作用的问题 与它的同源tRNA底物。我们已经取得了进展， 鉴定标记tRNA分子的核苷酸子集， 与脯氨酸的特异性氨酰化。我们还证明， 功能性tRNA（Pro）分子可以通过将两个 代表3 '-和5'-片段的寡核苷酸。使用这些 “退火的”底物，用化学合成的片段制备， 有助于识别RNA的结构特征 对ProRS的氨酰化很重要。的新用途 化学RNA合成将使共价探针的附着在 退火的tRNA底物的特异性内部位置， 交联和荧光研究。蛋白质结构 基序和特定的氨基酸残基与特定的 将鉴定tRNA（Pro）的区域。

My goal is to establish a vigorous, independent research program that focuses on the investigation of RNA-protein interactions and structure- function relationships in biological systems. The major function of transfer RNA (tRNA) molecules is to serve as a site of attachment for amino acids, which are then activated for use in protein synthesis. The high degree of specificity between tRNAs and aminoacyl-tRNA synthetases, the enzymes that catalyze this attachment, make these molecules excellent targets for studying the principles of protein-RNA interaction. Transfer RNA molecules are also used as primers by reverse transcriptases. These are enzymes in retroviruses that function to transcribe DNA from the retroviral RNA genome. A future direction of my lab will be to understand the interaction between HIV reverse transcriptase and human tRNA(Lys), the specific RNA primer for this viral enzyme. Briefly, the primary objective of this research is to address the question of how proline tRNA synthetase (ProRS) recognizes and interacts with its cognate tRNA substrate. We have already made progress in identifying the subset of nucleotides that mark a tRNA molecule for specific aminoacylation with proline. We have also demonstrated that a functional tRNA(Pro) molecule can be assembled by annealing together two oligonucleotides representing 3'- and 5'-fragments. The use of these "annealed" substrates, prepared with a chemically-synthesized fragment, has facilitated the identification of RNA structural characteristics that are important for aminoacylation by ProRS. Novel applications of chemical RNA synthesis will enable the attachment of covalent probes at specific, internal positions of annealed tRNA substrates for use in cross-linking and fluorescence studies. In this way, protein structural motifs and specific amino acid residues in close contact with specific regions of tRNA(Pro) will be identified.