Human Mitochondrial tRNA Structure and Function

描述(申请人提供)：越来越多的人类疾病与线粒体基因组中转移RNA(TRNA)基因的点突变有关。人类线粒体(Hs Mt)tRNAs突变相关疾病的因果基础是医学和细胞生物学研究中一个活跃的研究领域，但关于这些tRNAs性质的信息有限阻碍了这些线粒体病理的分子基础的描绘。 我们的目标是确定人类线粒体中独特的tRNAs的功能，并阐明hs mt tRNA突变体与疾病相关的功能和结构特性是如何受到干扰的。Hs mt tRNA致病突变的影响将通过以下方式进行研究：1)使用溶液足迹法、体外选择、计算和光谱方法监测结构特性；2)使用各种体外分析方法研究功能特性，包括氨基酰化和核糖体蛋白质合成的效率和保真度；以及3)使用人类细胞系和细菌模型系统探索这些分子在体内的功能。这种多管齐下的方法将允许对这类未表征的分子的结构/功能关系进行公式化，并将提供与疾病相关的tRNA突变的影响的分子水平信息。

DESCRIPTION (provided by applicant): A growing number of human diseases are correlated with point mutations in transfer RNA (tRNA) genes within the mitochondrial genome. The cause-and-effect basis for diseases associated with mutations in human mitochondrial (hs mt) tRNAs is an area of active investigation in medical and cell biology research, but the limited availability of information concerning the properties of these tRNAs has hampered the delineation of the molecular basis of these mitochondrial pathologies. Our goal is to characterize the unique tRNAs functioning in human mitochondria, and to elucidate how the functional and structural properties of hs mt tRNA mutants associated with disease are perturbed. The effects of pathogenic mutations in hs mt tRNAs will be investigated by: 1) monitoring structural properties using solution footprinting, in vitro selection, computational, and spectroscopic methods, 2) investigating functional properties, including the efficiency and fidelity of aminoacylation and ribosomal protein synthesis, using a variety of in vitro assays, and 3) probing the function of these molecules in vivo using human cell lines and a bacterial model system. This multipronged approach will allow the formulation of structure/function relationships for this uncharacterized class of molecules and will provide molecular-level information concerning the effects of disease-related tRNA mutations.