Human Mitochondrial tRNA Structure and Function

描述（由申请人提供）：越来越多的人类疾病与线粒体基因组内转移RNA（tRNA）基因的点突变相关。与人类线粒体（hs mt）tRNA突变相关的疾病的因果基础是医学和细胞生物学研究中的一个积极研究领域，但有关这些tRNA性质的信息有限，阻碍了这些线粒体病理学的分子基础的描绘。 我们的目标是描述人类线粒体中独特的tRNA功能，并阐明与疾病相关的hs mt tRNA突变体的功能和结构特性是如何受到干扰的。将通过以下方法研究hs mt tRNA中致病性突变的影响：1）使用溶液足迹法、体外选择、计算和光谱方法监测结构性质，2）使用多种体外测定研究功能性质，包括氨酰化和核糖体蛋白质合成的效率和保真度，和3）使用人细胞系和细菌模型系统在体内探测这些分子的功能。这种多管齐下的方法将允许为这类未表征的分子制定结构/功能关系，并将提供有关疾病相关tRNA突变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.