41] Analysis of human mitochondrial RNA

Abstract

Publisher Summary This chapter focuses on the human mitochondrial RNA. The powerful molecular techniques have allowed the dissection of the mammalian, in particular human, mitochondrial genome with a resolution that has approached and often surpassed that of the finest genetic analysis. Two fundamental approaches are followed in these investigations. First, with the availability of the cloning technology and of rapid sequencing methods, it has been possible to determine the complete sequence of mitochondrial DNA (mtDNA) from several mammalian species and the partial sequence of mtDNA from other species. Second, the transcription of mtDNA in HeLa cells is analyzed in depth, and the structural, metabolic, and mapping properties of the transcription products are described. The availability of a method of HeLa cell growth giving a high yield of cells and the development of in vivo labeling procedures producing mitochondrial nucleic acids of high specific activity have alleviated the difficulties associated with the presence of the mitochondrial RNA species in extremely small amounts. Sensitive techniques are used for sequencing, mapping, and studying the metabolism of mitochondrial RNAs.