Although the m.

3243A>G mutation was identified nearly three decades ago, the molecular consequences of the
mutation are still incompletely understood. Muscle cells with the m.3243A>G mutation grown in tissue culture
demonstrate respiratory deficiency [40]. King and Attardi developed a cell line called rho-0 that replicates in the absence
of mtDNA [41]. The rho-0 cells can be fused with cytoplasts harboring mutant mtDNA to form cybrids. Cybrids with
greater than 95% m.3243A>G mtDNA showed decreased rates of protein synthesis, lower levels of steady-state
mitochondrial translational products, reduced oxygen consumption, and increased amounts of an unprocessed RNA
fragment called RNA-19, which contained the mutant gene [42]. Other investigators have demonstrated that high levels
of the mutant tRNA decreased aminoacylation (covalent attachment of leucine to the tRNA) and were associated with
hypomodification of the D-stem. These alterations may contribute to decreased protein synthesis [43–45]. An alternative
theory also based on cybrid work is that the mutant tRNALeu