Mutational studies of the major tRNA region of the S. cerevisiae mitochondrial genome

The major tRNA genes in S. cerevisiae mitochondria are contained within a 20 kb segment of the mitochondrial DNA. In order to analyze the functional role of this region we have isolated several mitochondrial mutations, which are temperature-sensitive for growth on non-fermentable carbon sources. These mutations, localized in the major tRNA cluster region, can be classified in different groups according to their (a) genetic and physical localization, (b) spectrum of suppression and (c) biochemical characteristics. Some of these are mutations in tRNA genes which affect tRNA function; others alter the synthesis of the gene product. Finally, we found two mutations localized in, or in the vicinity of, the open reading frame RF2. RF2 has been postulated to be a maturase-like protein (Michel 1984) but no function for it has yet been demonstrated. The existence of defective mutants may confirm that RF2 is indeed necessary for mitochondrial biogenesis and so allow for a study of the expression of this gene.