Yeast mitochondrial RNase P: sequence of the RPM2 gene and demonstration that its product is a protein subunit of the enzyme

• Published in: The Journal of biological chemistry 1993-09, Vol.268 (26), p.19791-19796
• Main Authors: Dang, Y L, Martin, N C
• Format: Article
• Language: English
• Subjects: actividad enzimatica; activite enzymatique; Amino Acid Sequence; Analytical, structural and metabolic biochemistry; Base Sequence; Biological and medical sciences; Cloning, Molecular; Electrophoresis, Polyacrylamide Gel; Endoribonucleases - biosynthesis; Endoribonucleases - genetics; Endoribonucleases - isolation & purification; Enzymes and enzyme inhibitors; enzymic activity; Fundamental and applied biological sciences. Psychology; gene; genes; Genes, Fungal; Hydrolases; Macromolecular Substances; Mitochondria - enzymology; Molecular Sequence Data; Molecular Weight; nucleotide sequence; proteinas; proteine; proteins; Recombinant Fusion Proteins - biosynthesis; Recombinant Fusion Proteins - isolation & purification; Restriction Mapping; ribonucleasas; ribonuclease; Ribonuclease P; ribonucleases; RNA, Catalytic - biosynthesis; RNA, Catalytic - genetics; RNA, Catalytic - isolation & purification; RNA, Fungal - genetics; RNA, Fungal - isolation & purification; Saccharomyces cerevisiae; Saccharomyces cerevisiae - enzymology; Saccharomyces cerevisiae - genetics; secuencia nucleica; sequence nucleique
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1016/S0021-9258(19)36583-4

We report here the sequence of the RPM2 gene which codes for the 105-kDa protein previously purified from the mitochondria of Saccharomyces cerevisiae and shown by genetic techniques to be required for mitochondrial RNase P activity. The sequence predicts a primary translation product of 1202 residues with a molecular mass of 139 kDa and no obvious sequence similarity to any known protein in the data bases. There are 122 amino-terminal amino acids predicted by the gene that are not found in the purified protein, some of which may play a role in mitochondrial targeting of the protein. Antibodies raised against a trpE- 105-kDa fusion protein recognize a 105-kDa protein in wild-type cells but not in cells carrying a disruption of the RMP2 gene. Immune, but not preimmune serum, immunoprecipitates the RNase P RNA and the mitochondrial RNase P activity. Thus, the 105-kDa protein forms a complex with RNase P RNA and is required for RNase P activity as predicted for a bona fide subunit of the enzyme.