PetCR46, a gene which is essential for respiration and integrity of the mitochondrial genome

• Published in: Yeast (Chichester, England) Vol. 12; no. 6; pp. 577 - 582
• Main Authors: Coppée, J.‐Y, Rieger, K.‐J, Kaniak, A, Di Rago, J.‐P, Groudinsky, O, Slonimski, P.P
• Format: Journal Article
• Language: English
• Published: Chichester, UK John Wiley & Sons, Ltd 01.05.1996
• Subjects: Base Sequence; Blotting, Southern; Chromosome III; DNA, Mitochondrial - genetics; Genome; Meiosis; mitochondrial DNA; Molecular Sequence Data; Mutagenesis, Site-Directed; nuclear petite; Polymerase Chain Reaction; Reading Frames - genetics; Saccharomyces cerevisiae; Saccharomyces cerevisiae - cytology; Saccharomyces cerevisiae - genetics; Sequence Deletion; YCR46C
• ISSN: 0749-503X; 1097-0061
• DOI: 10.1002/(SICI)1097-0061(199605)12:6<577::AID-YEA950>3.0.CO;2-2

In the frame of the European Pilot Project for the functional analysis of newly discovered open reading frames (ORFs) from Saccharomyces cerevisiae chromosome III, we have deleted entirely the YCR46C ORF by a one‐step polymerase chain reaction method and replaced it by the HIS3 marker in the strain W303. The deletion has been checked by meiotic segregation and Southern blot analyses. Characterization of the deleted strain indicates that YCR46C is essential for respiration and maintenance of the mitochondrial genome since its deletion leads to the appearance of 100% of cytoplasmic petites. Hybridization with molecular probes from mtDNA of individual clones of such petites showed that about 50% did hybridize (rho⁻ clones) while others did not (possibly rho° clones). The wild‐type gene has been cloned and shown to complement the deletion. The gene, which probably codes for a mitochondrial ribosomal protein, has been called petCR46.