Skip to main content
SpringerLink
Log in

Overexpression of a novel member of the mitochondrial carrier family rescues defects in both DNA and RNA metabolism in yeast mitochondria

  • Original Paper
  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Abstract

The PIF1 and MRS2 gene products have previously been shown to be essential for mitochondrial DNA maintenance at elevated temperatures and mitochondrial group II intron splicing, respectively, in the yeast Saccharomyces cerevisiae. A multicopy suppressor capable of rescuing the respiratory deficient phenotype associated with null alleles of either gene has been isolated. This suppressor is a nuclear gene that was called RIM2/MRS12. The RIM2/MRS12 gene encodes a predicted protein of 377 amino acids that is essential for mitochondrial DNA metabolism and proper cell growth. Inactivation of this gene causes the total loss of mitochondrial DNA and, compared to wild-type rhoo controls, a slow-growth phenotype on media containing glucose. Analysis of the RIM2/MRS12 protein sequence suggests that RIM2/MRS12 encodes a novel member of the mitochondrial carrier family. In particular, a typical triplicate structure, where each repeat consists of two putative transmembrane segments separated by a hydrophilic loop, can be deduced from amino acid sequence comparisons and the hydropathy profile of RIM2/MRS12. Antibodies directed against the aminoterminus of RIM2/MRS12 detect this protein in mitochondria. The function of the RIM2/MRS12 protein and the substrates it might transport are discussed.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Adrian GS, McCammon MT, Montgomery DL, Douglas MG (1986) Sequences required for delivery and localization of the ADP/ATP translocator to the mitochondrial inner membrane. Mol Cell Biol 6:626–634

    Google Scholar 

  • Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1990) Basic local alignment research tool. J Mol Biol 215:403–410

    Google Scholar 

  • Aquila H, Link TA, Klingenberg M (1987) Solute carriers involved in energy transfer in mitochondria form a homologous family. FEBS Lett 212:1–9

    Google Scholar 

  • Bairoch A (1993) The PROSITE dictionary of sites and patterns in proteins, its current status. Nucleic Acids Res 21:3097–3103

    Google Scholar 

  • Bonneaud N, Ozier-Kalogeropoulos O, Li G, Labouesse M, Minvielle-Sebastia L, Lacroute F (1991) A family of low and high copy replicative, integrative and single-stranded S. cerevisiae/E. coli shuttle vectors. Yeast 7:609–615

    Google Scholar 

  • Bouillaud F, Weissenbach J, Riquier D (1986) Complete cDNA-derived amino acid sequence of rat brown fat uncoupling protein. J Biol Chem 261:1487–1490

    Google Scholar 

  • Chen XJ, Guan MX, Clark-Walker GD (1993) MGM 101, a nuclear gene involved in maintenance of the mitochondrial genome in Saccharomyces cerevisiae. Nucleic Acids Res 21:3473–3477

    Google Scholar 

  • Cigan AM, Donahue TF (1987) Sequence and structural features associated with translational initialtor regions in yeast — a review. Gene 59:1–18

    Google Scholar 

  • Costanzo MC, Fox T (1990) Control of mitochondrial gene expression in Saccharomyces cerevisiae. Annu Rev Genet 24:91–113

    Google Scholar 

  • Daum G, Bohni PC, Schatz G (1982) Import of proteins into mitochondria. Cytochrome b2 and cytochrome c peroxidase are located in the intermembrane space of yeast mitochondria. J Biol Chem 257:13028–13033

    Google Scholar 

  • Demolis N, Mallet L, Bussereau F, Jacquet M (1993) RIM2, MSL1 and PGL1 are located within a 8-kb segment of Saccharomyces cerevisiae chromosome II which also contains the putative ribosomal gene L21 and a new putative essential gene with a leucine zipper motif. Yeast 9:645–659

    Google Scholar 

  • Foury F, Lahaye A (1987) Cloning and sequencing of the PIF1 gene involved in repair and recombination of yeast mitochondrial DNA. EMBO J 6:1441–1449

    Google Scholar 

  • Genetics Computer Group (1991) Program Manual for the GCG-Program, Version 7. Madison, Wisconsin

  • Gerbaud C, Elmerich C, Tandeau de Marsac N, Chocat P, Charpin N, Guerineau M, Aubert JP (1981) Construction of new yeast vectors and cloning of the Nif (nitrogen fixation) gene clusters of Klebsiella pneumoniae in yeast. Curr Genet 3:173–180

    Google Scholar 

  • Grivell LA (1989) Nucleo-mitochondrial interactions in yeast mitochondria. Eur J Biochem 182:477–493

    Google Scholar 

  • Hill JE, Myers AM, Koerner TJ, Tzagoloff A (1986) Yeast/E. coli shuttle vectors with multiple unique restriction sites. Yeast 2:163–167

    Google Scholar 

  • Jank B, Habermann B, Schweyen RJ, Link TA (1993) PMP47, a peroxisomal homologue of mitochondrial solute carrier proteins. Trends Biochem Sci 18:427–428

    Google Scholar 

  • Kaplan RS, Mayor JA, Wood DO (1993) The mitochondrial tricarboxylate transport proteins. cDNA cloning, primary structure, and comparison with other mitochondrial transport proteins. J Biol Chem 268:13682–13690

    Google Scholar 

  • Klingenberg M (1989) Molecular aspects of the adenine nucleotide carrier from mitochondria. Arch Biochem Biophys 270:1–14

    Google Scholar 

  • Klingenberg M (1990) Mechanism and evolution of the uncoupling protein of brown adipose tissue. Trends Biochem Sci 15:108–112

    Google Scholar 

  • Koll H, Schmidt C, Wiesenberg G, Schmelzer C (1987) Three nuclear genes suppress a yeast mitochondrial splice defect when present in high copy number. Curr Genet 12:503–509

    Google Scholar 

  • Kramer R, Palmieri F (1989) Molecular aspects of isolated and reconstituted carrier proteins from animal mitochondria. Biochim Biophys Acta, 974:1–23

    Google Scholar 

  • Kuan J, Saier MH Jr (1993) The mitochondrial carrier family of transport proteins: structural, functional and evolutionary relationships. Crit Rev Biochem Mol Biol, 28:209–233

    Google Scholar 

  • Lahaye A, Stahl H, Thines-Sempoux D, Foury F (1991) PIF1: a DNA helicase in yeast mitochondria. EMBO J 10:997–1007

    Google Scholar 

  • Lawson JE, Douglas MG (1988) Separate genes encode functionally equivalent ADP/ATP carrier proteins in Saccharomyces cerevisiae. J Biol Chem 263:14812–14818

    Google Scholar 

  • Maniatis T, Fritsh E, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York

    Google Scholar 

  • Mattick JS, Hall RM (1977) Replicative deoxyribonucleic acid synthesis in isolated mitochondria from Saccharomyces cerevisiae. J Bacteriol 130:973–982

    Google Scholar 

  • McCammon MT, Dowds CA, Orth K, Moomaw CR, Slaughter CA, Goodman JM (1990) Sorting of peroxisomal membrane protein PMP47 from Candida boidinii into peroxisomal membranes of Saccharomyces cerevisiae. J Biol Chem 265:20098–20105

    Google Scholar 

  • McCombie WR, Adams MD, Kelley JM, Fitzgerald MG, Utterback TR, Khan M, Dubnick M, Kelavage AR, Venter JC, Fields C (1992) Caenorhabditis elegans expressed sequence tags identify gene families and potential disease gene homologues. Nature Genet 1:124–131

    Google Scholar 

  • Nelson DR, Lawson JE, Klingenberg M, Douglas MG (1993) Site-directed mutagenesis of the yeast mitochondrial ADP/ATP translocator. Six arginines and one lysine are essential. J Mol Biol 230:1159–1170

    Google Scholar 

  • Pearson WR, Lipman DJ (1988) Improved tools for biological sequence comparison. Proc Natl Acad Sci USA 85:2444–2448

    Google Scholar 

  • Pfanner N, Neupert W (1990) The mitochondrial protein import apparatus. Annu Rev Biochem 59:331–353

    Google Scholar 

  • Phelps A, Schobert CT, Wohlrab H (1991) Cloning and characterization of the mitochondrial phosphate transport gene from the yeast Saccharomyces cerevisiae. Biochemistry,30:248–252

    Google Scholar 

  • Pon L, Schatz G (1991) Biogenesis of yeast mitochondria. In: Broach JR, Pringle JR, Jones EW (eds) The molecular and cellular biology of the yeast Saccharomyces. Genome dynamics, protein synthesis and energetics. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, pp 333–406

    Google Scholar 

  • Rine J (1991) Gene overexpression in studies of Saccharomyces cerevisiae. Methods Enzymol 194:239–251

    Google Scholar 

  • Romani A, Dowell E, Scarpa A (1991) Cyclic AMP-induced Mg2+ release from rat liver hepatocytes, permeabilized hepatocytes and isolated mitochondria. J Biol Chem 266:24376–24383

    Google Scholar 

  • Rothstein RJ (1983) One-step gene disruption in yeast. Methods Enzymol 101, 202–210

    Google Scholar 

  • Runswick MJ, Walker JE, Bisaccia F, Iacobazzi V, Palmieri F (1990) Sequence of the bovine 2-oxoglutarate/malate carrier protein: structural relationship to other mitochondrial transport proteins. Biochemistry 29:11033–11040

    Google Scholar 

  • Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467

    Google Scholar 

  • Schuler GD, Altschul SF, Lipman DJ (1991) A workbench for multiple alignment construction and analysis. Proteins 9:180–190

    Google Scholar 

  • Sullivan TD, Strelow LI, Illingworth CA, Phillips RL, Nelson OE (1991) Analysis of maize Brittle-1 alleles and a defective suppressor-mutator-induced mutable allele. Plant Cell 3:1337–1348

    Google Scholar 

  • Van Dyck E, Foury F, Stillman B, Brill SJ (1992) A single-stranded DNA binding protein required for mitochondrial DNA replication in S. cerevisiae is homologous to E. coli SSB. EMBO J 11:3421–3431

    Google Scholar 

  • Waldherr M, Ragnini A, Jank B, Teply R, Wiesenberger G, Schweyen RJ (1993) A multitude of suppressors of group II introns splicing defects in yeast. Curr Genet 24:301–306

    Google Scholar 

  • Wiesenberger G, Link TA, von Ahsen U, Waldherr M, Schweyen RJ (1991) MRS3 and MRS4, two suppressors of mtRNA splicing defect in yeast, are members of the mitochondrial carrier family. J Mol Biol 217:23–37

    Google Scholar 

  • Wiesenberger G, Waldherr M, Schweyen RJ (1992) The nuclear gene MRS2 is essential for the excision of group II introns from yeast mitochondrial transcripts in vivo. J Biol Chem 267:6963–6969

    Google Scholar 

  • Williamson DH, Fennel DJ (1979) Visualization of yeast mitochondrial DNA with the fluorescent stain DAPI. Methods Enzymol 56:728–733

    Google Scholar 

  • Yaffe MP (1991) Analysis of mitochondrial function and assembly. Methods Enzymol 194:627–643

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Unité de Biochimie Physiologique, Université Catholique de Louvain, Place Croix du Sud 2-20, 1348, Louvain-la-Neuve, Belgium

    Eric Van Dyck & Françoise Foury

  2. Institut für Mikrobiologie und Genetik, Vienna Biocenter, Dr. Bohrgasse 9, 1030, Vienna, Austria

    Bernhard Jank, Antonella Ragnini & Rudolf J. Schweyen

  3. Laboratoire de Bioénergétique, Institut de Chimie, University of Liège, 4000, Liege, Belgium

    Claire Duyckaerts & Francis Sluse

Authors
  1. Eric Van Dyck
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Bernhard Jank
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Antonella Ragnini
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Rudolf J. Schweyen
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Claire Duyckaerts
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Francis Sluse
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Françoise Foury
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Communicated by C.P. Hollenberg

Rights and permissions

Reprints and permissions

About this article

Cite this article

Van Dyck, E., Jank, B., Ragnini, A. et al. Overexpression of a novel member of the mitochondrial carrier family rescues defects in both DNA and RNA metabolism in yeast mitochondria. Molec. Gen. Genet. 246, 426–436 (1995). https://doi.org/10.1007/BF00290446

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00290446

Key words

Search

Navigation