Skip to main content

Transcription of the yeast mitochondrial genome requires cyclic AMP

Molecular and General Genetics MGG volume 241, pages 213–224 (1993)Cite this article

Abstract

Using various mutant strains and nutritional manipulations, we investigated a potential role for cyclic AMP (cAMP) in the regulation of mitochondrial (mt) gene expression in the yeast Saccharomyces cerevisiae. In RAS mutants known to have either abnormally low or high cellular levels of this nucleotide, we show that both mt transcription rate and overall mt transcript levels vary directly with cellular cAMP levels. We further show that nutritional downshift of actively growing cells causes a severe, rapid fall in cAMP levels, and that this fall is concomitant with the stringent mt transcriptional curtailment that we and others have previously shown to follow this nutritional manipulation. In in vitro mt transcription assays using intact organelles from downshifted and actively growing cells, stringently curtailed mt gene expression can be restored to 75% of control levels by addition of cAMP to the assay mix. Consistent with these observations a RAS2 vall9mutant strain, which cannot adjust cAMP levels in response to external stimuli, shows no mt stringent response following nutritional downshift. We also demonstrate a significant but transient increase in both mt transcript levels and mt transcription rate following shift of actively respiring wild-type cells to glucose-based medium, a manipulation known to cause a short-lived pulse of cAMP in yeast; similar manipulation of the RAS2 vall9mutant strain generates no such response. Taken together all these observations indicate that cellular cAMP levels are involved in the regulation of mt transcription in yeast. Moreover, the lack of a mt stringent transcriptional response following downshift in a strain in which the BCY1 gene had been insertionally inactivated suggests that cAMP may influence mt transcription via a mt cAMP-dependent protein kinase. These results link mt gene expression with mechanisms governing growth control and nutrient adaptation in yeast, and they provide a means by which nit gene expression might be coordinated with that of related nuclear genes.

This is a preview of subscription content, access via your institution.

Access options

Buy single article

Instant access to the full article PDF.

43,34 €

Price includes VAT (Finland)
Tax calculation will be finalised during checkout.

References

  • Bandlow W, Schwarz U, Rodel G, Strobel G, Wachter C (1985) Isolation of a CAMP receptor protein from yeast mitochondria (Mr 45000) and comparison with mitochondrial RNA polymerase (Mr 45000). Biol Chem Hoppe-Seyler 366:545–553

    Google Scholar 

  • Birky CW (1975) Effects of glucose repression on the transmission of mitochodrial genes in yeast (Saccharomyces cerevisiae). Genetics 80:695–709

    Google Scholar 

  • Black RJ, Friedman RM (1989) Cytokines and oncogene activity. Cancer Surv 8:725–739

    Google Scholar 

  • Breviario D, Hinnebusch A, Cannon J, Tatchell K, Dhar R (1986) Carbon source regulation of RAS1 expression in Saccharomyces cerevisiae and the phenotype of ras2 − cells. Proc Natl Acad Sci USA 83:4152–4156

    Google Scholar 

  • Breviario D, Hinnebusch AG, Dhar R (1988) Multiple regulatory mechanisms control the expression of the RAS1 and RAS2 genes of Saccharomyces cerevisiae. EMBO J 7:1805–1813

    Google Scholar 

  • Broach JR (1991) Ras-regulated signaling processes in Saccharomyces cerevisiae. Curr Opin Genet Dev 1:370–377

    Google Scholar 

  • Broach JR, Deschenes RJ (1990) The function of ras genes in Saccharomyces cerevisiae. Adv Cancer Res 54:79–139

    Google Scholar 

  • Broek D (1989) Eukaryotic RAS proteins and yeast proteins with which they interact. Curr Top Microbiol Immunol 147:155–169

    Google Scholar 

  • Broek D, Toda T, Michaeli T, Levin L, Birchmeier C, Zoller M, Powers S, Wigler M (1987) The S. cerevisiae CDC25 gene product regulates the RAS/adenylate cyclase pathway. Cell 48:789–799

    Google Scholar 

  • Camonis J, Kalekine M, Gondre B, Garreau H, Boy-Marcotte E, Jacquet M (1986) Characterization, cloning, and sequence analysis of the CDC25 gene which control the cyclic AMP level of Saccharomyces cerevisiae. EMBO J 5:375–380

    Google Scholar 

  • Cannon JF, Gitan R, Tatchell K (1990) Yeast cAMP-dependent protein kinase regulatory subunit mutations display a variety of phenotypes. J Biol Chem 265:11897–11904

    Google Scholar 

  • Cantwell R, McEntee CM, Hudson AP (1992) Regulation of mitochondrial transcription during the stringent response in yeast. Curr Genet 21:241–247

    Google Scholar 

  • Chandrasekaran K, Jayaraman J (1978) Effect of cyclic AMP on the biogenesis of cytochrome oxidase in yeast. FEBS Lett 87:52–54

    Google Scholar 

  • Daniel J, Becker JM, Enari E, Levitzki A (1987) The activation of adenylate cyclase by guanyl nucleotides in Saccharomyces cerevisiae is controlled by the CDC25 start gene product. Mol Cell Biol 7:3857–3861

    Google Scholar 

  • de Crombrugghe B, Busby S, Buc H (1984) Cyclic AMP receptor protein: role in transcriptional activation. Science 224:831–838

    Google Scholar 

  • Defeo-Jones D, Scolnick E, Koller R, Dhar R (1983) ras-related gene sequence identified from Saccharomyces cerevisiae. Nature 306:707–709

    Google Scholar 

  • Fang M, Butow RA (1970) Nucleotide reversal of mitochondrial repression in Saccharomyces cerevisiae. Biochem Biophys Res Commun 41:1579–1583

    Google Scholar 

  • Gibbs JB, Marshall MS (1989) The ras oncogene-an important regulatory element in lower eukaryotic organisms. Microbiol Rev 53:171–185

    Google Scholar 

  • Glaichenhaus N, Leopold P, Cuzin F (1986) Increased levels of mitochondrial gene expression in rat fibroblast cell immortalized or transformed by viral and cellular oncogenes. EMBO J 5:1261–1265

    Google Scholar 

  • Groot GFP, van Harten-Loosbroeck N, van Ommen G-JB, Pijst HLA (1980) RNA synthesis in isolated yeast mitochondria. Nucleic Acids Res 9:6369–6377

    Google Scholar 

  • Hixson CS, Krebs EG (1980) Characterization of a cyclic AMP binding protein from bakers' yeast. J Biol Chem 255:2137–2145

    Google Scholar 

  • Hudspeth MES, Shumard DS, Tatti KM, Grossman L (1980) Rapid purification of yeast mitochondrial DNA in high yield. Biochim Biophys Acta 610:221–228

    Google Scholar 

  • Ito H, Fukuda Y, Murata K, Kimura A (1983) Transformation of intact yeast cells treated with alkali cations. J Bacteriol 153:163–168

    Google Scholar 

  • Kief DR, Warner JR (1981) Coordinate control of ribosomal nucleic acids and ribosomal proteins during nutritional shift-up in Saccharomyces cerevisiae. Mol Cell Biol 1:1007–1015

    Google Scholar 

  • Mahler HR, Lin CC (1978) Exogenous adenosine 3′,5′-monophosphate can release yeast from catabolite repression. Biochem Biophys Res Commun 83:1039–1047

    Google Scholar 

  • Marshall MS, Gibbs JB, Scolnick EM, Sigal IS (1987) Regulatory function of the Saccharomyces cerevisiae RAS C-terminus. Mol Cell Biol 7:2309–2315

    Google Scholar 

  • Marshall MS, Gibbs JB, Scolnick EM, Sigal IS (1988) An adenylate cyclase from Saccharomyces cerevisiae that is stimulated by RAS proteins with effector mutations. Mol Cell Biol 8:52–61

    Google Scholar 

  • Marshall MS, Hill WS, Ng AS, Vogel US, Schaber MD, Scolnick EM, Dixon RAF, Sigal IS, Gibbs JB (1989) A C-terminal domain of GAP is sufficient to stimulate ras p21 GTPase activity. EMBO J 8:1105–1110

    Google Scholar 

  • Matsumoto K, Uno I, Oshima Y, Ishikawa Y (1982) Isolation and characterization of yeast mutants deficient in adenylate cyclase and cAMP-dependent protein kinase. Proc Natl Acad Sci USA 79:2355–2359

    Google Scholar 

  • Matsumoto K, Uno I, Hikawa T, Oshima Y (1983) Cyclic AMP may not be involved in catabolite repression is Saccharomyces cerevisiae: evidence from mutants unable to synthesize it. J Bacteriol 156:898–900

    Google Scholar 

  • Mbonyi K, Beullens M, Detremerie K, Geerts L, Thevelein JM (1988) Requirement of one functional RAS gene and inability of an oncogenic ras variant to mediate the glucose-induced cyclic AMP signal in the yeast Saccharomyces cerevisiae. Mol Cel Biol 8:3051–3057

    Google Scholar 

  • Mbonyi K, van Aelst L, Arguelles JC, Jans AWH, Thevelein JM (1990) Glucose-induced hyperaccumulation of cyclic AMP and defective glucose repression in yeast strains with reduced activity of cyclic AMP-dependent protein kinase. Mol Cell Biol 10:4518–4523

    Google Scholar 

  • McEntee CM, Hudson AP (1989) Preparation of RNA from unspheroplasted yeast cells (Saccharomyces cerevisiae). Anal Biochem 176:303–306

    Google Scholar 

  • McEntee CM, Cantwell R, Thomas LC, Hudson AP (1989) Mitochondrial rRNA-containing petite strains of yeast (Saccharomyces cerevisiae) show a normal nuclear-mitochondrial stringent response. Biochem Biophys Res Commun 164:362–369

    Google Scholar 

  • Merino A, Buckbinder L, Mermelstein FH, Reinberg D (1989) Phosphorylation of cellular proteins regulates their binding to the cAMP response element. J Biol Chem 264:21266–21276

    Google Scholar 

  • Mueller DM, Getz G (1986) Steady state analysis of mitochondrial RNA after growth of yeast S. cerevisiae under catabolite repression and derepression. J Biol Chem 261:11816–11822

    Google Scholar 

  • Muller G, Bandlow W (1987) Cyclic AMP-dependent protein kinase activity in yeast mitochondria. Z Naturforsch 42c:1291–1302

    Google Scholar 

  • Muller G, Bandlow W (1989a) An amphitropic cAMP-binding protein in yeast mitochondria. 1. Synergistic control of the intramitochondrial location by calcium and phospholipid. Biochemistry 28:9957–9967

    Google Scholar 

  • Muller G, Bandlow W (1989b) An amphitropic cAMP-binding protein in yeast mitochondria. 2. Phospholipid nature of the membrane anchor. Biochemistry 28:9968–9973

    Google Scholar 

  • Muller G, Bandlow W (1989c) An amphitropic cAMP-binding protein in yeast mitochondria. 3. Membrane release requires both Ca2+-dependent phosphorylation of the CAMP-binding protein and a phospholipid-activated mitochondrial phospholipase. Biochemistry 28:9974–9981

    Google Scholar 

  • Powers S, Kataoka T, Fasano O, Goldfarb M, Strathern JN, Broach JR, Wigler M (1984) Genes in S. cerevisiae encoding proteins with domains homologous to the mammalian ras proteins. Cell 36:607–612

    Google Scholar 

  • Ray DB, Butow RA (1979a) Regulation of mitochondrial ribosomal RNA synthesis in yeast. I. In search of a relaxation of stringency. Mol Gen Genet 173:227–238

    Google Scholar 

  • Ray DB, Butow RA (1979b) Regulation of mitochondrial ribosomal RNA synthesis in yeast. II. Effects of temperature sensitive mutants defective in cytoplasmic protein synthesis. Mol Gen Genet 173:239–247

    Google Scholar 

  • Richtsmeier WJ, Grossberg SE (1989) Inhibitory effects of mitochondrial metabolic inhibitors on interferon action. J Interferon Res 9:87–96

    Google Scholar 

  • Rigby PW, Dieckman M, Rhodes C, Berg P (1977) Labeling of deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol 113:237–251

    Google Scholar 

  • Robinson LC, Gibbs JB, Marshall MS, Sigal IS, Tatchell K (1987) CDC25: a component of the RAS-adenylate cyclase pathway in Saccharomyces cerevisiae. Science 235:1218–1221

    Google Scholar 

  • Robinson-Steiner AM, Beebe SJ, Rannels SR, Corbin JD (1984) Microheterogeneity of type II cAMP-dependent protein kinase in various mammalian species and tissues. J Biol Chem 259:10596–10605

    Google Scholar 

  • Rodel G, Muller G, Bandlow W (1985) Cyclic AMP receptor protein from yeast mitochondria: submitochondrial localization and preliminary characterization. J Bacteriol 161:7–12

    Google Scholar 

  • Roesler WJ, Vandenbark GR, Hanson RW (1988) Cyclic AMP and the induction of eukaryotic transcription. J Biol Chem 263:9063–9066

    Google Scholar 

  • Shan B, Vazquez E, Lewis JA (1990) Interferon selectively inhibits the expression of mitochondrial genes: a novel pathway for interferon-mediated responses. EMBO J 9:4307–4314

    Google Scholar 

  • Sherman F, Fink GR, Hicks JB (1983) Methods in yeast genetics, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York

    Google Scholar 

  • Tanaka T, Matsumoto K, Toh-e A (1989) IRA1, an inhibitory regulator of the RAS-cyclic AMP pathway in Saccharomyces cerevisiae. Mol Cell Biol 9:757–768

    Google Scholar 

  • Tanaka K, Nakafuku M, Tamanoi F, Kaziro Y, Matsumoto K, Toh-e A (1990) IRA2, a second gene of Saccharomyces cerevisiae that encodes a protein with a domain homologous to mammalian ras GTPase-activating protein. Mol Cell Biol 10:4303–4313

    Google Scholar 

  • Tatchell K (1986) RAS genes and growth control in Saccharomyees cerevisiae. J Bacteriol 166:364–367

    Google Scholar 

  • Tatchell K, Robinson LC, Breitenbach M (1985) RAS2 of Saccharomyces cerevisiae is required for gluconeogenic growth and proper response to nutrient limitation. Proc Natl Acad Sci USA 82:3785–3789

    Google Scholar 

  • Taylor SS, Buechler JA, Yonemoto W (1990) cAMP-dependent protein kinase: framework for a diverse family of regulatory enzymes. Annu Rev Biochem 59:971–1005

    Google Scholar 

  • Thomas PS (1980) Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci USA 77:5201–5205

    Google Scholar 

  • Toda T, Uno I, Ishikawa T, Powers S, Kataoka T, Broek D, Cameron S, Broach JR, Matsumoto K, Wigler M (1985) In yeast, RAS proteins are controlling elements of adenylate cyclase. Cell 40:27–36

    Google Scholar 

  • Toda T, Cameron S, Sass P, Zoller M, Scott JD, McMullen B, Hurwitz M, Krebs EG, Wigler M (1987a) Cloning and characterization of BCY1, a locus encoding a regulatory subunit of the cyclic AMP-dependent protein kinase in Saccharomyces cerevisiae. Mol Cell Biol 7:1371–1377

    Google Scholar 

  • Toda T, Cameron S, Sass P, Zoller M, Wigler M (1987b) Three different genes in S. cerevisiae encode the catalytic subunits of the cAMP-dependent protein kinase. Cell 50:277–287

    Google Scholar 

  • Van Wijk R, Konijn TM (1971) Cyclic 3′,5′ AMP in Saccharomyces cerevisiae under various conditions of catabolite repression. FEBS Lett 13:184–186

    Google Scholar 

  • Warner JR (1982) The yeast ribosome: structure, function, and synthesis. In: Strathern JN, Jones JN, Broach JR (eds) The molecular biology of the yeast Saccharomyces. Metabolism and gene expression. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, pp 529–560

    Google Scholar 

  • Warner JR, Gorenstein C (1978) Yeast has a true stringent response. Nature 275:338–339

    Google Scholar 

  • Watanabe H, Chisaka T, Higuchi T, Tanaka A, Horii Y, Sugimoto T, Imanishi J (1989) Effect of human interferons on morphological differentiation and suppression of N-myc expression in human neuroblastoma cells. Jpn J Cancer Res 80:1072–1076

    Google Scholar 

  • Yamamoto KK, Gonzalez GA, Biggs WH, Montminy MR (1988) Phosphorylation-induced binding and transcriptional efficacy of nuclear factor CREB. Nature 334:494–498

    Google Scholar 

  • Zassenhaus HP, Butow RA, Hannon YP (1982) Rapid electroelution of nucleic acids from agarose and acrylamide gels. Anal Biochem 125:125–130

    Google Scholar 

  • Zitomer RS, Nichols DS (1978) Kinetics of repression of yeast cytochrome c. J Bacteriol 135:39–44

    Google Scholar 

Download references

Author information

Author notes

  1. Catherine M. McEntee

    Present address: Dept. Molecular Biology, Lewis Thomas Laboratory, Princeton University, 08544, Princeton, NJ, USA

Affiliations

  1. Dept. Microbiology and Immunology, The Medical College of Pennsylvania, 3300 Henry Avenue, 19129, Philadelphia, PA, USA

    Catherine M. McEntee, Mahboob U. Rahman & Alan P. Hudson

  2. Research Service, Dept. Veterans Affairs Medical Center, University and Woodland Avenues, 19104, Philadelphia, PA, USA

    Robin Cantwell & Alan P. Hudson

Authors
  1. Catherine M. McEntee
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Robin Cantwell
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Mahboob U. Rahman
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Alan P. Hudson
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Communicated by D. Lonsdale

Rights and permissions

Reprints and Permissions

About this article

Cite this article

McEntee, C.M., Cantwell, R., Rahman, M.U. et al. Transcription of the yeast mitochondrial genome requires cyclic AMP. Molec. Gen. Genet. 241, 213–224 (1993). https://doi.org/10.1007/BF00280219

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00280219

Key words

  • Saccharomyces cerevisiae
  • Mitochondria
  • Transcriptional regulation
  • Protein phosphorylation
  • Stringent response