Skip to main content
SpringerLink
Log in

A new member of the adenylate kinase family in yeast: PAK3 is highly homologous to mammalian AK3 and is targeted to mitochondria

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

Summary

Making use of the polymerase chain reaction primed by oligonucleotides corresponding to regions conserved between members of the nucleoside monophosphate kinase family, we have isolated the yeast gene PAK3. Pak3p belongs to the subgroup of long-form adenylate kinase isozymes (deduced molecular mass 25.3 kDa) and exhibits highest sequence similarity to bovine AK3 rather than to the yeast isozyme, Aky2p. The gene is shown to be non-essential because haploid disruption mutants are viable, both in the presence and absence of a functional AKY2 allele. It maps on chromosome V upstream of RAD3. Its expression level is low when cells are grown on glucose or other fermentable carbon sources and about threefold higher on glycerol, but can be significantly induced by ethanol. A PAK3/mouse dihydrofolate reductase fusion construct expressed in yeast is targeted to mitochondria. Transformation with PAK3 on a multicopy plasmid complements neither adenylate kinase deficiency in an aky2-disrupted yeast strain nor in Escherichia coli cells conditionally defective in adenylate kinase.

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

Abbreviations

Ap5A:

P1,P5-di(adenosine-5′)pentaphosphate

adenylate kinase:

ATP: AMP phosphotransferase (EC 2.7.4.3)

Pak3p (Aky2p):

protein product of the PAK3 (AKY2) gene

DHFR:

mouse dihydrofolate reductase

References

  • Bandlow W, Strobel G, Zoglowek C, Oechsner U, Magdolen V (1988) Yeast adenylate kinase is active simultaneously in mitochondria and cytoplasm and is required for non-fermentative growth. Eur J Biochem 178:451–457

    Google Scholar 

  • Barker WC, Hunt LT, Orcutt BC, Geroge DG, Veh LS, Chen HR, Blomquist MC, Johnson GC, Dayhoff MO (1983) Atlas of Protein Sequence and Structure, Protein Data Base, vol 7. National Biochemical Research Foundation, Washington, DC

    Google Scholar 

  • Bennetzen JL, Hall BD (1982) Codon selection in yeast. J Biol Chem 257:3026–3031

    Google Scholar 

  • Bradford M (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    Article  CAS  PubMed  Google Scholar 

  • Brune M, Schumann R, Wittinghofer F (1985) Cloning and sequencing of the adenylate kinase gene (adk) of Escherichia coli. Nucleic Acids Res 13:7139–7151

    Google Scholar 

  • Chen E, Seeburg PH (1985) Supercoil sequencing: A fast and simple method for sequencing plasmid DNA. DNA 4:165–170

    Google Scholar 

  • Chin CS, Su S, Russell PJ (1967) Adenylate kinase from bakers' yeast. I. Purification and intracellular location. Biochim Biophys Acta 132:361–369

    Google Scholar 

  • Eisenberg D (1984) Three-dimensional structure of membrane and surface proteins. Annu Rev Biochem 53:595–623

    Google Scholar 

  • Frank R, Trosin M, Tomasselli AG, Noda L, Krauth-Siegel RL, Schirmer RH (1986) Mitochondrial adenylate kinase (AK2) from bovine heart. Eur J Biochem 154:205–211

    Google Scholar 

  • Haarer BK, Lillie SH, Adams AEM, Magdolen V, Bandlow W, Brown SS (1990) Purification of profilin from Saccharomyces cerevisiae and analysis of profilin-deficient cells. J Cell Biol 110:105–114

    Google Scholar 

  • Hamilton R, Watanabe CK, de Boer HA (1987) Compilation and comparison of the sequence context around the AUG startcodon in Saccharomyces cerevisiae mRNAs. Nucleic Acids Res 15:3589–3593

    Google Scholar 

  • von Heijne G (1986) Mitochondrial targeting sequences may form amphiphilic helices. EMBO J 5:1335–1342

    Google Scholar 

  • Heil A, Muller G, Noda L, Pinder D, Schirmer H, Schirmer I, von Zabern I (1974) The amino-acid sequence of porcine adenylate kinase from skeletal muscle. Eur J Biochem 43:131–144

    Google Scholar 

  • Higgins DG, Sharp PM (1988) CLUSTAL: A package for performing multiple sequence alignment on a microcomputer. Gene 73:237–244

    Article  CAS  PubMed  Google Scholar 

  • Ito Y, Tomasselli AG, Noda LH (1980) ATP:AMP phosphotranferase from baker's yeast. Purification and properties. Eur J Biochem 105:85–92

    Google Scholar 

  • Kishi F, Tanizawa Y, Nakazawa A (1987) Isolation and characterization of two types of cDNA for mitochondrial adenylate kinase and their expression in Escherichia coli. J Biol Chem 263:11787–11789

    Google Scholar 

  • Klebe RJ, Harriss JV, Sharp ZD, Douglas MG (1983) A general method for polyethylene glycol-induced genetic transformation of bacteria and yeast. Gene 25:333–341

    Google Scholar 

  • Kuby SA, Palmieri RH, Frischat A, Fischer AH, Wu LH, Maland L, Manship M (1984) Studies on adenosine triphosphate transphosphorylases. Amino acid sequence of rabbit muscle. Biochemistry 23:2393–2399

    Google Scholar 

  • Lemire BD, Fankhauser C, Baker A, Schatz G (1989) The mitochondrial targeting function of randomly generated peptide sequences correlates with predicted helical amphiphilicity. J Biol Chem 264:20006–20015

    Google Scholar 

  • van Loon APGM, Van Eijk E, Grivell LA (1983) Biosynthesis of the ubiquinol-cytochrome c reductase complex in yeast: Discoordinate synthesis of the 11-kd subunit in response to increased gene copy number. EMBO J 2:1765–1770

    Google Scholar 

  • Magdolen V, Oechsner U, Bandlow W (1987) The complete nucleotide sequence of the gene coding for yeast adenylate kinase. Curr Genet 12:405–411

    Google Scholar 

  • Nakamura K, Nakamura A, Takamatsu H, Yoshikawa H, Yamane K (1990) Cloning and characterization of a Bacillus subtilis gene homologous to E. coli sect J Biochem (Tokyo) 107:603–607

    Google Scholar 

  • Noda LH (1973) Adenylate kinase. In: Boyer PD (ed) The Enzymes, 3rd edn, vol 8. Academic Press, New York, pp 279–305

    Google Scholar 

  • Oechsner U, Magdolen V, Zoglowek C, Hacker U, Bandlow W (1988) Yeast adenylate kinase is transcribed constitutively from a promoter in the short intergenic region to the histone H2A-1 gene. FEBS Lett 242:187–193

    Google Scholar 

  • Reynolds P, Higgins DR, Prakash L, Prakash S (1985) The nucleotide sequence of the RAD3 gene of Saccharomyces cerevisiae: a potential adenine nucleotide binding amino acid sequence and a nonessential acidic carboxyl terminal region. Nucleic Acids Res 13:2357–2372

    Google Scholar 

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

    Google Scholar 

  • Rothstein RJ (1983) One-step gene disruption in yeast. Methods Enzymol 101:202–211

    Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: A laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York

    Google Scholar 

  • Schricker R, Magdolen V, Germaier H, Bandlow W (1990) The adenylate kinase family: structure, function and open questions. In: Lachowicz TM (ed) Genetics of respiratory enzymes in yeast. Wroclaw University Press, Wroclaw, Poland, pp 172–184

    Google Scholar 

  • Schulz GE (1987) Structural and functional relationships in the adenylate kinase family. Cold Spring Harbor Symp Quant Biol 52:429–439

    Google Scholar 

  • Sherman F, Fink GR, Hicks JB (1986) Methods in yeast genetics: A laboratory manual, Revised edn. Cold Spring Harbor Laboratory Press. Cold Spring Harbor, New York

    Google Scholar 

  • Spurgin P, Tomasselli AG, Schiltz E (1989) The amino acid sequence of adenylate kinase from Paracoccus denitrificans and its relationship to mitochondrial and microbial adenylate kinases. Eur J Biochem 179:621–628

    CAS  PubMed  Google Scholar 

  • Strobel G, Magdolen V, Oechsner U, Huh H-S, Bandlow W (1988) The 5′-upstream region of the yeast 25S rRNA gene contains a promoter element allowing expression in yeast and E. coli. Curr Genet 14:293–302

    Google Scholar 

  • Struhl K (1985) Nucleotide sequence and transcriptional mapping of the yeast pet56-his3-ded1 gene region. Nucleic Acids Res 13:8587–8601

    Google Scholar 

  • Stueber D, Ibrahimi I, Cutler D, Dobberstein B, Bujard H (1984) A novel in vitro transcription-translation system: accurate and efficient synthesis of single proteins from cloned DNA sequences. EMBO J 3:3143–3148

    Google Scholar 

  • Tomasselli AG, Schirmer RH, Noda LH (1979) Mitochondrial GTP-AMP Phosphotransferase. 1. Purification and properties. Eur J Biochem 93:257–262

    Google Scholar 

  • Tomasselli AG, Mast E, Janes W, Schiltz E (1986a) The complete amino acid sequence of adenylate kinase from baker's yeast. Eur J Biochem 155:111–119

    Google Scholar 

  • Tomasselli AG, Frank R, Schulz GE (1986b) The complete primary structure of GTP:AMP phosphotransferase from beef heart mitochondria. FEBS Lett 202:303–308

    Google Scholar 

  • Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications. Proc Natl Acad Sci USA 76:4350–4354

    CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Laboratorium für Molekulare Zellbiologie und Allgemeine Pathologie, Institut für Pathologie und Rechtsmedizin der Universität Ulm, Neuherbergstrasse 11, W-8000, München 45, Germany

    Roland Schricker

  2. Institut für Genetik und Mikrobiologie der Universität München, Maria-Ward-Strasse la, W-8000, München 19, Germany

    Viktor Magdolen & Wolfhard Bandlow

Authors
  1. Roland Schricker
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Viktor Magdolen
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Wolfhard Bandlow
    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

Schricker, R., Magdolen, V. & Bandlow, W. A new member of the adenylate kinase family in yeast: PAK3 is highly homologous to mammalian AK3 and is targeted to mitochondria. Molec. Gen. Genet. 233, 363–371 (1992). https://doi.org/10.1007/BF00265432

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00265432

Key words

Search

Navigation