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Cloning and functional characterization of a fatty acid synthase component FAS2 gene from Saccharomyces kluyveri

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Abstract

A gene coding the α subunit of fatty acid synthase (FAS2) was isolated from the budding yeast Saccharomyces kluyveri. Nucleotide sequence analysis indicated that this gene, termed Sk-FAS2, coded a protein having an amino acid sequence 83% identical to the FAS2 protein of S. cerevisiae (Sc-FAS2). The Sk-FAS2 gene was able to functionally complement an S. cerevisiae fas2 disruptant. This Sk-FAS2-expressing strain was found to produce larger amounts of C18 than C16, in contrast to the Sc-FAS2-expressing fas2 strain. In addition, fusion genes of Sk-FAS2 and Sc-FAS2 were transformed into a fas2-disrupted strain of S. cerevisiae, and fatty acid analysis of these transformants suggested that the region containing the acyl carrier protein and β-ketoacyl reductase domains of yeast FAS2 protein play an important role in determining carbon chain length of fatty acids.

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References

  • Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K (eds) (1992) Current protocols in molecular biology. Green Publishing Associates and Wiley-Interscience, Wiley, New York

  • Bloch K, Vance D (1977) Control mechanisms in the synthesis of saturated fatty acids. Annu Rev Biochem 46:263–298

    Article  CAS  PubMed  Google Scholar 

  • Burke D, Dawson D, Stearns T (eds) (2000) Methods in yeast genetics: a cold spring harbor laboratory manual, 2000 edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor

    Google Scholar 

  • Chirala SS, Kuziora MA, Spector DM, Wakil SJ (1987) Complementation of mutants and nucleotide sequence of FAS1 gene encoding β subunit of yeast fatty acid synthase. J Biol Chem 262:4231–4240

    CAS  PubMed  Google Scholar 

  • Fichtlscherer F, Wellein C, Mittag M, Schweizer E (2000) A novel function of yeast fatty acid synthase. Subunit α is capable of self-pantetheinylation. Eur J Biochem 267:2666–2671

    Article  CAS  PubMed  Google Scholar 

  • Ito Y, Oh-hashi Y, Okuyama H (1986) Fatty acid synthetase as a thermoreceptor in Candida utilis. J Biochem 99:1713–1718

    CAS  PubMed  Google Scholar 

  • Jones JS, Prakash L (1990) Yeast Saccharomyces cerevisiae selectable markers in pUC18 polylinkers. Yeast 6:363–369

    Article  CAS  PubMed  Google Scholar 

  • Kajiwara S, Aritomi T, Suga K, Ohtaguchi K, Kobayashi O (2000) Overexpression of the OLE1 gene enhances ethanol fermentation by Saccharomyces cerevisiae. Appl Microbiol Biotechnol 53:568–574

    Article  CAS  PubMed  Google Scholar 

  • Kajiwara S, Oura T, Shishido K (2001) Cloning of a fatty acid synthase component FAS1 gene from Saccharomyces kluyveri and its functional complementation of S. cerevisiae fas1 mutant. Yeast 18:1339–1345

    Article  CAS  PubMed  Google Scholar 

  • Kaneko Y, Rueksomtawin K, Maekawa T, Harashima S (2003) Structural and functional analysis of the FAS1 gene encoding fatty acid synthase beta-subunit in methylotrophic yeast Hansenula polymorpha. Advanced research on plant lipids. Kluwer, Dordrecht, pp 61–64

    Google Scholar 

  • Kötig H, Rottner G, Beck K-F, Schweizer M, Schweizer E (1991) The pentafunctional FAS1 genes of Saccharomyces cerevisiae and Yarrowia lipolytica are co-linear and considerably longer than previously estimated. Mol Gen Genet 226:310–314

    Article  Google Scholar 

  • Ming D, Kong Y, Lambert MA, Huang Z, Ma J (2002a) How to describe protein motion without amino acid sequence and atomic coordinates. Proc Natl Acad Sci USA 99:8620–8625

    Article  CAS  Google Scholar 

  • Ming D, Kong Y, Wakil SJ, Brink J, Ma J (2002b) Domain movements in human fatty acid synthase by quantized elastic deformational model. Proc Natl Acad Sci USA 99:7895–7899

    Article  CAS  Google Scholar 

  • Mohamed AH, Chirala SS, Mody NH, Huang WY, Wakil SJ (1988) Primary structure of the multifunctional alpha subunit protein of yeast fatty acid synthase derived from FAS2 gene sequence. J Biol Chem 263:12315–12325

    CAS  PubMed  Google Scholar 

  • Oh C-S, Toke DA, Mandala S, Martin CE (1997) ELO2 and ELO3, homologues of the Saccharomyces cerevisiae ELO1 gene, function in fatty acid elongation and are required for sphingolipid formation. J Biol Chem 272:17376–17384

    Article  CAS  PubMed  Google Scholar 

  • Okuyama H, Saito M (1979) Regulation by temperature of the chain length of fatty acids in yeast. J Biol Chem 254:12281–12284

    CAS  PubMed  Google Scholar 

  • Ratledge C, Evans CT (1991) Lipids and their metabolism. In: Rose AH, Harrison JS (eds) The Yeast, 2nd edn, vol 4. Academic, London, pp 367–455

  • Rössler H, Rieck C, Delong T, Hoja U, Schweizer E (2003) Functional differentiation and selective inactivation of multiple Saccharomyces cerevisiae genes involved in very-long-chain fatty acid synthesis. Mol Genet Genomics 269:290–298

    PubMed  Google Scholar 

  • Saitoh S, Takahashi K, Nabeshima K, Yamashita Y, Nakaseko Y, Hirata A, Yanagida M (1996) Aberrant mitosis in fission yeast mutants defective in fatty acid synthetase and acetyl CoA carboxylase. J Cell Biol 134:949–961

    Article  CAS  PubMed  Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (eds) (1989) Molecular cloning: a labortory manual, 2nd edn, Cold Spring Harbor Laboratory Press, Cold Spring Harbor

  • Schjerling CK, Hummel R, Hansen JK, Børsting C, Mikkelsen JM, Kristiansen K Knudsen J (1996) Disruption of the gene encoding the acyl–CoA-binding protein (ACB1) perturbs acyl-CoA metabolism in Saccharomyces cerevisiae. J Biol Chem 271:22514–22521

    Article  CAS  PubMed  Google Scholar 

  • Schneiter R, Guerra CE, Lampl M, Tatzer V, Zellnig G, Klein HL, Kohlwein SD (2000a) A novel cold-sensitive allele of the rate-limiting enzyme of fatty acid synthesis, acetyl coenzyme A carboxylase, affects the morphology of the yeast vacuole through acylation of Vac8p. Mol Cell Biol 20:2984–2995

    Article  CAS  Google Scholar 

  • Schneiter R, Tatzer V, Gogg G, Leitner E, Kohlwein SD (2000b) Elo1p-dependent carboxy-terminal elongation of C14:1Delta(9) to C16:1Delta(11) fatty acids in Saccharomyces cerevisiae. J Bacteriol 182:3655–3660

    Article  CAS  Google Scholar 

  • Schweizer M, Roberts LM, Hötke H-J, Takabayashi K, Hölerer E, Hoffmann B, Müler G, Kötig H, Schweizer E (1986) The pentafunctional FAS1 gene of yeast: its nucleotide sequence and order of the catalytic domains. Mol Gen Genet 203:479–486

    Article  CAS  PubMed  Google Scholar 

  • Schweizer E, Müler G, Roberts LM, Schweizer M, Röch J, Wiesner P, Beck J, Stratmann D, Zauner I (1987) Genetic control of fatty acid synthetase biosynthesis and structure in lower fungi. Fat Sci Technol 89:570–577

    Google Scholar 

  • Schweizer E, Kötig H, Regler R, Rottner G (1988) Genetic control of Yarrowia lipolytica fatty acid synthetase biosynthesis and function. J Basic Microbiol 28:283–292

    Article  CAS  PubMed  Google Scholar 

  • Southard SB, Cihlar RL (1995) Analysis and expression of the Candida albicans FAS2 gene. Gene 156:133–138

    Article  CAS  PubMed  Google Scholar 

  • Suutari M, Laakso S (1994) Microbial fatty acids and thermal adaptation. Crit Rev Microbiol 20:285–328

    Article  CAS  PubMed  Google Scholar 

  • Toke DA, Martin CE (1996) Isolation and characterization of a gene affecting fatty acid elongation in Saccharomyces cerevisiae. J Biol Chem 271:18413–18422

    Article  CAS  PubMed  Google Scholar 

  • Trotter PJ (2001) The genetics of fatty acid metabolism in Saccharomyces cerevisiae. Annu Rev Nutr 21:91–119

    Article  Google Scholar 

  • Wakil SJ (1989) Fatty acid synthase, a proficient multifunctional enzyme. Biochemistry 28:4523–4530

    Article  CAS  PubMed  Google Scholar 

  • Wakil SJ, Stoops JK, Joshi VC (1983) Fatty acid synthesis and its regulation. Annu Rev Biochem 52:537–579

    Article  CAS  PubMed  Google Scholar 

  • Watanabe K, Oura T, Sakai H, Kajiwara S (2004) Yeast Δ12 fatty acid desaturase: gene cloning, expression and function. Biosci Biotechnol Biochem 68:721–727

    Article  CAS  PubMed  Google Scholar 

  • Zhao X-J, Cihlar RL (1994) Isolation and sequence of the Candida albicans FAS1 gene. Gene 147:119–124

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This work was financed by Grants for Young Scientists (no. 10760045 and no. 13760061) from the Japan Society for the Promotion of Science.

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Authors and Affiliations

  1. Department of Life Science, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 4259-B5 Nagatsuta, Midori-ku, 226-8501, Yokohama, Kanagawa, Japan

    Takahiro Oura & Susumu Kajiwara

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  1. Takahiro Oura
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  2. Susumu Kajiwara
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Correspondence to Susumu Kajiwara.

Additional information

Communicated by S. Hohmann

GenBank Submission: The nucleotide sequences reported in this paper have been submitted to GenBank Nucleotide Sequence Database under the accession number AB115969.

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Oura, T., Kajiwara, S. Cloning and functional characterization of a fatty acid synthase component FAS2 gene from Saccharomyces kluyveri . Curr Genet 49, 393–402 (2006). https://doi.org/10.1007/s00294-006-0063-4

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  • DOI: https://doi.org/10.1007/s00294-006-0063-4

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