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Molecular cloning of the yeast OPI3 gene as a high copy number suppressor of the cho2 mutation

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Abstract

By functional complementation of the auxotrophic requirements for choline of a cdg1, cho2 double-mutant, by transformation with a genomic DNA library in a high copy number plasmid, two different types of complementing DNA inserts were identified. One type of insert was earlier shown to represent the CHO2 structural gene. In this report we describe the molecular and biochemical characterization of the second type of complementing activity. The transcript encoded by the cloned gene was about 1000-nt in length and was regulated in response to the soluble phospholipid precursors, inositol and choline. A gene disruption resulted in no obvious growth phenotype at 23°C or 30°C, but in a lack of growth at 37°C in the presence of monomethylethanolamine. Null-mutants exhibited an inositol-secretion phenotype, indicative of mutations in the lipid biosynthetic pathway. Complementation analysis, biochemical analysis of the phospholipid methylation pathway in vivo, and comparison of the restriction pattern of the cloned gene to published sequences, unequivocally identified the cloned gene as the OPI3 gene, encoding phospholipid-N-methyltransferase in yeast. When present in multiple copies the OPI3 gene efficiently suppresses the phospholipid methylation defect of a cho2 mutation. As a result of impaired synthesis of phosphatidylcholine, the INO1-deregulation phenotype is abolished in cho2 mutants transformed with the OPI3 gene on a high copy number plasmid. Taken together, these data demonstrate a significantly overlapping specificity of the OPI3 gene product for three sequential phospholipid methylation reactions in the de novo Ptd-Cho biosynthetic pathway.

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References

  • Atkinson K, Vogel S, Henry SA (1980) J Biol Chem 255:6653–6661

    Google Scholar 

  • Bailis AM, Poole MA, Carman GM, Henry SA (1987) Mol Cell Biol 7:167–176

    Google Scholar 

  • Carman GM, Henry SA (1989) Annu Rev Biochem 58:635–669

    Google Scholar 

  • Elion EA, Warner JR (1984) Cell 39:663–673

    Google Scholar 

  • Folch J, Lees M, Sloan-Stanley GH (1957) J Biol Chem 264:2945–2950

    Google Scholar 

  • Gaynor PM, Gill T, Toutenhoofd S, Summers EF, McGraw P, Homann MJ, Henry SA, Carman GM (1991) Biochim Biophys Acta 1090:326–332

    Google Scholar 

  • Greenberg ML, Reiner B, Henry SA (1982) Genetics 100:19–33

    Google Scholar 

  • Hill JE, Myers AM, Koerner TJ, Tzagoloff A (1986) Yeast 2:163–167

    Google Scholar 

  • Hirsch JP, Henry SA (1986) Mol Cell Biol 6:3320–3328

    Google Scholar 

  • Holm C, Meeks-Wagner DW, Fangman WL, Botstein D (1986) Gene 42:169–173

    Google Scholar 

  • Klig LS, Homann MJ, Kohlwein SD, Kelley MJ, Henry SA, Carman GM (1988) J Bacteriol 170:1878–1886

    Google Scholar 

  • Kodaki T, Yamashita S (1987) J Biol Chem 262:15428–15435

    Google Scholar 

  • Kodaki T, Hosaka K, Nikawa J, Yamashita S (1991) J Biochem 109:276–287

    Google Scholar 

  • Kuchler K, Daum G, Paltauf F (1986) J Bacteriol 165:901–911

    Google Scholar 

  • McGraw P, Henry SA (1989) Genetics 122:317–330

    Google Scholar 

  • Sherman F, Fink GR, Hicks JB (1979) Cold Spring Harbor Laboratory, Cold Spring Harbor, New York

  • Summers EF, Letts VA, McGraw P, Henry SA (1988) Genetics 120:909–922

    Google Scholar 

  • Zinser E, Sperka-Gottlieb CDM, Fasch E.-V., Kohlwein SD, Paltauf F, Daum G (1991) J Bacteriol 173:2026–2034

    Google Scholar 

Download references

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Communicated by R. J. Schweyen

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Preitschopf, W., Lückl, H., Summers, E. et al. Molecular cloning of the yeast OPI3 gene as a high copy number suppressor of the cho2 mutation. Curr Genet 23, 95–101 (1993). https://doi.org/10.1007/BF00352006

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  • DOI: https://doi.org/10.1007/BF00352006

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