Abstract
A mutant (plc1-1) of Schizosacharomyces pombe unable to grow on a minimal medium containing high amounts of phosphate was selected. On yeast-extract agar its growth is temperature sensitive. Tests in liquid synthetic medium show that growth of the mutant is partially restored by lowering the phosphate and inositol concentrations in the growth medium. The growth defect is fully suppressed by a plasmid encoding a putative protein having the structural features of phosphoinositide-specific phospholipases C (PI-PLC). This protein, of 899 amino-acids, contains the characteristic X and Y domains found in all PI-PLCs of higher and lower eucaryotes and reveals, in addition, an EF-hand motif (putative Ca2+-binding site). Like the corresponding enzyme from Saccharomyces cerevisiae, the S. pombe PI-PLC is most similar to the δ form of PI-PLC isoenzymes. The cloned gene integrates at the plc1 site indicating that plc1 codes for a putative PI-PLC. Plc1 physically maps on the left arm of chromosome II between rad11 and mei3.
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References
Alfa C, Fantes P, Hyams J, McLeod M, Warbrick E (1993) Experiments with fission yeast. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York
Barbet N, Muriel WJ, Carr AM (1992) Versatile shuttle vectors and genomic libraries for use with Schizosaccharomyces pombe. Gene 114:59–66
Berridge MJ (1993) Inositol triphosphate and calcium signalling. Nature 361:315–325
Bullock WO, Fernandez JM, Short JM (1987) XL1-Blue. A high-efficiency plasmid transforming recA Escherichia coli strain with beta-galactosidase selection. Bio Techniques 5:371–377
Dhamija SS, Fluri R, Schweingruber ME (1987) Two genes control three alkaline phosphatases in Schizosaccharomyces pombe. Curr Genet 11:467–473
Drayer AL, Van der Kaay J, Mayr GW, Van Haastert PJM (1994) Role of phospholipase C in Dictyostelium: formation of inositol 1,4,5-triphosphate and normal development in cells lacking phospholipase C activity. EMBO J 13:1601–1609
Flick JS, Thorner J (1993) Genetic and biochemical characterization of a phosphatidylinositol-specific phospholipase C in Saccharomyces cerevisiae. Mol Cell Biol 13:5861–5876
Genetics Computer Group (1994) Program manual for the GCG package, version 8.0, September 1994. 575 Science Drive, Madison, Wisconsin, USA
Gutz H, Leslot H, Leupold U, Loprieno N (1974) Schizosaccharomyces pombe In: King RC (ed) Handbook of genetics, Vol 1. Plenum Press, New York, pp 395–446
Hoheisel JD, Maier E, Mott R, McCarthy L, Grigoriev AV, Schalkwyk LC, Nizetic D, Francis F, Lehrach H (1993) High-resolution cosmid and P1 maps spanning the 14-Mb genome of the fission yeast S. pombe. Cell 73:109–120
Mitchell RH (1992) Inositol lipids in cellular signalling mechanisms. Trends Biochem Sci 17:274–276
Moreno S, Klar A, Nurse P (1991) An introduction to molecular genetic analysis of the fission yeast Schizosaccharomyces pombe. Methods Enzymol 194:795–823
Nishizuka Y (1992) Intracellular signalling by hydrolysis of phospholipids and activation of protein kinase C. Science 258:607–614
Payne WE, Fitzgerald-Hayes M (1993) A mutation in PLC1, a candi-date phosphoinositide-specific phospholipase C gene from Saccharomyces cerevisiae, causes aberrant mitotic chromosome segregation. Mol Cell Biol 13:4351–4364
Rhee SG, Choi KD (1992) Multiple forms of phospholipase C isozymes and their activation mechanisms. Adv Second messenger Phosphoprotein Res 26:35–61
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular Cloning. Cold Spring Harbour Laboratory. Cold Spring Harbour, New York
Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chainterminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467
Schweingruber ME, Edenharter E (1990) Thiamin regulates agglutination and zygote formation in Schizosaccharomyces pombe. Curr Genet 17:191–194
Schweingruber AM, Dlugonski J, Edenbarter E, Schweingruber ME (1991) Thiamine in Schizosaccharomyces pombe: dephosphorylation, intracellular pool, biosynthesis and transport. Curr Genet 19:149–254
Yoko-o T, Matsui Y, Yagisawa H, Nojima H, Uno I, Toh-e A (1993) The putative phosphoinositide-specific phospholipase C gene, PLC1, of the yeast Saccharomyces cerevisiae is important for cell growth. Proc Natl Acad Sci USA 90:1804–1808
Zurlinden A, Schweingruber ME (1994) Cloning, nucleotide sequence, and regulation of Schizosaccharomyces pombe thi4, a thiamine biosynthetic gene. J Bacteriol 176:6631–6635
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Communicated by K. Wolf
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Fankhauser, H., Schweingruber, A.M., Edenharter, E. et al. Growth of a mutant defective in a putative phosphoinositide-specific phospholipase C of Schizosaccharomyces pombe is restored by low concentrations of phosphate and inositol. Curr Genet 28, 199–203 (1995). https://doi.org/10.1007/BF00315789
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DOI: https://doi.org/10.1007/BF00315789