RRC ID 1836
Author Ye Y, Fujii M, Hirata A, Kawamukai M, Shimoda C, Nakamura T.
Title Geranylgeranyl diphosphate synthase in fission yeast is a heteromer of farnesyl diphosphate synthase (FPS), Fps1, and an FPS-like protein, Spo9, essential for sporulation.
Journal Mol. Biol. Cell
Abstract Both farnesyl diphosphate synthase (FPS) and geranylgeranyl diphosphate synthase (GGPS) are key enzymes in the synthesis of various isoprenoid-containing compounds and proteins. Here, we describe two novel Schizosaccharomyces pombe genes, fps1(+) and spo9(+), whose products are similar to FPS in primary structure, but whose functions differ from one another. Fps1 is essential for vegetative growth, whereas, a spo9 null mutant exhibits temperature-sensitive growth. Expression of fps1(+), but not spo9(+), suppresses the lethality of a Saccharomyces cerevisiae FPS-deficient mutant and also restores ubiquinone synthesis in an Escherichia coli ispA mutant, which lacks FPS activity, indicating that S. pombe Fps1 in fact functions as an FPS. In contrast to a typical FPS gene, no apparent GGPS homologues have been found in the S. pombe genome. Interestingly, although neither fps1(+) nor spo9(+) expression alone in E. coli confers clear GGPS activity, coexpression of both genes induces such activity. Moreover, the GGPS activity is significantly reduced in the spo9 mutant. In addition, the spo9 mutation perturbs the membrane association of a geranylgeranylated protein, but not that of a farnesylated protein. Yeast two-hybrid and coimmunoprecipitation analyses indicate that Fps1 and Spo9 physically interact. Thus, neither Fps1 nor Spo9 alone functions as a GGPS, but the two proteins together form a complex with GGPS activity. Because spo9 was originally identified as a sporulation-deficient mutant, we show here that expansion of the forespore membrane is severely inhibited in spo9Delta cells. Electron microscopy revealed significant accumulation membrane vesicles in spo9Delta cells. We suggest that lack of GGPS activity in a spo9 mutant results in impaired protein prenylation in certain proteins responsible for secretory function, thereby inhibiting forespore membrane formation.
Volume 18(9)
Pages 3568-81
Published 2007-9
DOI 10.1091/mbc.e07-02-0112
PII E07-02-0112
PMID 17596513
PMC PMC1951748
MeSH Amino Acid Sequence Bacterial Proteins / metabolism Cell Membrane / metabolism Escherichia coli Farnesyltranstransferase / chemistry Farnesyltranstransferase / genetics Farnesyltranstransferase / metabolism* Genes, Fungal Genetic Complementation Test Geranyltranstransferase / chemistry Geranyltranstransferase / genetics Geranyltranstransferase / metabolism* Molecular Sequence Data Mutation / genetics Phylogeny Protein Binding Protein Structure, Quaternary Saccharomyces cerevisiae / cytology Saccharomyces cerevisiae / enzymology Saccharomyces cerevisiae Proteins / genetics Saccharomyces cerevisiae Proteins / metabolism Schizosaccharomyces / enzymology* Schizosaccharomyces / genetics Schizosaccharomyces / physiology* Schizosaccharomyces / ultrastructure Schizosaccharomyces pombe Proteins / chemistry Schizosaccharomyces pombe Proteins / genetics Schizosaccharomyces pombe Proteins / metabolism* Spores, Fungal / enzymology* Spores, Fungal / ultrastructure Substrate Specificity Transcription, Genetic
IF 3.905
Times Cited 23