Abstract
Conjugation in the fission yeast Schizosaccharomyces pombe is controlled by the action of mating pheromones. Here I describe the isolation and characterization of M-factor, the pheromone released by M-type cells. M-factor is a nanopeptide in which the carboxy-terminal cysteine residue is carboxy-methylated and S-alkylated, probably with a farnesyl residue: Tyr-Thr-Pro-Lys-Val-Pro-Tyr-Met-Cys(S-farnesyl)-OCH3. Evidence for this structure was obtained by amino acid analysis, mass spectrometry and tandem mass spectrometry of the native M-factor. Two genes encoding the M-factor were also identified and characterized. It appears that M-factor is synthesized as a larger precursor which is post-translationally cleaved and modified to yield the active pheromone. The proposed modifications are consistent with mechanisms known to exist in other yeast and higher eukaryotes.
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Selected References
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- Anderegg R. J., Betz R., Carr S. A., Crabb J. W., Duntze W. Structure of Saccharomyces cerevisiae mating hormone a-factor. Identification of S-farnesyl cysteine as a structural component. J Biol Chem. 1988 Dec 5;263(34):18236–18240. [PubMed] [Google Scholar]
- Becker J. M., Marcus S., Kundu B., Shenbagamurthi P., Naider F. Towards determination of the structure of the Saccharomyces cerevisiae a-factor: an acylated pentadecapeptide blocks a-factor activity. Mol Cell Biol. 1987 Nov;7(11):4122–4124. doi: 10.1128/mcb.7.11.4122. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Betz R., Crabb J. W., Meyer H. E., Wittig R., Duntze W. Amino acid sequences of a-factor mating peptides from Saccharomyces cerevisiae. J Biol Chem. 1987 Jan 15;262(2):546–548. [PubMed] [Google Scholar]
- Chappell T. G., Warren G. A galactosyltransferase from the fission yeast Schizosaccharomyces pombe. J Cell Biol. 1989 Dec;109(6 Pt 1):2693–2702. doi: 10.1083/jcb.109.6.2693. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cross F., Hartwell L. H., Jackson C., Konopka J. B. Conjugation in Saccharomyces cerevisiae. Annu Rev Cell Biol. 1988;4:429–457. doi: 10.1146/annurev.cb.04.110188.002241. [DOI] [PubMed] [Google Scholar]
- Fehrenbacher G., Perry K., Thorner J. Cell-cell recognition in Saccharomyces cerevisiae: regulation of mating-specific adhesion. J Bacteriol. 1978 Jun;134(3):893–901. doi: 10.1128/jb.134.3.893-901.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Finegold A. A., Schafer W. R., Rine J., Whiteway M., Tamanoi F. Common modifications of trimeric G proteins and ras protein: involvement of polyisoprenylation. Science. 1990 Jul 13;249(4965):165–169. doi: 10.1126/science.1695391. [DOI] [PubMed] [Google Scholar]
- Fukui Y., Kaziro Y., Yamamoto M. Mating pheromone-like diffusible factor released by Schizosaccharomyces pombe. EMBO J. 1986 Aug;5(8):1991–1993. doi: 10.1002/j.1460-2075.1986.tb04454.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Glomset J. A., Gelb M. H., Farnsworth C. C. Prenyl proteins in eukaryotic cells: a new type of membrane anchor. Trends Biochem Sci. 1990 Apr;15(4):139–142. doi: 10.1016/0968-0004(90)90213-u. [DOI] [PubMed] [Google Scholar]
- Goodman L. E., Judd S. R., Farnsworth C. C., Powers S., Gelb M. H., Glomset J. A., Tamanoi F. Mutants of Saccharomyces cerevisiae defective in the farnesylation of Ras proteins. Proc Natl Acad Sci U S A. 1990 Dec;87(24):9665–9669. doi: 10.1073/pnas.87.24.9665. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hartwell L. H. Synchronization of haploid yeast cell cycles, a prelude to conjugation. Exp Cell Res. 1973 Jan;76(1):111–117. doi: 10.1016/0014-4827(73)90425-4. [DOI] [PubMed] [Google Scholar]
- Hrycyna C. A., Clarke S. Farnesyl cysteine C-terminal methyltransferase activity is dependent upon the STE14 gene product in Saccharomyces cerevisiae. Mol Cell Biol. 1990 Oct;10(10):5071–5076. doi: 10.1128/mcb.10.10.5071. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hrycyna C. A., Sapperstein S. K., Clarke S., Michaelis S. The Saccharomyces cerevisiae STE14 gene encodes a methyltransferase that mediates C-terminal methylation of a-factor and RAS proteins. EMBO J. 1991 Jul;10(7):1699–1709. doi: 10.1002/j.1460-2075.1991.tb07694.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kamiya Y., Sakurai A., Tamura S., Takahashi N. Structure of rhodotorucine A, a novel lipopeptide, inducing mating tube formation in Rhodosporidium toruloides. Biochem Biophys Res Commun. 1978 Aug 14;83(3):1077–1083. doi: 10.1016/0006-291x(78)91505-x. [DOI] [PubMed] [Google Scholar]
- Kelly M., Burke J., Smith M., Klar A., Beach D. Four mating-type genes control sexual differentiation in the fission yeast. EMBO J. 1988 May;7(5):1537–1547. doi: 10.1002/j.1460-2075.1988.tb02973.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kuchler K., Sterne R. E., Thorner J. Saccharomyces cerevisiae STE6 gene product: a novel pathway for protein export in eukaryotic cells. EMBO J. 1989 Dec 20;8(13):3973–3984. doi: 10.1002/j.1460-2075.1989.tb08580.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kuchler K., Thorner J. Membrane translocation of proteins without hydrophobic signal peptides. Curr Opin Cell Biol. 1990 Aug;2(4):617–624. doi: 10.1016/0955-0674(90)90102-k. [DOI] [PubMed] [Google Scholar]
- Kurjan J., Herskowitz I. Structure of a yeast pheromone gene (MF alpha): a putative alpha-factor precursor contains four tandem copies of mature alpha-factor. Cell. 1982 Oct;30(3):933–943. doi: 10.1016/0092-8674(82)90298-7. [DOI] [PubMed] [Google Scholar]
- Lee M. G., Nurse P. Complementation used to clone a human homologue of the fission yeast cell cycle control gene cdc2. Nature. 1987 May 7;327(6117):31–35. doi: 10.1038/327031a0. [DOI] [PubMed] [Google Scholar]
- Lipke P. N., Taylor A., Ballou C. E. Morphogenic effects of alpha-factor on Saccharomyces cerevisiae a cells. J Bacteriol. 1976 Jul;127(1):610–618. doi: 10.1128/jb.127.1.610-618.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Marcus S., Caldwell G. A., Miller D., Xue C. B., Naider F., Becker J. M. Significance of C-terminal cysteine modifications to the biological activity of the Saccharomyces cerevisiae a-factor mating pheromone. Mol Cell Biol. 1991 Jul;11(7):3603–3612. doi: 10.1128/mcb.11.7.3603. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Marr R. S., Blair L. C., Thorner J. Saccharomyces cerevisiae STE14 gene is required for COOH-terminal methylation of a-factor mating pheromone. J Biol Chem. 1990 Nov 25;265(33):20057–20060. [PubMed] [Google Scholar]
- McGrath J. P., Varshavsky A. The yeast STE6 gene encodes a homologue of the mammalian multidrug resistance P-glycoprotein. Nature. 1989 Aug 3;340(6232):400–404. doi: 10.1038/340400a0. [DOI] [PubMed] [Google Scholar]
- Moreno S., Klar A., Nurse P. Molecular genetic analysis of fission yeast Schizosaccharomyces pombe. Methods Enzymol. 1991;194:795–823. doi: 10.1016/0076-6879(91)94059-l. [DOI] [PubMed] [Google Scholar]
- Mount S. M. A catalogue of splice junction sequences. Nucleic Acids Res. 1982 Jan 22;10(2):459–472. doi: 10.1093/nar/10.2.459. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nielsen O., Egel R. The pat1 protein kinase controls transcription of the mating-type genes in fission yeast. EMBO J. 1990 May;9(5):1401–1406. doi: 10.1002/j.1460-2075.1990.tb08255.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Powers S., Michaelis S., Broek D., Santa Anna S., Field J., Herskowitz I., Wigler M. RAM, a gene of yeast required for a functional modification of RAS proteins and for production of mating pheromone a-factor. Cell. 1986 Nov 7;47(3):413–422. doi: 10.1016/0092-8674(86)90598-2. [DOI] [PubMed] [Google Scholar]
- Reiss Y., Goldstein J. L., Seabra M. C., Casey P. J., Brown M. S. Inhibition of purified p21ras farnesyl:protein transferase by Cys-AAX tetrapeptides. Cell. 1990 Jul 13;62(1):81–88. doi: 10.1016/0092-8674(90)90242-7. [DOI] [PubMed] [Google Scholar]
- Sakagami Y., Yoshida M., Isogai A., Suzuki A. Peptidal Sex Hormones Inducing Conjugation Tube Formation in Compatible Mating-Type Cells of Tremella mesenterica. Science. 1981 Jun 26;212(4502):1525–1527. doi: 10.1126/science.212.4502.1525. [DOI] [PubMed] [Google Scholar]
- Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schaber M. D., O'Hara M. B., Garsky V. M., Mosser S. C., Bergstrom J. D., Moores S. L., Marshall M. S., Friedman P. A., Dixon R. A., Gibbs J. B. Polyisoprenylation of Ras in vitro by a farnesyl-protein transferase. J Biol Chem. 1990 Sep 5;265(25):14701–14704. [PubMed] [Google Scholar]
- Schafer W. R., Trueblood C. E., Yang C. C., Mayer M. P., Rosenberg S., Poulter C. D., Kim S. H., Rine J. Enzymatic coupling of cholesterol intermediates to a mating pheromone precursor and to the ras protein. Science. 1990 Sep 7;249(4973):1133–1139. doi: 10.1126/science.2204115. [DOI] [PubMed] [Google Scholar]
- Singh A., Chen E. Y., Lugovoy J. M., Chang C. N., Hitzeman R. A., Seeburg P. H. Saccharomyces cerevisiae contains two discrete genes coding for the alpha-factor pheromone. Nucleic Acids Res. 1983 Jun 25;11(12):4049–4063. doi: 10.1093/nar/11.12.4049. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stephenson R. C., Clarke S. Identification of a C-terminal protein carboxyl methyltransferase in rat liver membranes utilizing a synthetic farnesyl cysteine-containing peptide substrate. J Biol Chem. 1990 Sep 25;265(27):16248–16254. [PubMed] [Google Scholar]
- Stetler G. L., Thorner J. Molecular cloning of hormone-responsive genes from the yeast Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1984 Feb;81(4):1144–1148. doi: 10.1073/pnas.81.4.1144. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tsuchiya E., Fukui S., Kamiya Y., Sakagami Y., Fujino M. Requirements of chemical structure of hormonal activity of lipopeptidyl factors inducing sexual differentiation in vegetative cells of heterobasidiomycetous yeasts. Biochem Biophys Res Commun. 1978 Nov 14;85(1):459–463. doi: 10.1016/s0006-291x(78)80064-3. [DOI] [PubMed] [Google Scholar]
- Xue C. B., Caldwell G. A., Becker J. M., Naider F. Total synthesis of the lipopeptide a-mating factor of Saccharomyces cerevisiae. Biochem Biophys Res Commun. 1989 Jul 14;162(1):253–257. doi: 10.1016/0006-291x(89)91989-x. [DOI] [PubMed] [Google Scholar]