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. 1997 Oct 15;16(20):6171–6181. doi: 10.1093/emboj/16.20.6171

Yeast spore germination: a requirement for Ras protein activity during re-entry into the cell cycle.

P K Herman 1, J Rine 1
PMCID: PMC1326301  PMID: 9321396

Abstract

Saccharomyces cerevisiae spore germination is a process in which quiescent, non-dividing spores become competent for mitotic cell division. Using a novel assay for spore uncoating, we found that spore germination was a multi-step process whose nutritional requirements differed from those for mitotic division. Although both processes were controlled by nutrient availability, efficient spore germination occurred in conditions that did not support cell division. In addition, germination did not require many key regulators of cell cycle progression including the cyclin-dependent kinase, Cdc28p. However, two processes essential for cell growth, protein synthesis and signaling through the Ras protein pathway, were required for spore germination. Moreover, increasing Ras protein activity in spores resulted in an accelerated rate of germination and suggested that activation of the Ras pathway was rate-limiting for entry into the germination program. An early step in germination, commitment, was identified as the point at which spores became irreversibly destined to complete the uncoating process even if the original stimulus for germination was removed. Spore commitment to germination required protein synthesis and Ras protein activity; in contrast, post-commitment events did not require ongoing protein synthesis. Altogether, these data suggested a model for Ras function during transitions between periods of quiescence and cell cycle progression.

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Selected References

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  1. Alani E., Cao L., Kleckner N. A method for gene disruption that allows repeated use of URA3 selection in the construction of multiply disrupted yeast strains. Genetics. 1987 Aug;116(4):541–545. doi: 10.1534/genetics.112.541.test. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Ballou C. E., Maitra S. K., Walker J. W., Whelan W. L. Developmental defects associated with glucosamine auxotrophy in Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1977 Oct;74(10):4351–4355. doi: 10.1073/pnas.74.10.4351. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Barnes D. A., Thorner J. Genetic manipulation of Saccharomyces cerevisiae by use of the LYS2 gene. Mol Cell Biol. 1986 Aug;6(8):2828–2838. doi: 10.1128/mcb.6.8.2828. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Brenner C., Nakayama N., Goebl M., Tanaka K., Toh-e A., Matsumoto K. CDC33 encodes mRNA cap-binding protein eIF-4E of Saccharomyces cerevisiae. Mol Cell Biol. 1988 Aug;8(8):3556–3559. doi: 10.1128/mcb.8.8.3556. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Briza P., Ellinger A., Winkler G., Breitenbach M. Characterization of a DL-dityrosine-containing macromolecule from yeast ascospore walls. J Biol Chem. 1990 Sep 5;265(25):15118–15123. [PubMed] [Google Scholar]
  6. Briza P., Ellinger A., Winkler G., Breitenbach M. Chemical composition of the yeast ascospore wall. The second outer layer consists of chitosan. J Biol Chem. 1988 Aug 15;263(23):11569–11574. [PubMed] [Google Scholar]
  7. Briza P., Winkler G., Kalchhauser H., Breitenbach M. Dityrosine is a prominent component of the yeast ascospore wall. A proof of its structure. J Biol Chem. 1986 Mar 25;261(9):4288–4294. [PubMed] [Google Scholar]
  8. Broach J. R. RAS genes in Saccharomyces cerevisiae: signal transduction in search of a pathway. Trends Genet. 1991 Jan;7(1):28–33. doi: 10.1016/0168-9525(91)90018-l. [DOI] [PubMed] [Google Scholar]
  9. Broek D., Toda T., Michaeli T., Levin L., Birchmeier C., Zoller M., Powers S., Wigler M. The S. cerevisiae CDC25 gene product regulates the RAS/adenylate cyclase pathway. Cell. 1987 Mar 13;48(5):789–799. doi: 10.1016/0092-8674(87)90076-6. [DOI] [PubMed] [Google Scholar]
  10. Choih S. J., Ferro A. J., Shapiro S. K. Function of S-adenosylmethionine in germinating yeast ascospores. J Bacteriol. 1977 Jul;131(1):63–68. doi: 10.1128/jb.131.1.63-68.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Danaie P., Wittmer B., Altmann M., Trachsel H. Isolation of a protein complex containing translation initiation factor Prt1 from Saccharomyces cerevisiae. J Biol Chem. 1995 Mar 3;270(9):4288–4292. doi: 10.1074/jbc.270.9.4288. [DOI] [PubMed] [Google Scholar]
  12. Dawes I. W., Hardie I. D. Selective killing of vegetative cells in sporulated yeast cultures by exposure to diethyl ether. Mol Gen Genet. 1974;131(4):281–289. doi: 10.1007/BF00264859. [DOI] [PubMed] [Google Scholar]
  13. Donnini C., Artoni N., Marmiroli N. Germination conditions that require mitochondrial function in Saccharomyces cerevisiae: utilization of acetate and galactose. J Bacteriol. 1986 Dec;168(3):1250–1253. doi: 10.1128/jb.168.3.1250-1253.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Gibbs J. B., Marshall M. S. The ras oncogene--an important regulatory element in lower eucaryotic organisms. Microbiol Rev. 1989 Jun;53(2):171–185. doi: 10.1128/mr.53.2.171-185.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Granot D., Snyder M. Carbon source induces growth of stationary phase yeast cells, independent of carbon source metabolism. Yeast. 1993 May;9(5):465–479. doi: 10.1002/yea.320090503. [DOI] [PubMed] [Google Scholar]
  16. Granot D., Snyder M. Glucose induces cAMP-independent growth-related changes in stationary-phase cells of Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1991 Jul 1;88(13):5724–5728. doi: 10.1073/pnas.88.13.5724. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. HASHIMOTO T., CONTI S. F., NAYLOR H. B. Fine structure of microorganisms. III. Electron microscopy of resting and germinating ascospores of Saccharomyces cerevisiae. J Bacteriol. 1958 Oct;76(4):406–416. doi: 10.1128/jb.76.4.406-416.1958. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hubler L., Bradshaw-Rouse J., Heideman W. Connections between the Ras-cyclic AMP pathway and G1 cyclin expression in the budding yeast Saccharomyces cerevisiae. Mol Cell Biol. 1993 Oct;13(10):6274–6282. doi: 10.1128/mcb.13.10.6274. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Ito H., Fukuda Y., Murata K., Kimura A. Transformation of intact yeast cells treated with alkali cations. J Bacteriol. 1983 Jan;153(1):163–168. doi: 10.1128/jb.153.1.163-168.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kreger-Van Rij N. J. Electron microscopy of germinating ascospores of Saccharomyces cerevisiae. Arch Microbiol. 1978 Apr 27;117(1):73–77. doi: 10.1007/BF00689354. [DOI] [PubMed] [Google Scholar]
  21. Matsumoto K., Uno I., Ishikawa T. Control of cell division in Saccharomyces cerevisiae mutants defective in adenylate cyclase and cAMP-dependent protein kinase. Exp Cell Res. 1983 Jun;146(1):151–161. doi: 10.1016/0014-4827(83)90333-6. [DOI] [PubMed] [Google Scholar]
  22. Matsumoto K., Uno I., Oshima Y., Ishikawa T. Isolation and characterization of yeast mutants deficient in adenylate cyclase and cAMP-dependent protein kinase. Proc Natl Acad Sci U S A. 1982 Apr;79(7):2355–2359. doi: 10.1073/pnas.79.7.2355. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Mitchell A. P. Control of meiotic gene expression in Saccharomyces cerevisiae. Microbiol Rev. 1994 Mar;58(1):56–70. doi: 10.1128/mr.58.1.56-70.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Naranda T., MacMillan S. E., Hershey J. W. Purified yeast translational initiation factor eIF-3 is an RNA-binding protein complex that contains the PRT1 protein. J Biol Chem. 1994 Dec 23;269(51):32286–32292. [PubMed] [Google Scholar]
  25. Norbury C., Nurse P. Animal cell cycles and their control. Annu Rev Biochem. 1992;61:441–470. doi: 10.1146/annurev.bi.61.070192.002301. [DOI] [PubMed] [Google Scholar]
  26. Pardee A. B. G1 events and regulation of cell proliferation. Science. 1989 Nov 3;246(4930):603–608. doi: 10.1126/science.2683075. [DOI] [PubMed] [Google Scholar]
  27. Pausch M. H., Kaim D., Kunisawa R., Admon A., Thorner J. Multiple Ca2+/calmodulin-dependent protein kinase genes in a unicellular eukaryote. EMBO J. 1991 Jun;10(6):1511–1522. doi: 10.1002/j.1460-2075.1991.tb07671.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Reed S. I. The role of p34 kinases in the G1 to S-phase transition. Annu Rev Cell Biol. 1992;8:529–561. doi: 10.1146/annurev.cb.08.110192.002525. [DOI] [PubMed] [Google Scholar]
  29. Rousseau P., Halvorson H. O. Effect of metabolic inhibitors on germination and outgrowth of Saccharomyces cerevisiae ascospores. Can J Microbiol. 1973 Oct;19(10):1311–1318. doi: 10.1139/m73-210. [DOI] [PubMed] [Google Scholar]
  30. Rousseau P., Halvorson H. O. Preparation and storage of single spores of Saccharomyces cerevisiae. J Bacteriol. 1969 Dec;100(3):1426–1427. doi: 10.1128/jb.100.3.1426-1427.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Savarese J. J. Germination studies on pure yeast ascospores. Can J Microbiol. 1974 Nov;20(11):1517–1522. doi: 10.1139/m74-237. [DOI] [PubMed] [Google Scholar]
  32. Schiestl R. H., Gietz R. D. High efficiency transformation of intact yeast cells using single stranded nucleic acids as a carrier. Curr Genet. 1989 Dec;16(5-6):339–346. doi: 10.1007/BF00340712. [DOI] [PubMed] [Google Scholar]
  33. Seigel J. L., Miller J. J. Observations on acid-fastness and respiration of germinating yeast ascospores. Can J Microbiol. 1971 Jul;17(7):837–845. doi: 10.1139/m71-135. [DOI] [PubMed] [Google Scholar]
  34. Seufert W., McGrath J. P., Jentsch S. UBC1 encodes a novel member of an essential subfamily of yeast ubiquitin-conjugating enzymes involved in protein degradation. EMBO J. 1990 Dec;9(13):4535–4541. doi: 10.1002/j.1460-2075.1990.tb07905.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Stotz A., Linder P. The ADE2 gene from Saccharomyces cerevisiae: sequence and new vectors. Gene. 1990 Oct 30;95(1):91–98. doi: 10.1016/0378-1119(90)90418-q. [DOI] [PubMed] [Google Scholar]
  36. Thevelein J. M. Signal transduction in yeast. Yeast. 1994 Dec;10(13):1753–1790. doi: 10.1002/yea.320101308. [DOI] [PubMed] [Google Scholar]
  37. Tingle M. A., Küenzi M. T., Halvorson H. O. Germination of yeast spores lacking mitochondrial deoxyribonucleic acid. J Bacteriol. 1974 Jan;117(1):89–93. doi: 10.1128/jb.117.1.89-93.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Toda T., Uno I., Ishikawa T., Powers S., Kataoka T., Broek D., Cameron S., Broach J., Matsumoto K., Wigler M. In yeast, RAS proteins are controlling elements of adenylate cyclase. Cell. 1985 Jan;40(1):27–36. doi: 10.1016/0092-8674(85)90305-8. [DOI] [PubMed] [Google Scholar]
  39. Werner-Washburne M., Braun E., Johnston G. C., Singer R. A. Stationary phase in the yeast Saccharomyces cerevisiae. Microbiol Rev. 1993 Jun;57(2):383–401. doi: 10.1128/mr.57.2.383-401.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Xu G., West T. P. Nutritional and physiological factors affecting germination of heterothallic Saccharomyces cerevisiae ascospores. Microbios. 1992;72(290):27–34. [PubMed] [Google Scholar]

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