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. 2004 May;167(1):77–91. doi: 10.1534/genetics.167.1.77

A novel gene, msa1, inhibits sexual differentiation in Schizosaccharomyces pombe.

Hee Tae Jeong 1, Fumiyo Ozoe 1, Katsunori Tanaka 1, Tsuyoshi Nakagawa 1, Hideyuki Matsuda 1, Makoto Kawamukai 1
PMCID: PMC1470851  PMID: 15166138

Abstract

Sexual differentiation in the fission yeast Schizosaccharomyces pombe is triggered by nutrient starvation or by the presence of mating pheromones. We identified a novel gene, msa1, which encodes a 533-aa putative RNA-binding protein that inhibits sexual differentiation. Disruption of the msa1 gene caused cells to hypersporulate. Intracellular levels of msa1 RNA and Msa1 protein diminished after several hours of nitrogen starvation. Genetic analysis suggested that the function of msa1 is independent of the cAMP pathway and stress-responsive pathway. Deletion of the ras1 gene in diploid cells inhibited sporulation and in haploid cells decreased expression of mating-pheromone-induced genes such as mei2, mam2, ste11, and rep1; simultaneous deletion of msa1 reversed both phenotypes. Overexpression of msa1 decreased activated Ras1(Val17)-induced expression of mam2. Phenotypic hypersporulation was similar between cells with deletion of only rad24 and both msa1 and rad24, but simultaneous deletion of msa1 and msa2/nrd1 additively increased hypersporulation. Therefore, we suggest that the primary function of Msa1 is to negatively regulate sexual differentiation by controlling the expression of Ste11-regulated genes, possibly through the pheromone-signaling pathway.

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Selected References

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  1. Andoh T., Kato T., Jr, Matsui Y., Toh-e A. Phosphoinositide-specific phospholipase C forms a complex with 14-3-3 proteins and is involved in expression of UV resistance in fission yeast. Mol Gen Genet. 1998 Apr;258(1-2):139–147. doi: 10.1007/s004380050716. [DOI] [PubMed] [Google Scholar]
  2. Barr M. M., Tu H., Van Aelst L., Wigler M. Identification of Ste4 as a potential regulator of Byr2 in the sexual response pathway of Schizosaccharomyces pombe. Mol Cell Biol. 1996 Oct;16(10):5597–5603. doi: 10.1128/mcb.16.10.5597. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Chen L., Liu T. H., Walworth N. C. Association of Chk1 with 14-3-3 proteins is stimulated by DNA damage. Genes Dev. 1999 Mar 15;13(6):675–685. doi: 10.1101/gad.13.6.675. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Forsburg S. L., Sherman D. A. General purpose tagging vectors for fission yeast. Gene. 1997 Jun 3;191(2):191–195. doi: 10.1016/s0378-1119(97)00058-9. [DOI] [PubMed] [Google Scholar]
  5. Higuchi Toru, Watanabe Yoshinori, Yamamoto Masayuki. Protein kinase A regulates sexual development and gluconeogenesis through phosphorylation of the Zn finger transcriptional activator Rst2p in fission yeast. Mol Cell Biol. 2002 Jan;22(1):1–11. doi: 10.1128/MCB.22.1.1-11.2002. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Iino Y., Sugimoto A., Yamamoto M. S. pombe pac1+, whose overexpression inhibits sexual development, encodes a ribonuclease III-like RNase. EMBO J. 1991 Jan;10(1):221–226. doi: 10.1002/j.1460-2075.1991.tb07939.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Isshiki T., Mochizuki N., Maeda T., Yamamoto M. Characterization of a fission yeast gene, gpa2, that encodes a G alpha subunit involved in the monitoring of nutrition. Genes Dev. 1992 Dec;6(12B):2455–2462. doi: 10.1101/gad.6.12b.2455. [DOI] [PubMed] [Google Scholar]
  8. Kanoh J., Watanabe Y., Ohsugi M., Iino Y., Yamamoto M. Schizosaccharomyces pombe gad7+ encodes a phosphoprotein with a bZIP domain, which is required for proper G1 arrest and gene expression under nitrogen starvation. Genes Cells. 1996 Apr;1(4):391–408. doi: 10.1046/j.1365-2443.1996.d01-247.x. [DOI] [PubMed] [Google Scholar]
  9. Katayama S., Ozoe F., Kurokawa R., Tanaka K., Nakagawa T., Matsuda H., Kawamukai M. Genetic analysis of the sam mutations, which induce sexual development with no requirement for nutritional starvation in fission yeast. Biosci Biotechnol Biochem. 1996 Jun;60(6):994–999. doi: 10.1271/bbb.60.994. [DOI] [PubMed] [Google Scholar]
  10. Kato T., Jr, Okazaki K., Murakami H., Stettler S., Fantes P. A., Okayama H. Stress signal, mediated by a Hog1-like MAP kinase, controls sexual development in fission yeast. FEBS Lett. 1996 Jan 15;378(3):207–212. doi: 10.1016/0014-5793(95)01442-x. [DOI] [PubMed] [Google Scholar]
  11. Kawamukai M., Ferguson K., Wigler M., Young D. Genetic and biochemical analysis of the adenylyl cyclase of Schizosaccharomyces pombe. Cell Regul. 1991 Feb;2(2):155–164. doi: 10.1091/mbc.2.2.155. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kawamukai M., Gerst J., Field J., Riggs M., Rodgers L., Wigler M., Young D. Genetic and biochemical analysis of the adenylyl cyclase-associated protein, cap, in Schizosaccharomyces pombe. Mol Biol Cell. 1992 Feb;3(2):167–180. doi: 10.1091/mbc.3.2.167. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kawamukai M. Isolation of a novel gene, moc2, encoding a putative RNA helicase as a suppressor of sterile strains in Schizosaccharomyces pombe. Biochim Biophys Acta. 1999 Jul 7;1446(1-2):93–101. doi: 10.1016/s0167-4781(99)00071-8. [DOI] [PubMed] [Google Scholar]
  14. Kitamura K., Katayama S., Dhut S., Sato M., Watanabe Y., Yamamoto M., Toda T. Phosphorylation of Mei2 and Ste11 by Pat1 kinase inhibits sexual differentiation via ubiquitin proteolysis and 14-3-3 protein in fission yeast. Dev Cell. 2001 Sep;1(3):389–399. doi: 10.1016/s1534-5807(01)00037-5. [DOI] [PubMed] [Google Scholar]
  15. Kitamura K., Shimoda C. The Schizosaccharomyces pombe mam2 gene encodes a putative pheromone receptor which has a significant homology with the Saccharomyces cerevisiae Ste2 protein. EMBO J. 1991 Dec;10(12):3743–3751. doi: 10.1002/j.1460-2075.1991.tb04943.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kon N., Schroeder S. C., Krawchuk M. D., Wahls W. P. Regulation of the Mts1-Mts2-dependent ade6-M26 meiotic recombination hot spot and developmental decisions by the Spc1 mitogen-activated protein kinase of fission yeast. Mol Cell Biol. 1998 Dec;18(12):7575–7583. doi: 10.1128/mcb.18.12.7575. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Krawchuk M. D., Wahls W. P. High-efficiency gene targeting in Schizosaccharomyces pombe using a modular, PCR-based approach with long tracts of flanking homology. Yeast. 1999 Sep 30;15(13):1419–1427. doi: 10.1002/(SICI)1097-0061(19990930)15:13<1419::AID-YEA466>3.0.CO;2-Q. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Lopez-Girona A., Furnari B., Mondesert O., Russell P. Nuclear localization of Cdc25 is regulated by DNA damage and a 14-3-3 protein. Nature. 1999 Jan 14;397(6715):172–175. doi: 10.1038/16488. [DOI] [PubMed] [Google Scholar]
  19. Maeda T., Mochizuki N., Yamamoto M. Adenylyl cyclase is dispensable for vegetative cell growth in the fission yeast Schizosaccharomyces pombe. Proc Natl Acad Sci U S A. 1990 Oct;87(20):7814–7818. doi: 10.1073/pnas.87.20.7814. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Maeda T., Watanabe Y., Kunitomo H., Yamamoto M. Cloning of the pka1 gene encoding the catalytic subunit of the cAMP-dependent protein kinase in Schizosaccharomyces pombe. J Biol Chem. 1994 Apr 1;269(13):9632–9637. [PubMed] [Google Scholar]
  21. Maundrell K. Thiamine-repressible expression vectors pREP and pRIP for fission yeast. Gene. 1993 Jan 15;123(1):127–130. doi: 10.1016/0378-1119(93)90551-d. [DOI] [PubMed] [Google Scholar]
  22. Maundrell K. nmt1 of fission yeast. A highly transcribed gene completely repressed by thiamine. J Biol Chem. 1990 Jul 5;265(19):10857–10864. [PubMed] [Google Scholar]
  23. Mochizuki N., Yamamoto M. Reduction in the intracellular cAMP level triggers initiation of sexual development in fission yeast. Mol Gen Genet. 1992 May;233(1-2):17–24. doi: 10.1007/BF00587556. [DOI] [PubMed] [Google Scholar]
  24. Mondesert O., McGowan C. H., Russell P. Cig2, a B-type cyclin, promotes the onset of S in Schizosaccharomyces pombe. Mol Cell Biol. 1996 Apr;16(4):1527–1533. doi: 10.1128/mcb.16.4.1527. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Murata M., Takahashi S., Kagiwada S., Suzuki A., Ohnishi S. pH-dependent membrane fusion and vesiculation of phospholipid large unilamellar vesicles induced by amphiphilic anionic and cationic peptides. Biochemistry. 1992 Feb 25;31(7):1986–1992. doi: 10.1021/bi00122a013. [DOI] [PubMed] [Google Scholar]
  26. Nadin-Davis S. A., Nasim A. A gene which encodes a predicted protein kinase can restore some functions of the ras gene in fission yeast. EMBO J. 1988 Apr;7(4):985–993. doi: 10.1002/j.1460-2075.1988.tb02905.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Neiman A. M., Stevenson B. J., Xu H. P., Sprague G. F., Jr, Herskowitz I., Wigler M., Marcus S. Functional homology of protein kinases required for sexual differentiation in Schizosaccharomyces pombe and Saccharomyces cerevisiae suggests a conserved signal transduction module in eukaryotic organisms. Mol Biol Cell. 1993 Jan;4(1):107–120. doi: 10.1091/mbc.4.1.107. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. 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]
  29. Obara-Ishihara T., Okayama H. A B-type cyclin negatively regulates conjugation via interacting with cell cycle 'start' genes in fission yeast. EMBO J. 1994 Apr 15;13(8):1863–1872. doi: 10.1002/j.1460-2075.1994.tb06455.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Obara T., Nakafuku M., Yamamoto M., Kaziro Y. Isolation and characterization of a gene encoding a G-protein alpha subunit from Schizosaccharomyces pombe: involvement in mating and sporulation pathways. Proc Natl Acad Sci U S A. 1991 Jul 1;88(13):5877–5881. doi: 10.1073/pnas.88.13.5877. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Okazaki N., Okazaki K., Tanaka K., Okayama H. The ste4+ gene, essential for sexual differentiation of Schizosaccharomyces pombe, encodes a protein with a leucine zipper motif. Nucleic Acids Res. 1991 Dec;19(25):7043–7047. doi: 10.1093/nar/19.25.7043. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Ozoe Fumiyo, Kurokawa Rumi, Kobayashi Yasuyo, Jeong Hee Tae, Tanaka Katsunori, Sen Kikuo, Nakagawa Tsuyoshi, Matsuda Hideyuki, Kawamukai Makoto. The 14-3-3 proteins Rad24 and Rad25 negatively regulate Byr2 by affecting its localization in Schizosaccharomyces pombe. Mol Cell Biol. 2002 Oct;22(20):7105–7119. doi: 10.1128/MCB.22.20.7105-7119.2002. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Prentice H. L. High efficiency transformation of Schizosaccharomyces pombe by electroporation. Nucleic Acids Res. 1992 Feb 11;20(3):621–621. doi: 10.1093/nar/20.3.621. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Samejima I., Mackie S., Warbrick E., Weisman R., Fantes P. A. The fission yeast mitotic regulator win1+ encodes an MAP kinase kinase kinase that phosphorylates and activates Wis1 MAP kinase kinase in response to high osmolarity. Mol Biol Cell. 1998 Aug;9(8):2325–2335. doi: 10.1091/mbc.9.8.2325. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Sato Masamitsu, Watanabe Yoshinori, Akiyoshi Yuji, Yamamoto Masayuki. 14-3-3 protein interferes with the binding of RNA to the phosphorylated form of fission yeast meiotic regulator Mei2p. Curr Biol. 2002 Jan 22;12(2):141–145. doi: 10.1016/s0960-9822(01)00654-6. [DOI] [PubMed] [Google Scholar]
  36. Shiozaki K., Russell P. Conjugation, meiosis, and the osmotic stress response are regulated by Spc1 kinase through Atf1 transcription factor in fission yeast. Genes Dev. 1996 Sep 15;10(18):2276–2288. doi: 10.1101/gad.10.18.2276. [DOI] [PubMed] [Google Scholar]
  37. Sugimoto A., Iino Y., Maeda T., Watanabe Y., Yamamoto M. Schizosaccharomyces pombe ste11+ encodes a transcription factor with an HMG motif that is a critical regulator of sexual development. Genes Dev. 1991 Nov;5(11):1990–1999. doi: 10.1101/gad.5.11.1990. [DOI] [PubMed] [Google Scholar]
  38. Sugiyama A., Tanaka K., Okazaki K., Nojima H., Okayama H. A zinc finger protein controls the onset of premeiotic DNA synthesis of fission yeast in a Mei2-independent cascade. EMBO J. 1994 Apr 15;13(8):1881–1887. doi: 10.1002/j.1460-2075.1994.tb06457.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Takahashi K., Murakami S., Chikashige Y., Funabiki H., Niwa O., Yanagida M. A low copy number central sequence with strict symmetry and unusual chromatin structure in fission yeast centromere. Mol Biol Cell. 1992 Jul;3(7):819–835. doi: 10.1091/mbc.3.7.819. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Takeda T., Toda T., Kominami K., Kohnosu A., Yanagida M., Jones N. Schizosaccharomyces pombe atf1+ encodes a transcription factor required for sexual development and entry into stationary phase. EMBO J. 1995 Dec 15;14(24):6193–6208. doi: 10.1002/j.1460-2075.1995.tb00310.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Tanabe Kaori, Ito Noriko, Wakuri Tomomi, Ozoe Fumiyo, Umeda Makoto, Katayama Satoshi, Tanaka Katsunori, Matsuda Hideyuki, Kawamukai Makoto. Sla1, a Schizosaccharomyces pombe homolog of the human La protein, induces ectopic meiosis when its C terminus is truncated. Eukaryot Cell. 2003 Dec;2(6):1274–1287. doi: 10.1128/EC.2.6.1274-1287.2003. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Tanabe Kaori, Tanaka Katsunori, Matsuda Hideyuki, Kawamukai Makoto. Truncated Sla1 induces haploid meiosis through the Pat1-Mei2 system in fission yeast. Biosci Biotechnol Biochem. 2004 Jan;68(1):266–270. doi: 10.1271/bbb.68.266. [DOI] [PubMed] [Google Scholar]
  43. Tanaka K., Nishide J., Okazaki K., Kato H., Niwa O., Nakagawa T., Matsuda H., Kawamukai M., Murakami Y. Characterization of a fission yeast SUMO-1 homologue, pmt3p, required for multiple nuclear events, including the control of telomere length and chromosome segregation. Mol Cell Biol. 1999 Dec;19(12):8660–8672. doi: 10.1128/mcb.19.12.8660. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Toda T., Shimanuki M., Yanagida M. Fission yeast genes that confer resistance to staurosporine encode an AP-1-like transcription factor and a protein kinase related to the mammalian ERK1/MAP2 and budding yeast FUS3 and KSS1 kinases. Genes Dev. 1991 Jan;5(1):60–73. doi: 10.1101/gad.5.1.60. [DOI] [PubMed] [Google Scholar]
  45. Tsukahara K., Yamamoto H., Okayama H. An RNA binding protein negatively controlling differentiation in fission yeast. Mol Cell Biol. 1998 Aug;18(8):4488–4498. doi: 10.1128/mcb.18.8.4488. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Tu H., Barr M., Dong D. L., Wigler M. Multiple regulatory domains on the Byr2 protein kinase. Mol Cell Biol. 1997 Oct;17(10):5876–5887. doi: 10.1128/mcb.17.10.5876. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Wang Y., Xu H. P., Riggs M., Rodgers L., Wigler M. byr2, a Schizosaccharomyces pombe gene encoding a protein kinase capable of partial suppression of the ras1 mutant phenotype. Mol Cell Biol. 1991 Jul;11(7):3554–3563. doi: 10.1128/mcb.11.7.3554. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Watanabe Y., Lino Y., Furuhata K., Shimoda C., Yamamoto M. The S.pombe mei2 gene encoding a crucial molecule for commitment to meiosis is under the regulation of cAMP. EMBO J. 1988 Mar;7(3):761–767. doi: 10.1002/j.1460-2075.1988.tb02873.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Watanabe Y., Shinozaki-Yabana S., Chikashige Y., Hiraoka Y., Yamamoto M. Phosphorylation of RNA-binding protein controls cell cycle switch from mitotic to meiotic in fission yeast. Nature. 1997 Mar 13;386(6621):187–190. doi: 10.1038/386187a0. [DOI] [PubMed] [Google Scholar]
  50. Watanabe Y., Yamamoto M. S. pombe mei2+ encodes an RNA-binding protein essential for premeiotic DNA synthesis and meiosis I, which cooperates with a novel RNA species meiRNA. Cell. 1994 Aug 12;78(3):487–498. doi: 10.1016/0092-8674(94)90426-x. [DOI] [PubMed] [Google Scholar]
  51. Wu S. Y., McLeod M. The sak1+ gene of Schizosaccharomyces pombe encodes an RFX family DNA-binding protein that positively regulates cyclic AMP-dependent protein kinase-mediated exit from the mitotic cell cycle. Mol Cell Biol. 1995 Mar;15(3):1479–1488. doi: 10.1128/mcb.15.3.1479. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Xu H. P., White M., Marcus S., Wigler M. Concerted action of RAS and G proteins in the sexual response pathways of Schizosaccharomyces pombe. Mol Cell Biol. 1994 Jan;14(1):50–58. doi: 10.1128/mcb.14.1.50. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Zhou G. L., Yamamoto T., Ozoe F., Yano D., Tanaka K., Matsuda H., Kawamukai M. Identification of a 14-3-3 protein from Lentinus edodes that interacts with CAP (adenylyl cyclase-associated protein), and conservation of this interaction in fission yeast. Biosci Biotechnol Biochem. 2000 Jan;64(1):149–159. doi: 10.1271/bbb.64.149. [DOI] [PubMed] [Google Scholar]

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