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Current Genetics

, Volume 25, Issue 6, pp 497–503 | Cite as

Meiosis-dependent mRNA splicing of the fission yeast Schizosaccharomyces pombe mes1+ gene

  • Masao Kishida
  • Tsutomu Nagai
  • Yukinobu Nakaseko
  • Chikashi Shimoda
Original Articles

Abstract

The mes1+ gene of the fission yeast Schizosaccharomyces pombe is essential for the second meiotic division. We have cloned a 1.1-kb HindIII fragment containing mes1+ by complementation from an S. pombe genomic library. Sequencing of the genomic and cDNA fragments indicates the existence of one small intron of 75 nucleotides, although both the 5′ (G/GTTAGT) and 3′ (CAG/T) intron-exon junctions deviate from the consensus sequences proposed for S. pombe. The putative translation product of the mature mes1+ mRNA is a 11-kDa protein of 101 amino acids which has no significant homology to any previously-reported proteins. Disruption of mes1 has no effect on cell growth but causes an arrest of meiosis before the second meiotic division. Northern-blot analysis revealed that mes1+ was preferentially transcribed under conditions of nitrogen starvation. When a h90 homothallic strain was shifted to a nitrogen-deficient medium, a pre-mRNA accumulated and then was gradually processed to generate a mature mRNA. This splicing did not occur in either a heterothallic haploid strain or in a homothallic mei2 mutant strain which was defective in the initiation of meiosis. Expression of the first exon alone was not able to suppress the mes1 null allele. These results indicate that mes1+ is required for the completion of meiosis, that splicing is required for the function of the mes1+ gene, and that this splicing requires the function of the mei2+ product.

Key words

Fission yeast Intron Meiotic second division Splicing 

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References

  1. Beach D, Nurse P (1981) High-frequency transformation of the fission yeast Schizosaccharomyces pombe. Nature 290:140–142Google Scholar
  2. Beach D, Piper M, Nurse P (1982) Construction of a Schizosaccharomyces pombe gene bank in a yeast bacteria shuttle vector and its use to isolate genes by complementation. Mol Gen Genet 187:326–329Google Scholar
  3. Botstein D, Falco SC, Stewart SE, Brennan M, Scherer S, Stinchcomb DT, Struhl K, Davis RW (1979) Sterile host yeasts (SHY): a eukaryotic system of biological containment for recombinant DNA experiments. Gene 8:17–24Google Scholar
  4. Bresch C, Müller G, Egel R (1968) Genes involved in meiosis and sporulation of a yeast. Mol Gen Genet 102:301–306Google Scholar
  5. Costello G, Rodgers L, Beach D (1986) Fission yeast enters the stationary-phase G0 state from either mitotic G1 or G2. Curr Genet 11:119–125Google Scholar
  6. Egel R (1989) Mating-type genes, meiosis, and sporulation. In: Nasim A, Young P, Johnson BF (eds) Molecular biology of the fission yeast. Academic Press, San Diego, pp 31–73Google Scholar
  7. Egel R, Egel-Mitani M (1974) Premeiotic DNA synthesis in fission yeast. Exp Cell Res 88:127–134Google Scholar
  8. Engebrecht J, Voelkel-Meiman K, Roeder GS (1991) Meiosis-specific RNA splicing in yeast. Cell 66:1257–1268Google Scholar
  9. Esposito RE, Klapholz S (1981) Meiosis and ascospore development. In: Strathern JN, Jones EW, Broach JR (eds) The molecular biology of the yeast Saccharomyces. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, pp 211–287Google Scholar
  10. Fowell RR (1969) Life cycles in yeasts. In: Rose AH, Harrison JS (eds) The yeasts, vol 1. Academic Press, London, pp 303–383Google Scholar
  11. Green MR (1986) Pre-mRNA splicing. Annu Rev Genet 20:671–708Google Scholar
  12. Gutz H, Heslot H, Leupold U, Loprieno N (1974) Schizosaccharomyces pombe. In: King RC (ed) Handbook of genetics, vol. 1. Plenum Press, New York, pp 395–446Google Scholar
  13. Henikoff S (1984) Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene 28:351–359Google Scholar
  14. Hereford L, Fahrener K, Woolford Jr J, Rosbash M, Kaback DB (1979) Isolation of yeast histone genes H2A and H2B. Cell 18:1261–1271Google Scholar
  15. Iino Y, Yamamoto M (1985a) Mutants of Schizosaccharomyces pombe which sporulate in the haploid state. Mol Gen Genet 198:416–421Google Scholar
  16. Iino Y, Yamamoto M (1985b) Negative control for the initiation of meiosis in Schizosaccharomyces pombe. Proc Natl Acad Sci USA 82:2447–2451Google Scholar
  17. Jensen R, Sprague Jr GF, Herskowitz I (1983) Regulation of yeast mating-type interconversion: feedback control of HO gene expression by the mating-type locus. Proc Natl Acad Sci USA 80:3035–3039Google Scholar
  18. Käufer NF, Simanis V, Nurse P (1985) Fission yeast Schizosaccharomyces pombe correctly excises a mammalian RNA transcript intervening sequence. Nature 318:78–80Google Scholar
  19. Laski FA, Rio DC, Rubin GM (1986) Tissue specificity of Drosophila P-element transposition is regulated at the level of mRNA splicing. Cell 44:7–19Google Scholar
  20. Maeda T, Mochizuki N, Yamamoto M (1990) Adenylyl cyclase is dispensable for vegetative cell growth in the fission yeast Schizosaccharomyces pombe. Proc Natl Acad Sci USA 87:7814–7818Google Scholar
  21. Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual, Cold Spring Harbor Laboratory, Cold Spring Harbor, New YorkGoogle Scholar
  22. McLeod M, Beach D (1986) Homology between the ran1 + gene of fission yeast and protein kinases. EMBO J 5:3665–3671Google Scholar
  23. McLeod M, Stein M, Beach D (1987) The product of the mei3 + gene, expressed under control of the mating-type locus, induces meiosis and sporulation in fission yeast. EMBO J 6:729–736Google Scholar
  24. Moreno S, Klar A, Nurse P (1991) Molecular genetic analysis of fission yeast. Methods Enzymol 194:795–823Google Scholar
  25. Nandabalan K, Price L, Roeder GS (1993) Mutations in U1 snRNA bypass the requirement for a cell-type-specific RNA splicing factor. Cell 73:407–415Google Scholar
  26. Nurse P (1985) Mutants of the fission yeast Schizosaccharomyces pombe which alter the shift between cell proliferation and sporulation. Mol Gen Genet 198:497–502Google Scholar
  27. Okazaki K, Okazaki N, Kume K, Jinno S, Tanaka K, Okayama H (1990) High-frequency transformation method and library transducing vectors for cloning mammalian cDNAs by trans-complementation of Schizosaccharomyces pombe. Nucleic Acids Res 18:6485–6489Google Scholar
  28. Rothstein R (1983) One-step gene disruption in yeast. Methods Enzymol 101:202–211Google Scholar
  29. Russell P (1989) Gene cloning and expression in fission yeast. In: Nasim A, Young P, Johnson BF (eds) Molecular biology of the fission yeast. Academic Press, San Diego, pp 243–271Google Scholar
  30. Rymond BC, Rosbash M (1992) Yeast pre-mRNA splicing. In: Jones EW, Pringle JR, Broach JR (eds) The molecular and cellular biology of the yeast Saccharomyces. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, pp 143–192Google Scholar
  31. Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467Google Scholar
  32. Shimoda C, Uehira M (1985) Cloning of the Schizosaccharomyces pombe mei3 gene essential for the initiation of meiosis. Mol Gen Genet 201:353–356Google Scholar
  33. Shimoda C, Hirata A, Kishida M, Hashida T, Tanaka K (1985) Characterization of meiosis-deficient mutants by electron microscopy and mapping of four essential genes in the fission yeast Schizosaccharomyces pombe. Mol Gen Genet 200:252–257Google Scholar
  34. Shimoda C, Uehira M, Kishida M, Fujioka H, Iino Y, Watanabe Y, Yamamoto M (1987) Cloning and analysis of transcription of the mei2 gene responsible for initiation of meiosis in the fission yeast Schizosaccharomyces pombe. J Bacteriol 269:93–96Google Scholar
  35. Thomas PS (1980) Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci USA 77:5201–5205Google Scholar
  36. Watanabe Y, Iino Y, Furuhata H, Yamamoto M (1988) The S. pombe mei2 gene encoding a crucial molecule for commitment to meiosis is under the regulation of cAMP. EMBO J 7:761–767Google Scholar
  37. Yanisch-Perron C, Vieira J, Messing J (1985) Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene 33:103–119Google Scholar

Copyright information

© Springer-Verlag 1994

Authors and Affiliations

  • Masao Kishida
    • 1
  • Tsutomu Nagai
    • 1
  • Yukinobu Nakaseko
    • 2
  • Chikashi Shimoda
    • 1
  1. 1.Department of Biology, Faculty of ScienceOsaka City University, Sumiyoshi-kuOsakaJapan
  2. 2.Department of Biophysics, Faculty of ScienceKyoto University, Sakyo-kuKyotoJapan

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