Skip to main content
PMC home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1993 Aug 11;21(16):3615–3621. doi: 10.1093/nar/21.16.3615

Domains of p85cdc10 required for function of the fission yeast DSC-1 factor.

A Reymond 1, V Simanis 1
PMCID: PMC309855  PMID: 8367276

Abstract

p85cdc10 is a component of the S.pombe DSC-1 complex, which is thought to mediate periodic transcription of genes in late G1. In order to understand the role of p85cdc10 in the function of this complex, we have analysed which domains of p85cdc10 are required for biological activity and the formation of a stable DSC-1 complex in vitro, both in cdc10 temperature sensitive and null backgrounds. No DSC-1 activity is found in the absence of p85cdc10 and the activity of the complex is reduced or absent in all cdc10ts mutants tested. Full biological activity and rescue of a cdc10::ura4+ null allele requires the N-terminal domain, the cdc10/SWI6 repeats and the helical C-terminal region. In the absence of p85cdc10, both the C-terminal and cdc10/SWI6 repeat domains are required for DSC-1 activity in vitro. In a cdc10ts background, rescue of DSC-1 activity and complementation of mutants, requires only expression of the C-terminal domain, though the presence of the cdc10/SWI6 motifs enhances its activity. The N-terminal domain, alone, or in combination with the cdc10/SWI6 motifs, does not have biological activity, and does not restore DSC-1 activity. We conclude that both the C-terminal domain of p85cdc10 is critical for formation of the DSC-1 complex and that the cdc10/SWI6 motifs also play a role, perhaps by stabilizing the complex. Our data also suggest that the S.pombe DSC-1 complex contains more than one molecule of p85cdc10.

Full text

PDF
3619

Images in this article

3617Image
on p.3617
3618Image
on p.3618
3618Image
on p.3618
3618Image
on p.3618
3619Image
on p.3619

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Andrews B. J., Herskowitz I. The yeast SWI4 protein contains a motif present in developmental regulators and is part of a complex involved in cell-cycle-dependent transcription. Nature. 1989 Dec 14;342(6251):830–833. doi: 10.1038/342830a0. [DOI] [PubMed] [Google Scholar]
  2. Andrews B. J., Moore L. A. Interaction of the yeast Swi4 and Swi6 cell cycle regulatory proteins in vitro. Proc Natl Acad Sci U S A. 1992 Dec 15;89(24):11852–11856. doi: 10.1073/pnas.89.24.11852. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Aves S. J., Durkacz B. W., Carr A., Nurse P. Cloning, sequencing and transcriptional control of the Schizosaccharomyces pombe cdc10 'start' gene. EMBO J. 1985 Feb;4(2):457–463. doi: 10.1002/j.1460-2075.1985.tb03651.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bartlett R., Nurse P. Yeast as a model system for understanding the control of DNA replication in Eukaryotes. Bioessays. 1990 Oct;12(10):457–463. doi: 10.1002/bies.950121002. [DOI] [PubMed] [Google Scholar]
  5. Blank V., Kourilsky P., Israël A. NF-kappa B and related proteins: Rel/dorsal homologies meet ankyrin-like repeats. Trends Biochem Sci. 1992 Apr;17(4):135–140. doi: 10.1016/0968-0004(92)90321-y. [DOI] [PubMed] [Google Scholar]
  6. Breeden L., Nasmyth K. Similarity between cell-cycle genes of budding yeast and fission yeast and the Notch gene of Drosophila. Nature. 1987 Oct 15;329(6140):651–654. doi: 10.1038/329651a0. [DOI] [PubMed] [Google Scholar]
  7. Caligiuri M., Beach D. Sct1 functions in partnership with Cdc10 in a transcription complex that activates cell cycle START and inhibits differentiation. Cell. 1993 Feb 26;72(4):607–619. doi: 10.1016/0092-8674(93)90079-6. [DOI] [PubMed] [Google Scholar]
  8. Davis L. H., Otto E., Bennett V. Specific 33-residue repeat(s) of erythrocyte ankyrin associate with the anion exchanger. J Biol Chem. 1991 Jun 15;266(17):11163–11169. [PubMed] [Google Scholar]
  9. Dirick L., Moll T., Auer H., Nasmyth K. A central role for SWI6 in modulating cell cycle Start-specific transcription in yeast. Nature. 1992 Jun 11;357(6378):508–513. doi: 10.1038/357508a0. [DOI] [PubMed] [Google Scholar]
  10. Fernandez Sarabia M. J., McInerny C., Harris P., Gordon C., Fantes P. The cell cycle genes cdc22+ and suc22+ of the fission yeast Schizosaccharomyces pombe encode the large and small subunits of ribonucleotide reductase. Mol Gen Genet. 1993 Apr;238(1-2):241–251. doi: 10.1007/BF00279553. [DOI] [PubMed] [Google Scholar]
  11. Forsburg S. L., Nurse P. Cell cycle regulation in the yeasts Saccharomyces cerevisiae and Schizosaccharomyces pombe. Annu Rev Cell Biol. 1991;7:227–256. doi: 10.1146/annurev.cb.07.110191.001303. [DOI] [PubMed] [Google Scholar]
  12. Gordon C. B., Fantes P. A. The cdc22 gene of Schizosaccharomyces pombe encodes a cell cycle-regulated transcript. EMBO J. 1986 Nov;5(11):2981–2985. doi: 10.1002/j.1460-2075.1986.tb04595.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Greenwald I., Seydoux G. Analysis of gain-of-function mutations of the lin-12 gene of Caenorhabditis elegans. Nature. 1990 Jul 12;346(6280):197–199. doi: 10.1038/346197a0. [DOI] [PubMed] [Google Scholar]
  14. Hagan I. M., Hyams J. S. The use of cell division cycle mutants to investigate the control of microtubule distribution in the fission yeast Schizosaccharomyces pombe. J Cell Sci. 1988 Mar;89(Pt 3):343–357. doi: 10.1242/jcs.89.3.343. [DOI] [PubMed] [Google Scholar]
  15. Hartwell L. H., Mortimer R. K., Culotti J., Culotti M. Genetic Control of the Cell Division Cycle in Yeast: V. Genetic Analysis of cdc Mutants. Genetics. 1973 Jun;74(2):267–286. doi: 10.1093/genetics/74.2.267. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Johnston L. H. Cell cycle control of gene expression in yeast. Trends Cell Biol. 1992 Dec;2(12):353–357. doi: 10.1016/0962-8924(92)90041-k. [DOI] [PubMed] [Google Scholar]
  17. Kidd S. Characterization of the Drosophila cactus locus and analysis of interactions between cactus and dorsal proteins. Cell. 1992 Nov 13;71(4):623–635. doi: 10.1016/0092-8674(92)90596-5. [DOI] [PubMed] [Google Scholar]
  18. Kodoyianni V., Maine E. M., Kimble J. Molecular basis of loss-of-function mutations in the glp-1 gene of Caenorhabditis elegans. Mol Biol Cell. 1992 Nov;3(11):1199–1213. doi: 10.1091/mbc.3.11.1199. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Lowndes N. F., Johnson A. L., Breeden L., Johnston L. H. SWI6 protein is required for transcription of the periodically expressed DNA synthesis genes in budding yeast. Nature. 1992 Jun 11;357(6378):505–508. doi: 10.1038/357505a0. [DOI] [PubMed] [Google Scholar]
  20. Lowndes N. F., Johnson A. L., Johnston L. H. Coordination of expression of DNA synthesis genes in budding yeast by a cell-cycle regulated trans factor. Nature. 1991 Mar 21;350(6315):247–250. doi: 10.1038/350247a0. [DOI] [PubMed] [Google Scholar]
  21. Lowndes N. F., McInerny C. J., Johnson A. L., Fantes P. A., Johnston L. H. Control of DNA synthesis genes in fission yeast by the cell-cycle gene cdc10+. Nature. 1992 Jan 30;355(6359):449–453. doi: 10.1038/355449a0. [DOI] [PubMed] [Google Scholar]
  22. Marini N. J., Reed S. I. Direct induction of G1-specific transcripts following reactivation of the Cdc28 kinase in the absence of de novo protein synthesis. Genes Dev. 1992 Apr;6(4):557–567. doi: 10.1101/gad.6.4.557. [DOI] [PubMed] [Google Scholar]
  23. Marks J., Fankhauser C., Reymond A., Simanis V. Cytoskeletal and DNA structure abnormalities result from bypass of requirement for the cdc10 start gene in the fission yeast Schizosaccharomyces pombe. J Cell Sci. 1992 Mar;101(Pt 3):517–528. doi: 10.1242/jcs.101.3.517. [DOI] [PubMed] [Google Scholar]
  24. Matsumoto S., Yanagida M., Nurse P. Histone transcription in cell cycle mutants of fission yeast. EMBO J. 1987 Apr;6(4):1093–1097. doi: 10.1002/j.1460-2075.1987.tb04863.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. 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]
  26. Michaely P., Bennett V. The ANK repeat: a ubiquitous motif involved in macromolecular recognition. Trends Cell Biol. 1992 May;2(5):127–129. doi: 10.1016/0962-8924(92)90084-z. [DOI] [PubMed] [Google Scholar]
  27. 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]
  28. Nasmyth K., Dirick L. The role of SWI4 and SWI6 in the activity of G1 cyclins in yeast. Cell. 1991 Sep 6;66(5):995–1013. doi: 10.1016/0092-8674(91)90444-4. [DOI] [PubMed] [Google Scholar]
  29. Nasmyth K., Nurse P., Fraser R. S. The effect of cell mass on the cell cycle timing and duration of S-phase in fission yeast. J Cell Sci. 1979 Oct;39:215–233. doi: 10.1242/jcs.39.1.215. [DOI] [PubMed] [Google Scholar]
  30. Novak B., Mitchison J. M. The first transition point of the mutant cdc2.33 in the fission yeast Schizosaccharomyces pombe. J Cell Sci. 1989 Dec;94(Pt 4):657–662. doi: 10.1242/jcs.94.4.657. [DOI] [PubMed] [Google Scholar]
  31. Nurse P., Bissett Y. Gene required in G1 for commitment to cell cycle and in G2 for control of mitosis in fission yeast. Nature. 1981 Aug 6;292(5823):558–560. doi: 10.1038/292558a0. [DOI] [PubMed] [Google Scholar]
  32. Nurse P. Genetic control of cell size at cell division in yeast. Nature. 1975 Aug 14;256(5518):547–551. doi: 10.1038/256547a0. [DOI] [PubMed] [Google Scholar]
  33. Nurse P., Thuriaux P. Controls over the timing of DNA replication during the cell cycle of fission yeast. Exp Cell Res. 1977 Jul;107(2):365–375. doi: 10.1016/0014-4827(77)90358-5. [DOI] [PubMed] [Google Scholar]
  34. Nurse P., Thuriaux P., Nasmyth K. Genetic control of the cell division cycle in the fission yeast Schizosaccharomyces pombe. Mol Gen Genet. 1976 Jul 23;146(2):167–178. doi: 10.1007/BF00268085. [DOI] [PubMed] [Google Scholar]
  35. Nurse P. Universal control mechanism regulating onset of M-phase. Nature. 1990 Apr 5;344(6266):503–508. doi: 10.1038/344503a0. [DOI] [PubMed] [Google Scholar]
  36. Ogas J., Andrews B. J., Herskowitz I. Transcriptional activation of CLN1, CLN2, and a putative new G1 cyclin (HCS26) by SWI4, a positive regulator of G1-specific transcription. Cell. 1991 Sep 6;66(5):1015–1026. doi: 10.1016/0092-8674(91)90445-5. [DOI] [PubMed] [Google Scholar]
  37. Okazaki K., Okazaki N., Kume K., Jinno S., Tanaka K., Okayama H. High-frequency transformation method and library transducing vectors for cloning mammalian cDNAs by trans-complementation of Schizosaccharomyces pombe. Nucleic Acids Res. 1990 Nov 25;18(22):6485–6489. doi: 10.1093/nar/18.22.6485. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Piggott J. R., Rai R., Carter B. L. A bifunctional gene product involved in two phases of the yeast cell cycle. Nature. 1982 Jul 22;298(5872):391–393. doi: 10.1038/298391a0. [DOI] [PubMed] [Google Scholar]
  39. Primig M., Sockanathan S., Auer H., Nasmyth K. Anatomy of a transcription factor important for the start of the cell cycle in Saccharomyces cerevisiae. Nature. 1992 Aug 13;358(6387):593–597. doi: 10.1038/358593a0. [DOI] [PubMed] [Google Scholar]
  40. Reed S. I. The selection of S. cerevisiae mutants defective in the start event of cell division. Genetics. 1980 Jul;95(3):561–577. doi: 10.1093/genetics/95.3.561. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Reed S. I., Wittenberg C. Mitotic role for the Cdc28 protein kinase of Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1990 Aug;87(15):5697–5701. doi: 10.1073/pnas.87.15.5697. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Reymond A., Schmidt S., Simanis V. Mutations in the cdc10 start gene of Schizosaccharomyces pombe implicate the region of homology between cdc10 and SWI6 as important for p85cdc10 function. Mol Gen Genet. 1992 Sep;234(3):449–456. doi: 10.1007/BF00538705. [DOI] [PubMed] [Google Scholar]
  43. Sidorova J., Breeden L. Analysis of the SWI4/SWI6 protein complex, which directs G1/S-specific transcription in Saccharomyces cerevisiae. Mol Cell Biol. 1993 Feb;13(2):1069–1077. doi: 10.1128/mcb.13.2.1069. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Simanis V., Nurse P. Characterization of the fission yeast cdc10+ protein that is required for commitment to the cell cycle. J Cell Sci. 1989 Jan;92(Pt 1):51–56. doi: 10.1242/jcs.92.1.51. [DOI] [PubMed] [Google Scholar]
  45. Taba M. R., Muroff I., Lydall D., Tebb G., Nasmyth K. Changes in a SWI4,6-DNA-binding complex occur at the time of HO gene activation in yeast. Genes Dev. 1991 Nov;5(11):2000–2013. doi: 10.1101/gad.5.11.2000. [DOI] [PubMed] [Google Scholar]
  46. Tanaka K., Okazaki K., Okazaki N., Ueda T., Sugiyama A., Nojima H., Okayama H. A new cdc gene required for S phase entry of Schizosaccharomyces pombe encodes a protein similar to the cdc 10+ and SWI4 gene products. EMBO J. 1992 Dec;11(13):4923–4932. doi: 10.1002/j.1460-2075.1992.tb05599.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Thompson C. C., Brown T. A., McKnight S. L. Convergence of Ets- and notch-related structural motifs in a heteromeric DNA binding complex. Science. 1991 Aug 16;253(5021):762–768. doi: 10.1126/science.1876833. [DOI] [PubMed] [Google Scholar]
  48. Weilguny D., Praetorius M., Carr A., Egel R., Nielsen O. New vectors in fission yeast: application for cloning the his2 gene. Gene. 1991 Mar 1;99(1):47–54. doi: 10.1016/0378-1119(91)90032-7. [DOI] [PubMed] [Google Scholar]
  49. Woods A., Sherwin T., Sasse R., MacRae T. H., Baines A. J., Gull K. Definition of individual components within the cytoskeleton of Trypanosoma brucei by a library of monoclonal antibodies. J Cell Sci. 1989 Jul;93(Pt 3):491–500. doi: 10.1242/jcs.93.3.491. [DOI] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES