Skip to main content
NCBI home page
The EMBO Journal logoLink to The EMBO Journal
. 1996 Jul 15;15(14):3599–3610.

Two S-phase checkpoint systems, one involving the function of both BIME and Tyr15 phosphorylation of p34cdc2, inhibit NIMA and prevent premature mitosis.

X S Ye 1, R R Fincher 1, A Tang 1, K O'Donnell 1, S A Osmani 1
PMCID: PMC451970  PMID: 8670863

Abstract

We demonstrate that there are at least two S-phase checkpoint mechanisms controlling mitosis in Aspergillus. The first responds to the rate of DNA replication and inhibits mitosis via tyrosine phosphorylation of p34cdc2. Cells unable to tyrosine phosphorylate p34cdc2 are therefore viable but are unable to tolerate low levels of hydroxyurea and prematurely enter lethal mitosis when S-phase is slowed. However, if the NIMA mitosis-promoting kinase is inactivated then non-tyrosine-phosphorylated p34cdc2 cannot promote cells prematurely into mitosis. Lack of tyrosine-phosphorylated p34cdc2 also cannot promote mitosis, or lethality, if DNA replication is arrested, demonstrating the presence of a second S-phase checkpoint mechanism over mitotic initiation which we show involves the function of BIME. In order to overcome the S-phase arrest checkpoint over mitosis it is necessary both to prevent tyrosine phosphorylation of p34cdc2 and also to inactivate BIME. Lack of tyrosine phosphorylation of p34cdc2 allows precocious expression of NIMA during S-phase arrest, and lack of BIME then allows activation of this prematurely expressed NIMA by phosphorylation. The mitosis-promoting NIMA kinase is thus a target for S-phase checkpoint controls.

Full text

PDF
3599

Images in this article

3601Image
on p.3601
3602Image
on p.3602
3603Image
on p.3603
3605Image
on p.3605
3606Image
on p.3606

Selected References

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

  1. Amon A., Surana U., Muroff I., Nasmyth K. Regulation of p34CDC28 tyrosine phosphorylation is not required for entry into mitosis in S. cerevisiae. Nature. 1992 Jan 23;355(6358):368–371. doi: 10.1038/355368a0. [DOI] [PubMed] [Google Scholar]
  2. Bergen L. G., Morris N. R. Kinetics of the nuclear division cycle of Aspergillus nidulans. J Bacteriol. 1983 Oct;156(1):155–160. doi: 10.1128/jb.156.1.155-160.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bergen L. G., Upshall A., Morris N. R. S-phase, G2, and nuclear division mutants of Aspergillus nidulans. J Bacteriol. 1984 Jul;159(1):114–119. doi: 10.1128/jb.159.1.114-119.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Booher R. N., Alfa C. E., Hyams J. S., Beach D. H. The fission yeast cdc2/cdc13/suc1 protein kinase: regulation of catalytic activity and nuclear localization. Cell. 1989 Aug 11;58(3):485–497. doi: 10.1016/0092-8674(89)90429-7. [DOI] [PubMed] [Google Scholar]
  5. Correa-Bordes J., Nurse P. p25rum1 orders S phase and mitosis by acting as an inhibitor of the p34cdc2 mitotic kinase. Cell. 1995 Dec 15;83(6):1001–1009. doi: 10.1016/0092-8674(95)90215-5. [DOI] [PubMed] [Google Scholar]
  6. Den Haese G. J., Walworth N., Carr A. M., Gould K. L. The Wee1 protein kinase regulates T14 phosphorylation of fission yeast Cdc2. Mol Biol Cell. 1995 Apr;6(4):371–385. doi: 10.1091/mbc.6.4.371. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Doonan J. H., MacKintosh C., Osmani S., Cohen P., Bai G., Lee E. Y., Morris N. R. A cDNA encoding rabbit muscle protein phosphatase 1 alpha complements the Aspergillus cell cycle mutation, bimG11. J Biol Chem. 1991 Oct 5;266(28):18889–18894. [PubMed] [Google Scholar]
  8. Draetta G. Cell cycle control in eukaryotes: molecular mechanisms of cdc2 activation. Trends Biochem Sci. 1990 Oct;15(10):378–383. doi: 10.1016/0968-0004(90)90235-4. [DOI] [PubMed] [Google Scholar]
  9. Dunphy W. G. The decision to enter mitosis. Trends Cell Biol. 1994 Jun;4(6):202–207. doi: 10.1016/0962-8924(94)90142-2. [DOI] [PubMed] [Google Scholar]
  10. Edgar B. A., O'Farrell P. H. The three postblastoderm cell cycles of Drosophila embryogenesis are regulated in G2 by string. Cell. 1990 Aug 10;62(3):469–480. doi: 10.1016/0092-8674(90)90012-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Enoch T., Carr A. M., Nurse P. Fission yeast genes involved in coupling mitosis to completion of DNA replication. Genes Dev. 1992 Nov;6(11):2035–2046. doi: 10.1101/gad.6.11.2035. [DOI] [PubMed] [Google Scholar]
  12. Enoch T., Nurse P. Mutation of fission yeast cell cycle control genes abolishes dependence of mitosis on DNA replication. Cell. 1990 Feb 23;60(4):665–673. doi: 10.1016/0092-8674(90)90669-6. [DOI] [PubMed] [Google Scholar]
  13. Evans T., Rosenthal E. T., Youngblom J., Distel D., Hunt T. Cyclin: a protein specified by maternal mRNA in sea urchin eggs that is destroyed at each cleavage division. Cell. 1983 Jun;33(2):389–396. doi: 10.1016/0092-8674(83)90420-8. [DOI] [PubMed] [Google Scholar]
  14. Fesquet D., Labbé J. C., Derancourt J., Capony J. P., Galas S., Girard F., Lorca T., Shuttleworth J., Dorée M., Cavadore J. C. The MO15 gene encodes the catalytic subunit of a protein kinase that activates cdc2 and other cyclin-dependent kinases (CDKs) through phosphorylation of Thr161 and its homologues. EMBO J. 1993 Aug;12(8):3111–3121. doi: 10.1002/j.1460-2075.1993.tb05980.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Fry A. M., Schultz S. J., Bartek J., Nigg E. A. Substrate specificity and cell cycle regulation of the Nek2 protein kinase, a potential human homolog of the mitotic regulator NIMA of Aspergillus nidulans. J Biol Chem. 1995 May 26;270(21):12899–12905. doi: 10.1074/jbc.270.21.12899. [DOI] [PubMed] [Google Scholar]
  16. Gale M., Jr, Parsons M. A Trypanosoma brucei gene family encoding protein kinases with catalytic domains structurally related to Nek1 and NIMA. Mol Biochem Parasitol. 1993 May;59(1):111–121. doi: 10.1016/0166-6851(93)90012-m. [DOI] [PubMed] [Google Scholar]
  17. Gould K. L., Nurse P. Tyrosine phosphorylation of the fission yeast cdc2+ protein kinase regulates entry into mitosis. Nature. 1989 Nov 2;342(6245):39–45. doi: 10.1038/342039a0. [DOI] [PubMed] [Google Scholar]
  18. Hartwell L. H., Weinert T. A. Checkpoints: controls that ensure the order of cell cycle events. Science. 1989 Nov 3;246(4930):629–634. doi: 10.1126/science.2683079. [DOI] [PubMed] [Google Scholar]
  19. Heald R., McLoughlin M., McKeon F. Human wee1 maintains mitotic timing by protecting the nucleus from cytoplasmically activated Cdc2 kinase. Cell. 1993 Aug 13;74(3):463–474. doi: 10.1016/0092-8674(93)80048-j. [DOI] [PubMed] [Google Scholar]
  20. James S. W., Mirabito P. M., Scacheri P. C., Morris N. R. The Aspergillus nidulans bimE (blocked-in-mitosis) gene encodes multiple cell cycle functions involved in mitotic checkpoint control and mitosis. J Cell Sci. 1995 Nov;108(Pt 11):3485–3499. doi: 10.1242/jcs.108.11.3485. [DOI] [PubMed] [Google Scholar]
  21. Jones D. G., Rosamond J. Isolation of a novel protein kinase-encoding gene from yeast by oligodeoxyribonucleotide probing. Gene. 1990 May 31;90(1):87–92. doi: 10.1016/0378-1119(90)90442-t. [DOI] [PubMed] [Google Scholar]
  22. Krek W., Nigg E. A. Mutations of p34cdc2 phosphorylation sites induce premature mitotic events in HeLa cells: evidence for a double block to p34cdc2 kinase activation in vertebrates. EMBO J. 1991 Nov;10(11):3331–3341. doi: 10.1002/j.1460-2075.1991.tb04897.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kumagai A., Dunphy W. G. Control of the Cdc2/cyclin B complex in Xenopus egg extracts arrested at a G2/M checkpoint with DNA synthesis inhibitors. Mol Biol Cell. 1995 Feb;6(2):199–213. doi: 10.1091/mbc.6.2.199. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Lee T. H., Kirschner M. W. An inhibitor of p34cdc2/cyclin B that regulates the G2/M transition in Xenopus extracts. Proc Natl Acad Sci U S A. 1996 Jan 9;93(1):352–356. doi: 10.1073/pnas.93.1.352. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Letwin K., Mizzen L., Motro B., Ben-David Y., Bernstein A., Pawson T. A mammalian dual specificity protein kinase, Nek1, is related to the NIMA cell cycle regulator and highly expressed in meiotic germ cells. EMBO J. 1992 Oct;11(10):3521–3531. doi: 10.1002/j.1460-2075.1992.tb05435.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Levedakou E. N., He M., Baptist E. W., Craven R. J., Cance W. G., Welcsh P. L., Simmons A., Naylor S. L., Leach R. J., Lewis T. B. Two novel human serine/threonine kinases with homologies to the cell cycle regulating Xenopus MO15, and NIMA kinases: cloning and characterization of their expression pattern. Oncogene. 1994 Jul;9(7):1977–1988. [PubMed] [Google Scholar]
  27. Lu K. P., Hunter T. Evidence for a NIMA-like mitotic pathway in vertebrate cells. Cell. 1995 May 5;81(3):413–424. doi: 10.1016/0092-8674(95)90394-1. [DOI] [PubMed] [Google Scholar]
  28. Lu K. P., Hunter T. The NIMA kinase: a mitotic regulator in Aspergillus nidulans and vertebrate cells. Prog Cell Cycle Res. 1995;1:187–205. doi: 10.1007/978-1-4615-1809-9_15. [DOI] [PubMed] [Google Scholar]
  29. Lu K. P., Means A. R. Expression of the noncatalytic domain of the NIMA kinase causes a G2 arrest in Aspergillus nidulans. EMBO J. 1994 May 1;13(9):2103–2113. doi: 10.1002/j.1460-2075.1994.tb06486.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Lu K. P., Osmani S. A., Means A. R. Properties and regulation of the cell cycle-specific NIMA protein kinase of Aspergillus nidulans. J Biol Chem. 1993 Apr 25;268(12):8769–8776. [PubMed] [Google Scholar]
  31. Lundgren K., Walworth N., Booher R., Dembski M., Kirschner M., Beach D. mik1 and wee1 cooperate in the inhibitory tyrosine phosphorylation of cdc2. Cell. 1991 Mar 22;64(6):1111–1122. doi: 10.1016/0092-8674(91)90266-2. [DOI] [PubMed] [Google Scholar]
  32. Morris N. R. Mitotic mutants of Aspergillus nidulans. Genet Res. 1975 Dec;26(3):237–254. doi: 10.1017/s0016672300016049. [DOI] [PubMed] [Google Scholar]
  33. Mueller P. R., Coleman T. R., Kumagai A., Dunphy W. G. Myt1: a membrane-associated inhibitory kinase that phosphorylates Cdc2 on both threonine-14 and tyrosine-15. Science. 1995 Oct 6;270(5233):86–90. doi: 10.1126/science.270.5233.86. [DOI] [PubMed] [Google Scholar]
  34. Murray A. W. Creative blocks: cell-cycle checkpoints and feedback controls. Nature. 1992 Oct 15;359(6396):599–604. doi: 10.1038/359599a0. [DOI] [PubMed] [Google Scholar]
  35. Murray A. W. The genetics of cell cycle checkpoints. Curr Opin Genet Dev. 1995 Feb;5(1):5–11. doi: 10.1016/s0959-437x(95)90046-2. [DOI] [PubMed] [Google Scholar]
  36. Navas T. A., Zhou Z., Elledge S. J. DNA polymerase epsilon links the DNA replication machinery to the S phase checkpoint. Cell. 1995 Jan 13;80(1):29–39. doi: 10.1016/0092-8674(95)90448-4. [DOI] [PubMed] [Google Scholar]
  37. 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]
  38. O'Connell M. J., Norbury C., Nurse P. Premature chromatin condensation upon accumulation of NIMA. EMBO J. 1994 Oct 17;13(20):4926–4937. doi: 10.1002/j.1460-2075.1994.tb06820.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. O'Dorisio M. S., Chen F., O'Dorisio T. M., Wray D., Qualman S. J. Characterization of somatostatin receptors on human neuroblastoma tumors. Cell Growth Differ. 1994 Jan;5(1):1–8. [PubMed] [Google Scholar]
  40. Oakley B. R., Morris N. R. A mutation in Aspergillus nidulans that blocks the transition from interphase to prophase. J Cell Biol. 1983 Apr;96(4):1155–1158. doi: 10.1083/jcb.96.4.1155. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Osmani A. H., McGuire S. L., Osmani S. A. Parallel activation of the NIMA and p34cdc2 cell cycle-regulated protein kinases is required to initiate mitosis in A. nidulans. Cell. 1991 Oct 18;67(2):283–291. doi: 10.1016/0092-8674(91)90180-7. [DOI] [PubMed] [Google Scholar]
  42. Osmani A. H., O'Donnell K., Pu R. T., Osmani S. A. Activation of the nimA protein kinase plays a unique role during mitosis that cannot be bypassed by absence of the bimE checkpoint. EMBO J. 1991 Sep;10(9):2669–2679. doi: 10.1002/j.1460-2075.1991.tb07810.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Osmani A. H., van Peij N., Mischke M., O'Connell M. J., Osmani S. A. A single p34cdc2 protein kinase (encoded by nimXcdc2) is required at G1 and G2 in Aspergillus nidulans. J Cell Sci. 1994 Jun;107(Pt 6):1519–1528. doi: 10.1242/jcs.107.6.1519. [DOI] [PubMed] [Google Scholar]
  44. Osmani S. A., Engle D. B., Doonan J. H., Morris N. R. Spindle formation and chromatin condensation in cells blocked at interphase by mutation of a negative cell cycle control gene. Cell. 1988 Jan 29;52(2):241–251. doi: 10.1016/0092-8674(88)90513-2. [DOI] [PubMed] [Google Scholar]
  45. Osmani S. A., May G. S., Morris N. R. Regulation of the mRNA levels of nimA, a gene required for the G2-M transition in Aspergillus nidulans. J Cell Biol. 1987 Jun;104(6):1495–1504. doi: 10.1083/jcb.104.6.1495. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Osmani S. A., Pu R. T., Morris N. R. Mitotic induction and maintenance by overexpression of a G2-specific gene that encodes a potential protein kinase. Cell. 1988 Apr 22;53(2):237–244. doi: 10.1016/0092-8674(88)90385-6. [DOI] [PubMed] [Google Scholar]
  47. Paulovich A. G., Hartwell L. H. A checkpoint regulates the rate of progression through S phase in S. cerevisiae in response to DNA damage. Cell. 1995 Sep 8;82(5):841–847. doi: 10.1016/0092-8674(95)90481-6. [DOI] [PubMed] [Google Scholar]
  48. Poon R. Y., Yamashita K., Adamczewski J. P., Hunt T., Shuttleworth J. The cdc2-related protein p40MO15 is the catalytic subunit of a protein kinase that can activate p33cdk2 and p34cdc2. EMBO J. 1993 Aug;12(8):3123–3132. doi: 10.1002/j.1460-2075.1993.tb05981.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Pu R. T., Osmani S. A. Mitotic destruction of the cell cycle regulated NIMA protein kinase of Aspergillus nidulans is required for mitotic exit. EMBO J. 1995 Mar 1;14(5):995–1003. doi: 10.1002/j.1460-2075.1995.tb07080.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Pu R. T., Xu G., Wu L., Vierula J., O'Donnell K., Ye X. S., Osmani S. A. Isolation of a functional homolog of the cell cycle-specific NIMA protein kinase of Aspergillus nidulans and functional analysis of conserved residues. J Biol Chem. 1995 Jul 28;270(30):18110–18116. doi: 10.1074/jbc.270.30.18110. [DOI] [PubMed] [Google Scholar]
  51. Russell P., Nurse P. cdc25+ functions as an inducer in the mitotic control of fission yeast. Cell. 1986 Apr 11;45(1):145–153. doi: 10.1016/0092-8674(86)90546-5. [DOI] [PubMed] [Google Scholar]
  52. Schultz S. J., Nigg E. A. Identification of 21 novel human protein kinases, including 3 members of a family related to the cell cycle regulator nimA of Aspergillus nidulans. Cell Growth Differ. 1993 Oct;4(10):821–830. [PubMed] [Google Scholar]
  53. Solomon M. J., Harper J. W., Shuttleworth J. CAK, the p34cdc2 activating kinase, contains a protein identical or closely related to p40MO15. EMBO J. 1993 Aug;12(8):3133–3142. doi: 10.1002/j.1460-2075.1993.tb05982.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Sorger P. K., Murray A. W. S-phase feedback control in budding yeast independent of tyrosine phosphorylation of p34cdc28. Nature. 1992 Jan 23;355(6358):365–368. doi: 10.1038/355365a0. [DOI] [PubMed] [Google Scholar]
  55. Starborg M., Brundell E., Gell K., Hög C. A novel murine gene encoding a 216-kDa protein is related to a mitotic checkpoint regulator previously identified in Aspergillus nidulans. J Biol Chem. 1994 Sep 30;269(39):24133–24137. [PubMed] [Google Scholar]
  56. Stueland C. S., Lew D. J., Cismowski M. J., Reed S. I. Full activation of p34CDC28 histone H1 kinase activity is unable to promote entry into mitosis in checkpoint-arrested cells of the yeast Saccharomyces cerevisiae. Mol Cell Biol. 1993 Jun;13(6):3744–3755. doi: 10.1128/mcb.13.6.3744. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. Waring R. B., May G. S., Morris N. R. Characterization of an inducible expression system in Aspergillus nidulans using alcA and tubulin-coding genes. Gene. 1989 Jun 30;79(1):119–130. doi: 10.1016/0378-1119(89)90097-8. [DOI] [PubMed] [Google Scholar]
  58. Weinert T. A., Hartwell L. H. The RAD9 gene controls the cell cycle response to DNA damage in Saccharomyces cerevisiae. Science. 1988 Jul 15;241(4863):317–322. doi: 10.1126/science.3291120. [DOI] [PubMed] [Google Scholar]
  59. Weinert T. A., Kiser G. L., Hartwell L. H. Mitotic checkpoint genes in budding yeast and the dependence of mitosis on DNA replication and repair. Genes Dev. 1994 Mar 15;8(6):652–665. doi: 10.1101/gad.8.6.652. [DOI] [PubMed] [Google Scholar]
  60. Ye X. S., Xu G., Pu R. T., Fincher R. R., McGuire S. L., Osmani A. H., Osmani S. A. The NIMA protein kinase is hyperphosphorylated and activated downstream of p34cdc2/cyclin B: coordination of two mitosis promoting kinases. EMBO J. 1995 Mar 1;14(5):986–994. doi: 10.1002/j.1460-2075.1995.tb07079.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  61. al-Khodairy F., Carr A. M. DNA repair mutants defining G2 checkpoint pathways in Schizosaccharomyces pombe. EMBO J. 1992 Apr;11(4):1343–1350. doi: 10.1002/j.1460-2075.1992.tb05179.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES