Home LiteratureArticle Details
PMID: 11598184 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Regulation of the mitotic exit protein kinases Cdc15 and Dbf2.

Molecular biology of the cell ·Vol. 12 ·No. 10 ·2001-10-00 ·Pages 2961-74

Visintin R, Amon A

Abstract

In budding yeast, the release of the protein phosphatase Cdc14 from its inhibitor Cfi1/Net1 in the nucleolus during anaphase triggers the inactivation of Clb CDKs that leads to exit from mitosis. The mitotic exit pathway controls the association between Cdc14 and Cfi1/Net1. It is comprised of the RAS-like GTP binding protein Tem1, the exchange factor Lte1, the GTPase activating protein complex Bub2-Bfa1/Byr4, and several protein kinases including Cdc15 and Dbf2. Here we investigate the regulation of the protein kinases Dbf2 and Cdc15. We find that Cdc15 is recruited to both spindle pole bodies (SPBs) during anaphase. This recruitment depends on TEM1 but not DBF2 or CDC14 and is inhibited by BUB2. Dbf2 also localizes to SPBs during anaphase, which coincides with activation of Dbf2 kinase activity. Both events depend on the mitotic exit pathway components TEM1 and CDC15. In cells lacking BUB2, Dbf2 localized to SPBs in cell cycle stages other than anaphase and telophase and Dbf2 kinase was prematurely active during metaphase. Our results suggest an order of function of mitotic exit pathway components with respect to SPB localization of Cdc15 and Dbf2 and activation of Dbf2 kinase. BUB2 negatively regulates all 3 events. Loading of Cdc15 on SPBs depends on TEM1, whereas loading of Dbf2 on SPBs and activation of Dbf2 kinase depend on TEM1 and CDC15.

MeSH Terms
Anaphase/physiology Cell Cycle Proteins/drug effects,metabolism Enzyme Activation/physiology Fungal Proteins/metabolism,pharmacology GTP-Binding Proteins/drug effects,metabolism Mitosis/physiology Monomeric GTP-Binding Proteins/metabolism Protein Kinases/metabolism Protein Serine-Threonine Kinases Saccharomyces cerevisiae Proteins Saccharomycetales/metabolism Spindle Apparatus/metabolism,ultrastructure
Chemicals
BUB2 protein, S cerevisiae CDC15 protein Cell Cycle Proteins Fungal Proteins Saccharomyces cerevisiae Proteins TEM1 protein, S cerevisiae Protein Kinases DBF2 protein, S cerevisiae Protein Serine-Threonine Kinases GTP-Binding Proteins Monomeric GTP-Binding Proteins
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Visintin R
Center for Cancer Research, Howard Hughes Medical Institute, Massachusetts Institute of Technology, E17-233, 40 Ames Street, Cambridge MA 02139, USA.
Amon A
References (53)
53 references, click to expand
  1. Sid2p, a spindle pole body kinase that regulates the onset of cytokinesis.
    J Cell Biol. 1999 Aug 23;146(4):777-90 PMID: 10459013
  2. The protein kinases of budding yeast: six score and more.
    Trends Biochem Sci. 1997 Jan;22(1):18-22 PMID: 9020587
  3. CDC20 and CDH1: a family of substrate-specific activators of APC-dependent proteolysis.
    Science. 1997 Oct 17;278(5337):460-3 PMID: 9334304
  4. MOB1, an essential yeast gene required for completion of mitosis and maintenance of ploidy.
    Mol Biol Cell. 1998 Jan;9(1):29-46 PMID: 9436989
  5. Sequential assembly of myosin II, an IQGAP-like protein, and filamentous actin to a ring structure involved in budding yeast cytokinesis.
    J Cell Biol. 1998 Jan 26;140(2):355-66 PMID: 9442111
  6. The Polo-like kinase Cdc5p and the WD-repeat protein Cdc20p/fizzy are regulators and substrates of the anaphase promoting complex in Saccharomyces cerevisiae.
    EMBO J. 1998 Mar 2;17(5):1336-49 PMID: 9482731
  7. DBF2 protein kinase binds to and acts through the cell cycle-regulated MOB1 protein.
    Mol Cell Biol. 1998 Apr;18(4):2100-7 PMID: 9528782
  8. The Polo-related kinase Cdc5 activates and is destroyed by the mitotic cyclin destruction machinery in S. cerevisiae.
    Curr Biol. 1998 Apr 23;8(9):497-507 PMID: 9560342
  9. Direct binding of CDC20 protein family members activates the anaphase-promoting complex in mitosis and G1.
    Mol Cell. 1998 Aug;2(2):163-71 PMID: 9734353
  10. A late mitotic regulatory network controlling cyclin destruction in Saccharomyces cerevisiae.
    Mol Biol Cell. 1998 Oct;9(10):2803-17 PMID: 9763445
  11. The phosphatase Cdc14 triggers mitotic exit by reversal of Cdk-dependent phosphorylation.
    Mol Cell. 1998 Dec;2(6):709-18 PMID: 9885559
  12. Inhibitory phosphorylation of the APC regulator Hct1 is controlled by the kinase Cdc28 and the phosphatase Cdc14.
    Curr Biol. 1999 Mar 11;9(5):227-36 PMID: 10074450
  13. Exit from mitosis is triggered by Tem1-dependent release of the protein phosphatase Cdc14 from nucleolar RENT complex.
    Cell. 1999 Apr 16;97(2):233-44 PMID: 10219244
  14. Net1, a Sir2-associated nucleolar protein required for rDNA silencing and nucleolar integrity.
    Cell. 1999 Apr 16;97(2):245-56 PMID: 10219245
  15. Bifurcation of the mitotic checkpoint pathway in budding yeast.
    Proc Natl Acad Sci U S A. 1999 Apr 27;96(9):4989-94 PMID: 10220406
  16. A Bub2p-dependent spindle checkpoint pathway regulates the Dbf2p kinase in budding yeast.
    EMBO J. 1999 May 4;18(9):2424-34 PMID: 10228157
  17. Cfi1 prevents premature exit from mitosis by anchoring Cdc14 phosphatase in the nucleolus.
    Nature. 1999 Apr 29;398(6730):818-23 PMID: 10235265
  18. Sister chromatid separation and chromosome re-duplication are regulated by different mechanisms in response to spindle damage.
    EMBO J. 1999 May 17;18(10):2707-21 PMID: 10329618
  19. Localization of core spindle pole body (SPB) components during SPB duplication in Saccharomyces cerevisiae.
    J Cell Biol. 1999 May 17;145(4):809-23 PMID: 10330408
  20. Budding yeast Bub2 is localized at spindle pole bodies and activates the mitotic checkpoint via a different pathway from Mad2.
    J Cell Biol. 1999 May 31;145(5):979-91 PMID: 10352016
  21. Pds1 and Esp1 control both anaphase and mitotic exit in normal cells and after DNA damage.
    Genes Dev. 1999 Aug 1;13(15):1936-49 PMID: 10444592
  22. Pds1p of budding yeast has dual roles: inhibition of anaphase initiation and regulation of mitotic exit.
    Genes Dev. 1999 Aug 1;13(15):1950-9 PMID: 10444593
  23. Regulation of APC activity by phosphorylation and regulatory factors.
    J Cell Biol. 1999 Aug 23;146(4):791-800 PMID: 10459014
  24. Whose end is destruction: cell division and the anaphase-promoting complex.
    Genes Dev. 1999 Aug 15;13(16):2039-58 PMID: 10465783
  25. Regulation of the APC and the exit from mitosis.
    Nat Cell Biol. 1999 Jun;1(2):E47-53 PMID: 10559897
  26. Progression into and out of mitosis.
    Curr Opin Cell Biol. 1999 Dec;11(6):708-16 PMID: 10600711
  27. APC(Cdc20) promotes exit from mitosis by destroying the anaphase inhibitor Pds1 and cyclin Clb5.
    Nature. 1999 Nov 11;402(6758):203-7 PMID: 10647015
  28. Phosphorylation and spindle pole body localization of the Cdc15p mitotic regulatory protein kinase in budding yeast.
    Curr Biol. 2000 Mar 23;10(6):329-32 PMID: 10744974
  29. Tying the knot: linking cytokinesis to the nuclear cycle.
    J Cell Sci. 2000 May;113 ( Pt 9):1503-13 PMID: 10751142
  30. Mitotic regulation of the APC activator proteins CDC20 and CDH1.
    Mol Biol Cell. 2000 May;11(5):1555-69 PMID: 10793135
  31. Phosphorylation of Cdc20/fizzy negatively regulates the mammalian cyclosome/APC in the mitotic checkpoint.
    Biochem Biophys Res Commun. 2000 May 10;271(2):299-304 PMID: 10799291
  32. Cdc14 activates cdc15 to promote mitotic exit in budding yeast.
    Curr Biol. 2000 May 18;10(10):615-8 PMID: 10837230
  33. A mechanism for coupling exit from mitosis to partitioning of the nucleus.
    Cell. 2000 Jul 7;102(1):21-31 PMID: 10929710
  34. The budding yeast Cdc15 localizes to the spindle pole body in a cell-cycle-dependent manner.
    Mol Cell Biol Res Commun. 1999 Sep-Dec;2(3):178-84 PMID: 10662594
  35. Anaphase spindle position is monitored by the BUB2 checkpoint.
    Nat Cell Biol. 2000 Aug;2(8):556-8 PMID: 10934478
  36. The Bub2p spindle checkpoint links nuclear migration with mitotic exit.
    Mol Cell. 2000 Jul;6(1):1-10 PMID: 10949022
  37. The budding yeast Dbf2 protein kinase localises to the centrosome and moves to the bud neck in late mitosis.
    J Cell Sci. 2000 Oct;113 Pt 19:3399-408 PMID: 10984431
  38. The spindle checkpoint of Saccharomyces cerevisiae responds to separable microtubule-dependent events.
    Curr Biol. 2000 Nov 2;10(21):1375-8 PMID: 11084338
  39. The Bfa1/Bub2 GAP complex comprises a universal checkpoint required to prevent mitotic exit.
    Curr Biol. 2000 Nov 2;10(21):1379-82 PMID: 11084339
  40. Nud1p links astral microtubule organization and the control of exit from mitosis.
    EMBO J. 2000 Dec 1;19(23):6475-88 PMID: 11101520
  41. Asymmetric spindle pole localization of yeast Cdc15 kinase links mitotic exit and cytokinesis.
    Curr Biol. 2001 Mar 6;11(5):345-50 PMID: 11267871
  42. A novel functional domain of Cdc15 kinase is required for its interaction with Tem1 GTPase in Saccharomyces cerevisiae.
    Genetics. 2001 Apr;157(4):1437-50 PMID: 11290702
  43. Characterization of the Net1 cell cycle-dependent regulator of the Cdc14 phosphatase from budding yeast.
    J Biol Chem. 2001 Jun 15;276(24):21924-31 PMID: 11274204
  44. Destruction of the CDC28/CLB mitotic kinase is not required for the metaphase to anaphase transition in budding yeast.
    EMBO J. 1993 May;12(5):1969-78 PMID: 8491189
  45. A multicopy suppressor gene of the Saccharomyces cerevisiae G1 cell cycle mutant gene dbf4 encodes a protein kinase and is identified as CDC5.
    Mol Cell Biol. 1993 Jul;13(7):4445-57 PMID: 8321244
  46. The Dbf2 and Dbf20 protein kinases of budding yeast are activated after the metaphase to anaphase cell cycle transition.
    EMBO J. 1994 Mar 1;13(5):1103-13 PMID: 8131744
  47. The yeast TEM1 gene, which encodes a GTP-binding protein, is involved in termination of M phase.
    Mol Cell Biol. 1994 Nov;14(11):7476-82 PMID: 7935462
  48. Isolation of a CDC25 family gene, MSI2/LTE1, as a multicopy suppressor of ira1.
    Yeast. 1994 Apr;10(4):451-61 PMID: 7941731
  49. P40SDB25, a putative CDK inhibitor, has a role in the M/G1 transition in Saccharomyces cerevisiae.
    Genes Dev. 1994 Jul 15;8(14):1640-53 PMID: 7958845
  50. Use of polymerase chain reaction epitope tagging for protein tagging in Saccharomyces cerevisiae.
    Yeast. 1995 Oct;11(13):1265-74 PMID: 8553697
  51. Movement of cortical actin patches in yeast.
    J Cell Biol. 1996 Mar;132(5):861-70 PMID: 8603918
  52. Anaphase initiation in Saccharomyces cerevisiae is controlled by the APC-dependent degradation of the anaphase inhibitor Pds1p.
    Genes Dev. 1996 Dec 15;10(24):3081-93 PMID: 8985178
  53. Yeast Hct1 is a regulator of Clb2 cyclin proteolysis.
    Cell. 1997 Aug 22;90(4):683-93 PMID: 9288748
Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1059-1524
DOI
10.1091/mbc.12.10.2961
Published
2001-10-00
Pages
2961-74
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC60148
Subset
IM
Grants
NIGMS NIH HHS · R01 GM056800 · United States
NIGMS NIH HHS · R29 GM056800 · United States
NIGMS NIH HHS · GM-56800 · United States
External Links
PubMed source DOI: 10.1091/mbc.12.10.2961 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2