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PMID: 11226158 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Specificity of Cdk activation in vivo by the two Caks Mcs6 and Csk1 in fission yeast.

The EMBO journal ·Vol. 20 ·No. 1-2 ·2001-01-15 ·Pages 82-90

Hermand D, Westerling T, Pihlak A, Thuret JY, Vallenius T, Tiainen M, Vandenhaute J, Cottarel G, Mann C, Mäkelä TP

Abstract

Activating phosphorylation of cyclin-dependent kinases (Cdks) is mediated by at least two structurally distinct types of Cdk-activating kinases (Caks): the trimeric Cdk7-cyclin H-Mat1 complex in metazoans and the single-subunit Cak1 in budding yeast. Fission yeast has both Cak types: Mcs6 is a Cdk7 ortholog and Csk1 a single-subunit kinase. Both phosphorylate Cdks in vitro and rescue a thermosensitive budding yeast CAK1 strain. However, this apparent redundancy is not observed in fission yeast in vivo. We have identified mutants that exhibit phenotypes attributable to defects in either Mcs6-activating phosphorylation or in Cdc2-activating phosphorylation. Mcs6, human Cdk7 and budding yeast Cak1 were all active as Caks for Cdc2 when expressed in fission yeast. Although Csk1 could activate Mcs6, it was unable to activate Cdc2. Biochemical experiments supported these genetic results: budding yeast Cak1 could bind and phosphorylate Cdc2 from fission yeast lysates, whereas fission yeast Csk1 could not. These results indicate that Mcs6 is the direct activator of Cdc2, and Csk1 only activates Mcs6. This demonstrates in vivo specificity in Cdk activation by Caks.

MeSH Terms
Cloning, Molecular Cyclin-Dependent Kinases/metabolism Enzyme Activation Genetic Complementation Test Glutathione Transferase/genetics Humans Open Reading Frames Phenotype Phosphorylation Protein Kinases/genetics,metabolism Protein Serine-Threonine Kinases/genetics,metabolism Recombinant Fusion Proteins/metabolism Schizosaccharomyces/enzymology,genetics,growth & development Schizosaccharomyces pombe Proteins Substrate Specificity
Chemicals
Recombinant Fusion Proteins Schizosaccharomyces pombe Proteins Glutathione Transferase Protein Kinases Sck1 protein, S pombe Protein Serine-Threonine Kinases Cyclin-Dependent Kinases cyclin-dependent kinase-activating kinase
Authors & Affiliations
10 authors, click to expand affiliations / ORCID
Hermand D
Haartman Institute & Biocentrum Helsinki, University of Helsinki, 00014 Helsinki, HUCH Laboratory Diagnostics, 00029 HYKS, Finland.
Westerling T
Pihlak A
Thuret J Y
Vallenius T
Tiainen M
Vandenhaute J
Cottarel G
Mann C
Mäkelä T P
References (64)
64 references, click to expand
  1. Human cyclin-dependent kinase-activating kinase exists in three distinct complexes.
    Proc Natl Acad Sci U S A. 1996 Jun 25;93(13):6488-93 PMID: 8692842
  2. Activating phosphorylation of the Kin28p subunit of yeast TFIIH by Cak1p.
    Mol Cell Biol. 1999 Jul;19(7):4774-87 PMID: 10373527
  3. The Cdk-activating kinase (CAK) from budding yeast.
    Cell. 1996 Aug 23;86(4):553-64 PMID: 8752210
  4. Civ1 (CAK in vivo), a novel Cdk-activating kinase.
    Cell. 1996 Aug 23;86(4):565-76 PMID: 8752211
  5. A cyclin-dependent kinase-activating kinase (CAK) in budding yeast unrelated to vertebrate CAK.
    Science. 1996 Sep 20;273(5282):1714-7 PMID: 8781234
  6. Alternative mechanisms of CAK assembly require an assembly factor or an activating kinase.
    Cell. 1995 Oct 6;83(1):47-57 PMID: 7553872
  7. MAT1 ('menage à trois') a new RING finger protein subunit stabilizing cyclin H-cdk7 complexes in starfish and Xenopus CAK.
    EMBO J. 1995 Oct 16;14(20):5027-36 PMID: 7588631
  8. In vitro assembly of a functional human CDK7-cyclin H complex requires MAT1, a novel 36 kDa RING finger protein.
    EMBO J. 1995 Nov 15;14(22):5608-17 PMID: 8521818
  9. Schizosaccharomyces pombe Mop1-Mcs2 is related to mammalian CAK.
    EMBO J. 1995 Dec 15;14(24):6164-72 PMID: 8557036
  10. Identification of a cdk-activating kinase in fission yeast.
    EMBO J. 1995 Dec 15;14(24):6173-83 PMID: 8557037
  11. Isolation and characterization of two human transcription factor IIH (TFIIH)-related complexes: ERCC2/CAK and TFIIH.
    Proc Natl Acad Sci U S A. 1996 Jun 25;93(13):6482-7 PMID: 8692841
  12. A quantitative model for the cdc2 control of S phase and mitosis in fission yeast.
    Trends Genet. 1996 Sep;12(9):345-50 PMID: 8855663
  13. Genes for Tfb2, Tfb3, and Tfb4 subunits of yeast transcription/repair factor IIH. Homology to human cyclin-dependent kinase activating kinase and IIH subunits.
    J Biol Chem. 1997 Aug 1;272(31):19319-27 PMID: 9235928
  14. The Cak1p protein kinase is required at G1/S and G2/M in the budding yeast cell cycle.
    Genetics. 1997 Sep;147(1):57-71 PMID: 9286668
  15. Rig2, a RING finger protein that interacts with the Kin28/Ccl1 CTD kinase in yeast.
    Mol Gen Genet. 1997 Aug;255(5):460-6 PMID: 9294030
  16. Is Cdk7/cyclin H/MAT1 the genuine cdk activating kinase in cycling Xenopus egg extracts?
    Oncogene. 1997 Sep;15(11):1303-7 PMID: 9315098
  17. Cyclin-dependent kinases: engines, clocks, and microprocessors.
    Annu Rev Cell Dev Biol. 1997;13:261-91 PMID: 9442875
  18. Cdk7 is essential for mitosis and for in vivo Cdk-activating kinase activity.
    Genes Dev. 1998 Feb 1;12(3):370-81 PMID: 9450931
  19. Cyclin B proteolysis and the cyclin-dependent kinase inhibitor rum1p are required for pheromone-induced G1 arrest in fission yeast.
    Mol Biol Cell. 1998 Jun;9(6):1309-21 PMID: 9614176
  20. Transcriptional activation independent of TFIIH kinase and the RNA polymerase II mediator in vivo.
    Nature. 1998 May 28;393(6683):389-92 PMID: 9620805
  21. Human cyclin K, a novel RNA polymerase II-associated cyclin possessing both carboxy-terminal domain kinase and Cdk-activating kinase activity.
    Mol Cell Biol. 1998 Jul;18(7):4291-300 PMID: 9632813
  22. Regulation of CDKs by phosphorylation.
    Results Probl Cell Differ. 1998;22:79-109 PMID: 9670320
  23. Temporal regulation of RNA polymerase II by Srb10 and Kin28 cyclin-dependent kinases.
    Mol Cell. 1998 Jul;2(1):43-53 PMID: 9702190
  24. Dominant-negative mutants reveal a role for the Cdk7 kinase at the mid-blastula transition in Drosophila embryos.
    EMBO J. 2000 Apr 3;19(7):1567-75 PMID: 10747025
  25. Analysis of CAK activities from human cells.
    Eur J Biochem. 2000 Jul;267(13):4213-21 PMID: 10866826
  26. The cell cycle control gene cdc2+ of fission yeast encodes a protein kinase potentially regulated by phosphorylation.
    Cell. 1986 Apr 25;45(2):261-8 PMID: 3516412
  27. Vectors for the construction of gene banks and the integration of cloned genes in Schizosaccharomyces pombe and Saccharomyces cerevisiae.
    Plasmid. 1986 Mar;15(2):156-8 PMID: 3010354
  28. KIN28, a yeast split gene coding for a putative protein kinase homologous to CDC28.
    EMBO J. 1986 Oct;5(10):2697-701 PMID: 3536482
  29. Interaction between cdc13+ and cdc2+ in the control of mitosis in fission yeast; dissociation of the G1 and G2 roles of the cdc2+ protein kinase.
    EMBO J. 1987 Nov;6(11):3441-7 PMID: 3428262
  30. Regulated expression and phosphorylation of a possible mammalian cell-cycle control protein.
    Nature. 1988 Jun 16;333(6174):676-9 PMID: 3287181
  31. Invariant phosphorylation of the Saccharomyces cerevisiae Cdc28 protein kinase.
    Mol Cell Biol. 1988 Jul;8(7):2976-9 PMID: 3043202
  32. Control of the yeast cell cycle is associated with assembly/disassembly of the Cdc28 protein kinase complex.
    Cell. 1988 Sep 23;54(7):1061-72 PMID: 3046752
  33. A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae.
    Genetics. 1989 May;122(1):19-27 PMID: 2659436
  34. cdc2 and the regulation of mitosis: six interacting mcs genes.
    Genetics. 1989 Aug;122(4):773-82 PMID: 2474475
  35. p40MO15, a cdc2-related protein kinase involved in negative regulation of meiotic maturation of Xenopus oocytes.
    EMBO J. 1990 Oct;9(10):3233-40 PMID: 2209545
  36. Molecular genetic analysis of fission yeast Schizosaccharomyces pombe.
    Methods Enzymol. 1991;194:795-823 PMID: 2005825
  37. Biological activity of the mammalian RAP genes in yeast.
    Cell Regul. 1990 Sep;1(10):763-9 PMID: 1983087
  38. Phosphorylation at Thr167 is required for Schizosaccharomyces pombe p34cdc2 function.
    EMBO J. 1991 Nov;10(11):3297-309 PMID: 1655416
  39. Role of phosphorylation in p34cdc2 activation: identification of an activating kinase.
    Mol Biol Cell. 1992 Jan;3(1):13-27 PMID: 1532335
  40. Thiamine-repressible expression vectors pREP and pRIP for fission yeast.
    Gene. 1993 Jan 15;123(1):127-30 PMID: 8422996
  41. Characterization of the fission yeast mcs2 cyclin and its associated protein kinase activity.
    EMBO J. 1993 Apr;12(4):1723-32 PMID: 8467814
  42. 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):3111-21 PMID: 8344251
  43. 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):3123-32 PMID: 8393783
  44. CAK, the p34cdc2 activating kinase, contains a protein identical or closely related to p40MO15.
    EMBO J. 1993 Aug;12(8):3133-42 PMID: 8344252
  45. The kin28 protein kinase is associated with a cyclin in Saccharomyces cerevisiae.
    J Mol Biol. 1993 Nov 20;234(2):307-10 PMID: 8230216
  46. Molecular cloning and characterization of the Schizosaccharomyces pombe his3 gene for use as a selectable marker.
    Mol Gen Genet. 1994 Jan;242(2):169-76 PMID: 8159167
  47. A novel cyclin associates with MO15/CDK7 to form the CDK-activating kinase.
    Cell. 1994 Aug 26;78(4):713-24 PMID: 8069918
  48. A cyclin associated with the CDK-activating kinase MO15.
    Nature. 1994 Sep 15;371(6494):254-7 PMID: 8078587
  49. Cell cycle analysis of the activity, subcellular localization, and subunit composition of human CAK (CDK-activating kinase).
    J Cell Biol. 1994 Oct;127(2):467-78 PMID: 7929589
  50. The MO15 cell cycle kinase is associated with the TFIIH transcription-DNA repair factor.
    Cell. 1994 Dec 16;79(6):1093-101 PMID: 8001135
  51. Relationship of CDK-activating kinase and RNA polymerase II CTD kinase TFIIH/TFIIK.
    Cell. 1994 Dec 16;79(6):1103-9 PMID: 8001136
  52. Regulatable promoters of Saccharomyces cerevisiae: comparison of transcriptional activity and their use for heterologous expression.
    Nucleic Acids Res. 1994 Dec 25;22(25):5767-8 PMID: 7838736
  53. Association of Cdk-activating kinase subunits with transcription factor TFIIH.
    Nature. 1995 Mar 16;374(6519):280-2 PMID: 7885450
  54. Cdk-activating kinase complex is a component of human transcription factor TFIIH.
    Nature. 1995 Mar 16;374(6519):283-7 PMID: 7533895
  55. KIN28 encodes a C-terminal domain kinase that controls mRNA transcription in Saccharomyces cerevisiae but lacks cyclin-dependent kinase-activating kinase (CAK) activity.
    Mol Cell Biol. 1995 Jun;15(6):2983-92 PMID: 7760796
  56. A kinase-deficient transcription factor TFIIH is functional in basal and activated transcription.
    Proc Natl Acad Sci U S A. 1995 May 23;92(11):5174-8 PMID: 7761469
  57. The KIN28 gene is required both for RNA polymerase II mediated transcription and phosphorylation of the Rpb1p CTD.
    J Mol Biol. 1995 Jun 9;249(3):535-44 PMID: 7783209
  58. Heterologous modules for efficient and versatile PCR-based gene targeting in Schizosaccharomyces pombe.
    Yeast. 1998 Jul;14(10):943-51 PMID: 9717240
  59. Human and yeast cdk-activating kinases (CAKs) display distinct substrate specificities.
    Mol Biol Cell. 1998 Sep;9(9):2545-60 PMID: 9725911
  60. Cak1 is required for Kin28 phosphorylation and activation in vivo.
    Mol Cell Biol. 1998 Nov;18(11):6365-73 PMID: 9774652
  61. Fission yeast Csk1 is a CAK-activating kinase (CAKAK).
    EMBO J. 1998 Dec 15;17(24):7230-8 PMID: 9857180
  62. The cdk-activating kinase (CAK): from yeast to mammals.
    Cell Mol Life Sci. 1999 Feb;55(2):284-96 PMID: 10188587
  63. Cdc2 activation in fission yeast depends on Mcs6 and Csk1, two partially redundant Cdk-activating kinases (CAKs).
    Curr Biol. 1999 Apr 22;9(8):441-4 PMID: 10226032
  64. Dephosphorylation of threonine 169 of Cdc28 is not required for exit from mitosis but may be necessary for start in Saccharomyces cerevisiae.
    Mol Cell Biol. 1996 Aug;16(8):4573-83 PMID: 8754858
Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/20.1.82
Published
2001-01-15
Pages
82-90
Language
English
Region
England
NLM ID
8208664
PMCID
PMC140202
Subset
IM
External Links
PubMed source DOI: 10.1093/emboj/20.1.82 PMC Full Text View journal details
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