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Potential roles for autophosphorylation, kinase activity, and abundance of a CDK-activating kinase (Ee;CDKF;1) during growth in leafy spurge

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Abstract

Leafy spurge (Euphorbia esula L.) is a deep-rooted perennial weed that propagates both by seeds and underground adventitious buds located on the crown and roots. To enhance our understanding of growth and development during seed germination and vegetative propagation, a leafy spurge gene (Accession No. AF230740) encoding a CDK-activating kinase (Ee;CDKF;1) involved in cell-cycle progression was identified, and its function was confirmed based on its ability to rescue a yeast temperature-sensitive CAK mutant (GF2351) and through in vitro kinase assays. Site-directed mutagenesis of Ee;CDKF;1 indicated that two threonine residues (Thr291 and Thr296) were mutually responsible for intra-molecular autophosphorylation and for phosphorylating its substrate protein, cyclin-dependent kinase (CDK). Polyclonal antibodies generated against the Ee;CDKF;1 protein or against a phosphorylated Ee;CDKF;1 peptide [NERYGSL(pT)SC] were used to examine abundance and phosphorylation of CDKF;1 during seed germination and bud growth. The levels of CDKF;1 were lower in dry or imbibed seeds than in germinating seeds or seedlings. Differences in CDKF;1 were also observed during adventitious bud development; small buds appeared to have greater levels of CDKF;1 than large buds. Similar patterns of CDKF;1 expression were detected with either the polyclonal antibody developed using the CDKF;1 protein or the phosphorylated peptide. These results indicated that Thr291 is constitutively phosphorylated in vivo and associated with Ee;CDKF;1 activity. Our results further suggest that a certain level of CDKF;1 activity is maintained in most tissues and may be an important phenomenon for enzymes that regulate early steps in cell-cycle signaling pathways.

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Abbreviations

CDK:

Cyclin-dependent kinase

CAK:

CDK-activating kinase

CTD:

C-terminal domain of the largest subunit of RNA polymerase II

MBP:

Maltose-binding protein

GST:

Glutathione S-transferase

References

  • Anderson JV, Horvath DP (2001) Random sequencing of cDNAs and identification of mRNAs. Weed Sci 49:590–597

    Article  CAS  Google Scholar 

  • Bewley JD (1997) Seed germination and dormancy. Plant Cell 9:1055–1066

    Article  PubMed  CAS  Google Scholar 

  • den Boer BGW, Murray JAH (2000) Control of plant growth and development through manipulation of cell-cycle genes. Curr Opin Biotech 11:138–145

    Article  Google Scholar 

  • Buck V, Russell P, Millar JBA (1995) Identification of a cdk-activating kinase in fission yeast. EMBO J 14:6173–6183

    PubMed  CAS  Google Scholar 

  • Campbell MA, Suttle JC, Sell TW (1996) Changes in cell cycle status and expression of p34cdc2 kinase during potato tuber meristem dormancy. Physiol Plant 98:743–752

    Article  CAS  Google Scholar 

  • Chao WS, Serpe MD, Anderson JV, Gesch RW, Horvath DP (2006) Sugars, hormones, and environment affect the dormancy status in underground adventitious buds of leafy spurge (Euphorbia esula). Weed Sci 54:59–68

    Article  CAS  Google Scholar 

  • Cismowski MJ, Laff GM, Solomon MJ, Reed SI (1995) 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 15:2983–2992

    PubMed  CAS  Google Scholar 

  • Damagnez V, Mäkelä TP, Cottarel G (1995) Schizosaccharomyces pombe Mop1-Mcs2 is related to mammalian CAK. EMBO J 14:6164–6172

    PubMed  CAS  Google Scholar 

  • Darbon JM, Devault A, Taviaux S, Fesquet D, Martinez AM, Galas S, Cavadore JC, Doree M, Blanchard JM (1994) Cloning, expression and subcellular localization of the human homolog of p40MO15 catalytic subunit of cdk-activating kinase. Oncogene 9:3127–3138

    PubMed  CAS  Google Scholar 

  • De Castro RD, Hilhorst HWM (2000) Dormancy, germination and the cell cycle in developing and imbibing tomato seeds. Rev Bras Fisiol Veg 12:105–136

    Google Scholar 

  • De Veylder L, Beeckman T, Beemster GTS, Krols L, Terras F, Landrieu I, Van Der Schueren E, Maes S, Naudts M, Inz D (2001) Functional analysis of cyclin-dependent kinase inhibitors of Arabidopsis. Plant Cell 13:1653–1667

    Article  PubMed  Google Scholar 

  • Devault A, Martinez AM, Fesquet D, Labbe JC, Morin N, Tassan JP, Nigg EA, Cavadore JC, Doree M (1995) MAT1 (‘menage a trois’): a new RING finger protein subunit stabilizing cyclin H-cdk7 complexes in starfish and Xenopus CAK. EMBO J 14:5027–5036

    PubMed  CAS  Google Scholar 

  • Devitt ML, Stafstrom JP (1995) Cell cycle regulation during growth-dormancy cycles in pea axillary buds. Plant Mol Biol 29:255–265

    Article  PubMed  CAS  Google Scholar 

  • Dewitte W, Murray JAH (2003) The plant cell cycle. Annu Rev Plant Biol 54:235–264

    Article  PubMed  CAS  Google Scholar 

  • Enke DA, Kaldis P, Holmes JK, Solomon MJ (1999) The CDK-activating kinase (Cak1p) from budding yeast has an unusual ATP-binding pocket. J Biol Chem 274:1949–1956

    Article  PubMed  CAS  Google Scholar 

  • Espinoza FH, Farrell A, Erdjument-Bromage H, Tempst P, Morgan DO (1996) A cyclin-dependent kinase-activating kinase (CAK) in budding yeast unrelated to vertebrate CAK. Science 273:1714–1717

    Article  PubMed  CAS  Google Scholar 

  • Fabian-Marwedel T, Umeda M, Sauter M (2002) The rice cyclin-dependent kinase-activating kinase R2 regulates S-phase progression. Plant Cell 14:197–210

    Article  PubMed  CAS  Google Scholar 

  • Faye G, Simon M, Valay JG, Fesquet D, Facca C (1997) Rig2, a RING finger protein that interacts with the Kin28/Ccl1 CTD kinase in yeast. Mol Gen Genet 255:460–466

    Article  PubMed  CAS  Google Scholar 

  • Feaver WJ, Svejstrup JQ, Henry NL, Kornberg RD (1994) Relationship of CDK-activating kinase and RNA polymerase II CTD kinase TFIIH/TFIIK. Cell 79:1103–1109

    Article  PubMed  CAS  Google Scholar 

  • Feaver WJ, Henry NL, Wang Z, Wu X, Svejstrup JQ, Bushnell DA (1997) Genes for Tfb2, Tfb3, and Tfb4 subunits of yeast transcription/repair factor IIH. J Biol Chem 272:19319–19327

    Article  PubMed  CAS  Google Scholar 

  • Fesquet D, Labbe JC, Derancourt J, Capony JP, Galas S, Girard F, Lorca T, Shuttleworth J, Dorée M, Cavadore JC (1993) 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 12:3111–3121

    PubMed  CAS  Google Scholar 

  • Fisher RP, Jin P, Chamberlin HM, Morgan DO (1995) Alternative mechanisms of CAK assembly require an assembly factor or an activating kinase. Cell 83:47–57

    Article  PubMed  CAS  Google Scholar 

  • Fisher RP, Morgan DO (1994) A novel cyclin associates with MO15/CDK7 to form the CDK-activating kinase. Cell 78:713–724

    Article  PubMed  CAS  Google Scholar 

  • Freeley M, Volkov Y, Kelleher D, Long A (2005) Stimulus-induced phosphorylation of PKC θ at the C-terminal hydrophobic-motif in human T lymphocytes. Biochem Biophy Res Commun 334:619–630

    Article  CAS  Google Scholar 

  • Garret S, Barton WA, Knights R, Jin P, Morgan DO, Fisher RP (2001) Reciprocal activation by cyclin-dependent kinases 2 and 7 is directed by substrate specificity determinants outside the T loop. Mol Cell Biol 22:88–99

    Article  Google Scholar 

  • Gietz D, St Jean A, Woods RA, Schiestl RH (1992) Improved method for high efficiency transformation of intact yeast cells. Nucleic Acids Res 20:1425

    Article  PubMed  CAS  Google Scholar 

  • Gutierrez C, Ramirez-Parra E, Castellano MM, del Pozo JC (2002) G(1) to S transition: more than a cell cycle engine switch. Curr Opin Plant Biol 5:480–486

    Article  PubMed  CAS  Google Scholar 

  • Hagopian JC, Kirtley MP, Stevenson LM, Gergis RM, Russo AA, Pavletich NP, Parsons SM, Lew J (2001) Kinetic basis for activation of CDK2/cyclin A by phosphorylation. J Biol Chem 276:275–280

    Article  PubMed  CAS  Google Scholar 

  • Hata S (1991) cDNA cloning of a novel cdc2 +/CDC28-related protein kinase from rice. FEBS Lett 279:149–152

    Article  PubMed  CAS  Google Scholar 

  • Hermand D, Pihlak A, Westerling T, Damagnez V, Vandenhaute J, Cottarel G, Mäkelä TP (1998) Fission yeast Csk1 is a CAK-activating kinase (CAKAK). EMBO J 17:7230–7238

    Article  PubMed  CAS  Google Scholar 

  • Hermand D, Westerling T, Pihlak A, Thuret JY, Vallenius T, Tiainen M, Vandenhaute J, Cottarel G, Mann C, Mäkelä TP (2001) Specificity of Cdk activation in vivo by the two Caks Mcs6 and Csk1 in fission yeast. EMBO J 20:82–90

    Article  PubMed  CAS  Google Scholar 

  • Horvath DP, Anderson JV, Chao WS, Foley ME (2003). Knowing when to grow: signals regulating bud dormancy. Trends Plant Sci 8:534–540

    Article  PubMed  CAS  Google Scholar 

  • Huntley RP, Murray AH (1999) The plant cell cycle. Curr Opin Plant Biol 2:440–446

    Article  PubMed  CAS  Google Scholar 

  • Jeffery PD, Russo AA, Polyak K, Gibbs E, Hurwitz J, Massagué J, Pavletich NP (1995) Mechanism of cdk activation revealed by the structure of a cyclin A-CDK2 complex. Nature 376:313–320

    Article  Google Scholar 

  • Kaldis P, Sutton A, Solomon MJ (1996) The Cdk-activating kinase (CAK) from budding yeast. Cell 86:553–564

    Article  PubMed  CAS  Google Scholar 

  • Kaldis P, Pitluk ZW, Bany IA, Enke DA, Wagner M, Winter E, Solomon MJ (1998a) Localization and regulation of the cdk-activating kinase (Cak1p) from budding yeast. J Cell Sci 111:3585–3596

    CAS  Google Scholar 

  • Kaldis P, Russo AA, Chou HS, Pavletich NP, Solomon MJ (1998b) Human and yeast cdk-activating kinases (CAKs) display distinct substrate specificities. Mol Biol Cell 9:2545–2560

    CAS  Google Scholar 

  • Kaldis P (1999) The cdk-activating kinase (CAK): from yeast to mammals. Cell Mol Life Sci 55:284–296

    Article  PubMed  CAS  Google Scholar 

  • Kucera B, Cohn MA, Leubner-Metzger G (2005) Plant hormone interactions during seed dormancy release and germination. Seed Sci Res 15:281–307

    Article  CAS  Google Scholar 

  • Labbé JC, Martinez AM, Fesquet D, Capony JP, Darbon JM, Derancourt J, Devault A, Morin N, Cavadore JC, Dorée M (1994) p40MO15 associates with a p36 subunit and requires both nuclear translocation and Thr176 phosphorylation to generate cdk-activating kinase activity in Xenopus oocytes. EMBO J 13:5155–5164

    PubMed  Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Article  PubMed  CAS  Google Scholar 

  • Larochelle S, Pandur J, Fisher RP, Salz HK, Suter B (1998) Cdk7 is essential for mitosis and for in vivo Cdk-activating kinase activity. Genes Dev 12:370–381

    PubMed  CAS  Google Scholar 

  • Lee KM, Saiz JE, Barton WA, Fisher RP (1999) Cdc2 activation in fission yeast depends on Mcs6 and Csk1, two partially redundant Cdk-activating kinases (CAKs). Curr Biol 9:441–444

    Article  PubMed  CAS  Google Scholar 

  • Martinez AM, Afshar M, Martin F, Cavadore JC, Labbé JC, Dorée M (1997) Dual phosphorylation of the T-loop in cdk: its role in controlling cyclin H binding and CAK activity. EMBO J 16:343–354

    Article  PubMed  CAS  Google Scholar 

  • Morgan DO (1995) Principles of CDK regulation. Nature 374:131–134

    Article  PubMed  CAS  Google Scholar 

  • Morgan DO (1997) Cyclin-dependent kinases: engines, clocks, and microprocessors. Annu Rev Cell Dev Biol 13:261–291

    Article  PubMed  CAS  Google Scholar 

  • Mäkelä TP, Tassan JP, Nigg EA, Frutiger S, Hughes GJ, Weinberg RA (1994) A cyclin associated with the CDK-activating kinase MO15. Nature 371:254–257

    Article  PubMed  Google Scholar 

  • Poon RYC, Yamashita K, Adamczewski JP, Hunt T, Shuttleworth J (1993) The cdc2-related protein p40MO15 is the catalytic subunit of a protein kinase that can activate p33cdk2 and p34cdc2. EMBO J 12:3123–3132

    PubMed  CAS  Google Scholar 

  • Raju MVS, Steeves TA, Coupland RT (1964) On the regeneration of root fragments of leafy spurge (Euphorbia esula L.). Weed Res 4:2–11

    Article  Google Scholar 

  • Roy R, Adamczeski JP, Seroz T, Vermeulen W, Tassan JP, Schaeffer L, Nigg EA, Hoeijmakers JHJ, Egly JM (1994) The MO15 cell cycle kinase is associated with the TFIIH transcription–DNA repair factor. Cell 79:1093–1101

    Article  PubMed  CAS  Google Scholar 

  • Russo AA, Jeffrey PD, Pavletich NP (1996) Structural basis of cyclin-dependent kinase activation by phosphorylation. Nature Struct Biol 3:696–700

    Article  PubMed  CAS  Google Scholar 

  • Serizawa H, Makela TP, Conaway JW, Conaway RC, Weinberg RA, Young RA (1995) Association of Cdk-activating kinase subunits with transcription factor TFIIH. Nature 374:280–282

    Article  PubMed  CAS  Google Scholar 

  • Shiekhattar R, Mermelstein F, Fisher RP, Drapkin R, Dynlacht B, Wessling HC, Morgan DO, Reinberg D (1995) Cdk-activating kinase complex is a component of human transcription factor TFIIH. Nature 374:283–287

    Article  PubMed  CAS  Google Scholar 

  • Shimizu-Sato S, Mori H (2001) Control of Outgrowth and Dormancy in Axillary Buds. Plant Physiol 127:1405–1413

    Article  PubMed  CAS  Google Scholar 

  • Shimotohno A, Matsubayashi S, Yamaguchi M, Uchimiya H, Umeda M (2003) Differential phosphorylation activities of CDK-activating kinases in Arabidopsis thaliana. FEBS Lett 534:69–74

    Article  PubMed  CAS  Google Scholar 

  • Shimotohno A, Umeda-Hara C, Bisova K, Uchimiya H, Umeda M (2004) The plant-specific kinase CDKF;1 is involved in activating phosphorylation of cyclin-dependent kinase-activating kinases in Arabidopsis. Plant Cell 16:2954–2966

    Article  PubMed  CAS  Google Scholar 

  • Shimotohno A, Ohno R, Bisova K, Sakaguchi N, Huang J, Koncz C, Uchimiya H, Umeda M (2006) Diverse phosphoregulatory mechanisms controlling cyclin-dependent kinase-activating kinases in Arabidopsis. Plant J 47:701–710

    Article  PubMed  CAS  Google Scholar 

  • Solomon MJ, Harper JW, Shuttleworth J (1993) CAK, the p34cdc2 activating kinase contains a protein identical or closely related to p40MO15. EMBO J 12:3133–3142

    PubMed  CAS  Google Scholar 

  • Tassan JP, Jacquenoud M, Fry AM, Frutiger S, Hughes GJ, Nigg EA (1995) In vitro assembly of a functional human CDK7-cyclin H complex requires MAT1, a novel 36 kDa RING finger protein. EMBO J 14:5608–5617

    PubMed  CAS  Google Scholar 

  • Thuret JY, Valay JG, Faye G, Mann C (1996) Civ1 (CAK in vivo), a novel Cdk-activating kinase. Cell 86:565–576

    Article  PubMed  CAS  Google Scholar 

  • Tsakraklides V, Solomon MJ (2002) Comparison of Cak1p-like cyclin-dependent kinase-activating kinases. J Biol Chem 277:33482–33489

    Article  PubMed  CAS  Google Scholar 

  • Umeda M, Bhalerao RP, Schell J, Uchimiya H, Koncz C (1998) A distinct cylcin-dependent kinase-activating kinase of Arabidopsis thaliana. Proc Natl Acad Sci USA 95:5021–5026

    Article  PubMed  CAS  Google Scholar 

  • Valay JG, Simon M, Faye G (1993) The Kin28 protein kinase is associated with a cyclin in Saccharomyces cerevisiae. J Mol Biol 234:307–310

    Article  PubMed  CAS  Google Scholar 

  • Valay JG, Simon M, Dubois MF, Bensaude O, Facca C, Faye G (1995) The KIN28 gene is required both for RNA polymerase II mediated transcription and phosphorylation of the Rpb1p CTD. J Mol Biol 249:535–544

    Article  PubMed  CAS  Google Scholar 

  • Wang CS, Walling LL, Eckard KJ, Lord EM (1992) Patterns of protein accumulation in developing anthers of Lilium Longiflorum correlate with histological events. Am J Bot 79:118–127

    Article  CAS  Google Scholar 

  • Yamaguchi M, Umeda M, Uchimiya H (1998) A rice homolog of Cdk7/MO15 phosphorylates both cyclin-dependent protein kinases and the carboxy-terminal domain of RNA polymerase II. Plant J 16:613–619

    Article  PubMed  CAS  Google Scholar 

  • Yee A, Nichols MA, Wu L, Hall FL, Kobayashi R, Xiong Y (1995) Molecular cloning of CDK7-associated human MAT1, a cyclin-dependent kinase-activating kinase (CAK) assembly factor. Cancer Res 55:6058–6062

    PubMed  CAS  Google Scholar 

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Acknowledgments

The authors acknowledge Wayne Sargent, USDA-ARS, Fargo, ND, for his technical assistance. We thank Dr. Carl Mann for providing us the yeast GF2351.

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Authors and Affiliations

  1. USDA-Agricultural Research Service, Biosciences Research Laboratory, 1605 Albrecht Blvd., Fargo, ND, 58105-5674, USA

    Wun S. Chao, Weilin L. Shelver & James V. Anderson

  2. Department of Biology, Boise State University, Boise, ID, 83725, USA

    Marcelo D. Serpe

  3. Department of Plant Sciences, North Dakota State University, Fargo, ND, 58105, USA

    Ying Jia

  4. Graduate School of Biological Sciences, Nara Institute of Science and Technology, Ikoma, Nara, 630-0101, Japan

    Masaaki Umeda

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  1. Wun S. Chao
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  2. Marcelo D. Serpe
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  3. Ying Jia
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  4. Weilin L. Shelver
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  5. James V. Anderson
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  6. Masaaki Umeda
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Correspondence to Wun S. Chao.

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Chao, W.S., Serpe, M.D., Jia, Y. et al. Potential roles for autophosphorylation, kinase activity, and abundance of a CDK-activating kinase (Ee;CDKF;1) during growth in leafy spurge. Plant Mol Biol 63, 365–379 (2007). https://doi.org/10.1007/s11103-006-9094-y

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