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Plant Physiol, July 2001, Vol. 126, pp. 1214-1223
Cell Cycle Regulation of Cyclin-Dependent Kinases in Tobacco
Cultivar Bright Yellow-2 Cells1
David A.
Sorrell,2
Margit
Menges,
J.M. Sandra
Healy,3
Yves
Deveaux,
Chinatsu
Amano,
Ya
Su,4
Hirofumi
Nakagami,
Atsuhiko
Shinmyo,
John H.
Doonan,
Masami
Sekine, and
James A.H.
Murray*
Institute of Biotechnology, University of Cambridge, Cambridge CB2
1QT, United Kingdom (D.A.S., M.M., J.M.S.H., Y.D., Y.S., M.S.,
J.A.H.M.); Graduate School of Biological Sciences, Nara Institute of
Science and Technology, Takayama 8916-5, Ikoma, Nara 630-010, Japan
(C.A., H.N., A.S., M.S.); and Department of Cell Biology, John Innes
Institute, Colney Lane, Norwich, NR4 7UH, United Kingdom
(J.H.D.)
Plants possess two major classes of cyclin-dependent kinases (CDK)
with cyclin-binding motifs PSTAIRE (CDK-a) and PPTA/TLRE (CDK-b).
Tobacco (Nicotiana tabacum L. cv Bright Yellow-2) cells are the most highly synchronizable plant culture, but no detailed analysis of CDK activities has been reported in this system. Here we
describe isolation of new PPTALRE CDKs (Nicta;CdkB1)
from Bright Yellow-2 cells and present detailed analysis of the mRNA,
protein and kinase activity levels of CdkB1, and the
PSTAIRE CDKA during the growth and cell cycles. CdkA and
CdkB1 transcripts are more abundant in exponential than
in stationary phase cells, but the two genes show strikingly different
regulation during the cell cycle. CdkA mRNA and protein
accumulate during G1 in cells re-entering the cell cycle, and
immunoprecipitated histone H1 kinase activity increases at the G1/S
boundary. Aphidicolin synchronized cells show the highest
CDKA-associated histone H1 kinase activity during S-G2 phases, although
CdkA mRNA and protein levels are not significantly regulated. In contrast, CdkB1 transcripts are present at
very low levels until S phase and CDKB1 protein and kinase activity is
almost undetectable in G1. CdkB1 mRNA accumulates
through S until M phase and its associated kinase activity peaks at the G2/M boundary, confirming that transcription of PPTALRE CDKs is cell
cycle regulated. We suggest that CDKA kinase activity likely plays
roles at the G1/S phase boundary, during S phase, and at the G2/M phase
transition, and that CDKB1 kinase activity is present only at
G2/M.
1
This work was supported in part by the
Biotechnology and Biological Sciences Research Council (to J.A.H.M. and
J.H.D.; studentships to D.A.S. and M.M.), by a Grant-in-Aid of
Scientific Research from the Ministry of Education, Science and
Culture, Japan (grant no. 12037213 to M.S.), and by Aventis CropScience.
2
Present address: Cardiff School of Biosciences, Cardiff
University, P.O. Box 915, Cardiff, CF10 3TL, UK.
3
Present address: Dipartimento di Genetica, IV Piano,
Torre A, Università di Milano, 20133 Milan, Italy.
4
Present address: Department of Molecular Genetics,
Wellcome Trust Centre for Molecular Mechanisms in Disease, University
of Cambridge, Addenbrooke's Hospital, Hills Road, Cambridge, CB2 2XY, UK.
*
Corresponding author; e-mail j.murray{at}biotech.cam.ac.uk; fax
44-1223-334162.
© 2001 American Society of Plant Physiologists
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