The Arabidopsis D-type cyclins CycD2 and CycD3 both interact in vivo with the PSTAIRE cyclin-dependent kinase Cdc2a but are differentially controlled.
D-type cyclins (CycD) play key roles in linking the Arabidopsis cell cycle to extracellular and developmental signals, but little is known of their regulation at the post-transcriptional level or of their cyclin-dependent kinase (CDK) partners. Using new antisera to CycD2 and CycD3, we demonstrate that the CDK partner of these Arabidopsis cyclins is the PSTAIRE-containing CDK Cdc2a. Previous analysis has shown that transcript levels of CycD2 and CycD3 are regulated in response to sucrose levels and that both their mRNA levels and kinase activity are induced with different kinetics during the G(1) phase of cells reentering the division cycle from quiescence. Here we analyze the protein levels and kinase activity of CycD2 and CycD3. We show that CycD3 protein and kinase activity parallel the abundance of its mRNA and that CycD3 protein is rapidly lost from cells in stationary phase or following sucrose removal. In contrast to both CycD3 and the regulation of its own mRNA levels, CycD2 protein is present at constant levels. CycD2 kinase activity is regulated by sequestration of CycD2 protein in a form inaccessible to immunoprecipitation and probably not complexed to Cdc2a.
Healy
J M
JM
Institute of Biotechnology, University of Cambridge, United Kingdom.
Menges
M
M
Doonan
J H
JH
Murray
J A
JA
D017360:Arabidopsis / Q000737:chemistry*; D029681:Arabidopsis Proteins; D015153:Blotting, Western; D016203:CDC2 Protein Kinase / Q000737:chemistry*; D056743:Cyclin D3; D018844:Cyclin-Dependent Kinases / Q000378:metabolism*; D016213:Cyclins / Q000378:metabolism*; D007700:Kinetics; D008954:Models, Biological; D010446:Peptide Fragments / Q000737:chemistry*; D011233:Precipitin Tests; D011485:Protein Binding; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000378:metabolism; D013395:Sucrose / Q000494:pharmacology; D013997:Time Factors
D029681:Arabidopsis Proteins; C486198:CycD2 protein, Arabidopsis; D056743:Cyclin D3; D016213:Cyclins; D010446:Peptide Fragments; D012333:RNA, Messenger; C062418:PSTAIR peptide; D013395:Sucrose; D016203:CDC2 Protein Kinase; C418966:CDC2A protein, Arabidopsis; D018844:Cyclin-Dependent Kinases