The cytokinin requirement for cell division in cultured Nicotiana plumbaginifolia cells can be satisfied by yeast Cdc25 protein tyrosine phosphatase: implications for mechanisms of cytokinin response and plant development.
Cultured cells of Nicotiana plumbaginifolia, when deprived of exogenous cytokinin, arrest in G2 phase prior to mitosis and then contain cyclin-dependent protein kinase (CDK) that is inactive because phosphorylated on tyrosine (Tyr). The action of cytokinin in stimulating the activation of CDK by removal of inhibitory phosphorylation from Tyr is not a secondary downstream consequence of other hormone actions but is the key primary effect of the hormone in its stimulation of cell proliferation, since cytokinin could be replaced by expression of cdc25, which encodes the main Cdc2 (CDK)-Tyr dephosphorylating enzyme of yeast (Saccharomyces cerevisiae). The cdc25 gene, under control of a steroid-inducible promoter, induced a rise in cdc25 mRNA, accumulation of p67(Cdc25) protein, and increase in Cdc25 phosphatase activity that was measured in vitro with Tyr-phosphorylated Cdc2 as substrate. Cdc25 phosphatase activity peaked during mitotic prophase at the time CDK activation was most rapid. Mitosis that was induced by cytokinin also involved increase in endogenous plant CDK Tyr phosphatase activity during prophase, therefore indicating that this is a normal part of plant mitosis. These results suggest a biochemical mechanism for several previously described transgene phenotypes in whole plants and suggest that a primary signal from cytokinin leading to progression through mitosis is the activation of CDK by dephosphorylation of Tyr.
Zhang
Kerong
K
Plant Cell Biology Group, Research School of Biological Sciences, Australian National University, Canberra, Australian Capital Territory 2601, Australia.
Diederich
Ludger
L
John
Peter C L
PC
D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism; D002478:Cells, Cultured; D003470:Culture Media / Q000737:chemistry; D018844:Cyclin-Dependent Kinases / Q000378:metabolism*; D003583:Cytokinins / Q000502:physiology*; D004789:Enzyme Activation; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D008938:Mitosis / Q000502:physiology*; D030821:Plants, Genetically Modified; D011499:Protein Processing, Post-Translational; D012441:Saccharomyces cerevisiae; D013997:Time Factors; D014026:Nicotiana / Q000166:cytology* / Q000235:genetics; D019076:Transgenes; D020666:ras-GRF1 / Q000235:genetics / Q000378:metabolism
D018797:Cell Cycle Proteins; D003470:Culture Media; D003583:Cytokinins; D005656:Fungal Proteins; D020666:ras-GRF1; D018844:Cyclin-Dependent Kinases