title: Suppression of WEE1 and stimulation of CDC25A correlates with endothelin-dependent proliferation of rat aortic smooth muscle cells.
abstract: Proliferation of vascular smooth muscle cells plays a key role in the pathogenesis of several disorders of the vascular wall. Endothelin (ET), a vasoactive peptide that signals through a G protein-coupled receptor, has been linked to mitogenesis in vascular smooth muscle cells, but the mechanistic details underlying this activity remain incompletely understood. In the present study, we demonstrate that ET-dependent mitogenesis in rat neonatal and adult aortic smooth muscle (RASM) cells is accompanied by an increase (up to 10-fold) in CDK2 activity, but not CDK2 protein levels. This effect is blocked almost entirely by PD98059 and UO126, implying involvement of the MEK/ERK signal transduction cascade in the activation. Extracts of ET-treated cells phosphorylate the N terminus of WEE1, an inhibitory kinase, which negatively regulates CDK2 activity through phosphorylation at Tyr(15), leading to a decrease in WEE1 activity and a reduction in levels of phospho-Tyr(15) in the CDK2 protein. ET also increases expression and activity of CDC25A, the regulatory phosphatase responsible for dephosphorylating Tyr(15). All of these effects are reversible following treatment with the MEK inhibitor PD98059. ET also increases levels of CDC2 activity in these cells in association with a decrease in levels of phospho-Tyr(15) on the CDC2 molecule. Phosphorylation of WEE1 is linked to ERK while phosphorylation of MYT1 (CDC2-selective inhibitory kinase) is tied to the ribosomal S6 kinase (RSK). In summary, ET controls progression through the cell cycle, in part, by increasing CDK2 and CDC2 activity through the MEK/ERK/RSK signal transduction pathway(s). This results from the phosphorylation and subsequent inactivation of two inhibitory kinases (WEE1 and MYT1) that tonically suppress CDK2 and CDC2 activity and activation of a phosphatase (CDC25A) that increases CDK2 activity.
authors.affiliation: Department of Medicine, University of California at San Francisco, San Francisco, California 94143-0540, USA.
authors.forename: Songcang
authors.initials: S
authors.lastname: Chen
authors.forename: David G
authors.initials: DG
authors.lastname: Gardner
mesh_terms: D000818:Animals; D001011:Aorta / Q000166:cytology; D042846:CDC2-CDC28 Kinases / Q000235:genetics / Q000378:metabolism; D018797:Cell Cycle Proteins; D002455:Cell Division / Q000502:physiology; D051357:Cyclin-Dependent Kinase 2; D016232:Endothelins / Q000378:metabolism*; D004730:Endothelium, Vascular / Q000378:metabolism*; D005260:Female; D020935:MAP Kinase Signaling System / Q000502:physiology; D009131:Muscle, Smooth, Vascular / Q000166:cytology* / Q000378:metabolism; D009687:Nuclear Proteins; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism; D011505:Protein-Tyrosine Kinases / Q000235:genetics / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D020687:cdc25 Phosphatases / Q000378:metabolism*
chemical_list: D018797:Cell Cycle Proteins; D016232:Endothelins; D009687:Nuclear Proteins; C490680:Myt1 kinase, rat; D011505:Protein-Tyrosine Kinases; D017346:Protein Serine-Threonine Kinases; D042846:CDC2-CDC28 Kinases; C495892:Cdk2 protein, rat; D051357:Cyclin-Dependent Kinase 2; C517007:Cdc25a protein, rat; D020687:cdc25 Phosphatases