title: Regulation of cdk2 activity in endothelial cells that are inhibited from growth by cell contact.
abstract: Endothelial cells (ECs) are quiescent in normal blood vessels but undergo rapid bursts of proliferation after vascular injury and during angiogenesis. Here we show that the activity of cyclin-dependent kinase-2 (cdk2), a key regulator of the G1 and S phases of the cell cycle, is expressed at high levels in proliferating ECs but at low levels in ECs that are contact-inhibited for growth. Despite these differences in kinase activity, the protein levels of cdk2 and 1 of its activating subunits, cyclin E, are not modulated by these different growth conditions. The cdk inhibitor p27 is highly expressed in contact-inhibited but not proliferating ECs, whereas the level of cyclin A protein is preferentially expressed in proliferating ECs. p27 protein was detected in immunoprecipitable complexes with cdk2 or cyclin E in cultures that were contact-inhibited for growth. The functional significance of the p27 induction was indicated by the detection of a heat-stable cdk2 inhibitory activity that was induced by endothelial cell-cell contact and could be immunodepleted with anti-p27 antibodies. In a confluent EC monolayer, cdk2 kinase activity was activated by a scraping injury that led to cell migration and proliferation. The injury-induced activation of cdk2 coincided with the downregulation of p27 and the induction of cyclin A. These data demonstrate that p27 is induced in confluent cultures of ECs. They also indicate that both p27 induction and cyclin A downregulation contribute to the inhibition of cdk2 and cell proliferation by cell-cell contact in ECs.
authors.affiliation: Division of Cardiovascular Research, St. Elizabeth's Medical Center, Tufts University School of Medicine, Boston, MA, USA.
authors.forename: D
authors.initials: D
authors.lastname: Chen
authors.forename: K
authors.initials: K
authors.lastname: Walsh
authors.forename: J
authors.initials: J
authors.lastname: Wang
mesh_terms: D000818:Animals; D000906:Antibodies / Q000494:pharmacology; D001011:Aorta / Q000166:cytology; D042846:CDC2-CDC28 Kinases; D002417:Cattle; D018797:Cell Cycle Proteins; D002455:Cell Division / Q000502:physiology; D002465:Cell Movement / Q000502:physiology; D002478:Cells, Cultured; D003260:Contact Inhibition / Q000502:physiology*; D019925:Cyclin A / Q000378:metabolism; D019927:Cyclin E / Q000378:metabolism; D051357:Cyclin-Dependent Kinase 2; D050760:Cyclin-Dependent Kinase Inhibitor p27; D018844:Cyclin-Dependent Kinases / Q000378:metabolism*; D004730:Endothelium, Vascular / Q000166:cytology* / Q000201:enzymology* / Q000293:injuries; D004789:Enzyme Activation / Q000502:physiology; D006801:Humans; D008869:Microtubule-Associated Proteins / Q000276:immunology / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D013936:Thymidine / Q000378:metabolism / Q000494:pharmacology; D014316:Tritium; D025521:Tumor Suppressor Proteins; D014471:Umbilical Veins / Q000166:cytology; D014945:Wound Healing / Q000502:physiology
chemical_list: D000906:Antibodies; D018797:Cell Cycle Proteins; D019925:Cyclin A; D019927:Cyclin E; D008869:Microtubule-Associated Proteins; D025521:Tumor Suppressor Proteins; D014316:Tritium; D050760:Cyclin-Dependent Kinase Inhibitor p27; D017346:Protein Serine-Threonine Kinases; D042846:CDC2-CDC28 Kinases; C495894:CDK2 protein, human; D051357:Cyclin-Dependent Kinase 2; D018844:Cyclin-Dependent Kinases; D013936:Thymidine