title: Transcription factor Sp1 mediates p38MAPK-dependent activation of the p21WAF1 gene promoter in vascular smooth muscle cells by pyrrolidine dithiocarbamate.
abstract: Previously, we demonstrated that pyrrolidine dithiocarbamate (PDTC) induced G1 cell cycle arrest in vascular smooth muscle cells (VSMC) through inducing p21WAF1 expression. It has recently been reported that the transcription factors involved in p21WAF1 activation by certain signaling factors may vary in different cell types. However, little is known concerning the molecular mechanisms by which PDTC induces p21WAF1 gene expression in VSMC. In this report, we demonstrate that PDTC induces the p21WAF1 expression at the mRNA level. This increase in p21WAF1 gene expression was due to p38MAPK-dependent activation of the p21WAF1 promoter by PDTC. Transcription factor Sp1 binding site was identified as the cis-element for the activation of p21WAF1 promoter by PDTC, as determined by deletion and mutation analysis. In addition, gel shift and supershift assays demonstrated that this PDTC-responsive element binds specifically to the transcription factor Sp1. Treatment with SB203580, an inhibitor of the p38MAPK, significantly down-regulated transactivation of PDTC-induced Sp1. Finally, the transient expression of VSMC with dominant negative p38MAPK plasmid suppressed PDTC-stimulated Sp1 activity. In conclusion, we report the novel finding that transcription factor Sp1 that is involved in the p38MAPK-mediated control of p21WAF1 regulation on VSMC in response to PDTC has now been identified.
authors.affiliation: National Research Laboratory for Glycobiology, Korean Ministry of Science and Technology, Kyungju, Kyungbuk 780-714, Republic of Korea.
authors.forename: Sung-Kwon
authors.initials: SK
authors.lastname: Moon
authors.forename: Sun-Young
authors.initials: SY
authors.lastname: Jung
authors.forename: Cheorl-Ho
authors.initials: CH
authors.lastname: Kim
mesh_terms: D000975:Antioxidants / Q000494:pharmacology; D001665:Binding Sites; D015152:Blotting, Northern; D002478:Cells, Cultured; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins / Q000235:genetics*; D015536:Down-Regulation; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D016193:G1 Phase / Q000187:drug effects; D017353:Gene Deletion; D015971:Gene Expression Regulation, Enzymologic; D005799:Genes, Dominant; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology; D015151:Immunoblotting; D008156:Luciferases / Q000378:metabolism; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism*; D008957:Models, Genetic; D009131:Muscle, Smooth, Vascular / Q000166:cytology*; D016296:Mutagenesis; D032389:Myocytes, Smooth Muscle / Q000166:cytology; D010957:Plasmids / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D011725:Pyridines / Q000494:pharmacology; D011759:Pyrrolidines / Q000494:pharmacology*; D012313:RNA / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D015398:Signal Transduction; D016329:Sp1 Transcription Factor / Q000502:physiology*; D013859:Thiocarbamates / Q000494:pharmacology*; D015533:Transcriptional Activation; D014162:Transfection; D048051:p38 Mitogen-Activated Protein Kinases
chemical_list: D000975:Antioxidants; C493956:CDKN1A protein, human; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins; D004791:Enzyme Inhibitors; D007093:Imidazoles; D011725:Pyridines; D011759:Pyrrolidines; D012333:RNA, Messenger; D016329:Sp1 Transcription Factor; D013859:Thiocarbamates; C020972:pyrrolidine dithiocarbamic acid; D012313:RNA; D008156:Luciferases; D020928:Mitogen-Activated Protein Kinases; D048051:p38 Mitogen-Activated Protein Kinases; C093642:SB 203580