Document Detail

UVB irradiation-induced activator protein-1 activation correlates with increased c-fos gene expression in a human keratinocyte cell line.
MedLine Citation:
PMID:  9822695     Owner:  NLM     Status:  MEDLINE    
The effects of UVB irradiation on transcription factor activator protein-1 (AP-1) DNA binding and AP-1 transactivation were studied in a human keratinocyte cell line, HaCaT. UVB-induced AP-1 binding to a consensus AP-1 binding site was observed by gel shift assays with maximum stimulation at 12 h after UVB irradiation. A promoter region of the human collagenase-1 gene containing the same AP-1 binding sequence linked to a luciferase reporter gene was stably transfected into HaCaT cells. UVB irradiation significantly increased luciferase activity in these stably transfected cells, with maximum activity observed at 24 h after UVB irradiation. c-Fos and Jun D were identified by antibody clearing assays as the main components of the bound AP-1 complexes. Inhibition of transcription with actinomycin D and inhibition of protein synthesis with cycloheximide significantly abrogated the effect of UVB on AP-1 DNA binding, indicating that transcription and translation were required for AP-1 activation. Northern and Western analyses revealed a correlation between increased AP-1 activity and accumulation of c-fos mRNA and c-Fos protein after UVB irradiation. UVB irradiation increased c-fos transcription in HaCaT cells stably transfected with a plasmid containing the human c-fos promoter driving a luciferase reporter gene. These results suggest that increased c-fos expression may play an important role in UVB-induced AP-1 activation in HaCaT cells.
W Chen; A H Borchers; Z Dong; M B Powell; G T Bowden
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  The Journal of biological chemistry     Volume:  273     ISSN:  0021-9258     ISO Abbreviation:  J. Biol. Chem.     Publication Date:  1998 Nov 
Date Detail:
Created Date:  1998-12-23     Completed Date:  1998-12-23     Revised Date:  2008-11-21    
Medline Journal Info:
Nlm Unique ID:  2985121R     Medline TA:  J Biol Chem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  32176-81     Citation Subset:  IM    
Department of Radiation Oncology, Arizona Cancer Center, College of Medicine, University of Arizona, Tucson, Arizona 85724, USA.
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MeSH Terms
Cell Line
Cycloheximide / pharmacology
Dactinomycin / pharmacology
Dose-Response Relationship, Radiation
Gene Expression Regulation / radiation effects*
Genes, Reporter
Genes, fos / radiation effects*
Keratinocytes / metabolism,  radiation effects*
Luciferases / biosynthesis,  genetics
Proto-Oncogene Proteins c-fos / genetics*
Transcription Factor AP-1 / metabolism,  radiation effects*
Transcription, Genetic / drug effects,  radiation effects
Transcriptional Activation / radiation effects
Ultraviolet Rays*
Grant Support
Reg. No./Substance:
0/Proto-Oncogene Proteins c-fos; 0/Transcription Factor AP-1; 50-76-0/Dactinomycin; 66-81-9/Cycloheximide; EC 1.13.12.-/Luciferases

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