Characterization of the CDP-like/CTAS-1 binding site in the okadaic acid response element (OARE) of the human CDK1(p34cdc2) promoter.
BACKGROUND
Transcription of CDK1 is induced at the G0/G1-phase of the cell cycle and after okadaic acid treatment and we identified the Site I okadaic acid response element (OARE -944 to -763nt) enhancer in the human CDK1 promoter.


METHODS
The OARE region of the CDK1 promoter was characterized for enhancer/repressor activities.


RESULTS
Transient transfection of upper and lower Site I subregions suggested enhanced transcription activity was divided between both while mobility shift assays demonstrated sequence-specific protein binding to Site IA. Site IA also formed shift complexes following serum starvation/refeeding and putative transcription factor binding sites clustered in Site IA. Oligonucleotides encoding a consensus CDP transcription factor binding site effectively competed against authentic Site IA in mobility shift assays. Mutation of the CDP-like binding sequence substantially reduced competition. DNaseI footprinting revealed binding at this site.


CONCLUSIONS
The CDP-like recognition sequence appears to comprise the OARE binding authentic CDP and/or related factors. We termed this site the CDK1 transcription activation sequence-1 (CTAS-1) enhancer of the human CDK1 promoter.
Department of Pathobiology, College of Veterinary Medicine, Auburn University, Auburn, AL 36849-5519, USA. birdric@vetmed.auburn.edu
R Curtis
RC
Bird
Patricia
P
Deinnocentes
D001483:Base Sequence; D001665:Binding Sites; D016203:CDC2 Protein Kinase / Q000096:biosynthesis / Q000235:genetics*; D018983:DNA Footprinting; D003850:Deoxyribonuclease I / Q000378:metabolism; D004586:Electrophoresis; D005786:Gene Expression Regulation / Q000187:drug effects; D006367:HeLa Cells; D006801:Humans; D008387:Marine Toxins; D008969:Molecular Sequence Data; D019319:Okadaic Acid / Q000494:pharmacology*; D010080:Oxazoles / Q000494:pharmacology; D011401:Promoter Regions, Genetic / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D020218:Response Elements; D015533:Transcriptional Activation; D014162:Transfection
D008387:Marine Toxins; D010080:Oxazoles; D012333:RNA, Messenger; D019319:Okadaic Acid; C059041:calyculin A; D016203:CDC2 Protein Kinase; D003850:Deoxyribonuclease I