Abstract
The synthesis of proliferating cell nuclear antigen (PCNA) is strictly regulated during the cell cycle. To investigate the contribution of the promoter region to the up-regulation of human PCNA expression at the onset of S phase, we have examined 17 putative elements with reporter assays in quiescent L-O2 cells and following serum stimulation. The E2F-like sequence 5'-TTCCCCGCAA-3' located at -84 to -75 is required for the serum-induced transactivation. In electrophoretic mobility shift assays, nuclear extracts from asynchronous L-O2 cells exhibit two binding activities toward the -75 E2F oligonucleotide, and the minor band, whose formation could be interfered with by E2F-1 antibody, represents an S phase-specific complex. This is the first demonstration of the E2F site in the human PCNA 5' promoter as a serum-responsive element.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Cell Cycle Proteins*
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Cell Division
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Cell Line
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Cell Nucleus / metabolism
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Cell Separation
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Cloning, Molecular
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DNA-Binding Proteins*
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E2F Transcription Factors
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E2F1 Transcription Factor
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Flow Cytometry
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G1 Phase
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Humans
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Luciferases / metabolism
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Plasmids / metabolism
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Proliferating Cell Nuclear Antigen / blood
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Proliferating Cell Nuclear Antigen / genetics*
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Proliferating Cell Nuclear Antigen / metabolism
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Promoter Regions, Genetic
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Protein Binding
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Resting Phase, Cell Cycle
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Time Factors
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Transcription Factors / blood
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Transcription Factors / chemistry*
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Transcription Factors / metabolism
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Transfection
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Up-Regulation*
Substances
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Cell Cycle Proteins
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DNA-Binding Proteins
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E2F Transcription Factors
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E2F1 Transcription Factor
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E2F1 protein, human
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Proliferating Cell Nuclear Antigen
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Transcription Factors
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Luciferases