Abstract
The cell cycle-dependent transcription factor, E2F-1, regulates the cyclin-like species of the DNA repair enzyme uracil-DNA glycosylase (UDG) gene in human osteosarcoma (Saos-2) cells. We demonstrate, through the deletion of the human UDG promoter sequences, that expression of E2F-1 activates the UDG promoter through several E2F sites. The major putative downstream site for E2F, located in the first exon, serves as a target for E2F-1/DP1 complex binding in vitro. We also provide evidence for the functional relationship between the cyclin-like UDG gene product and E2F. High levels of UDG expression in a transient transfection assay result in the down-regulation of transcriptional activity through elements specific for E2F-mediated transcription. Overexpression of UDG in Saos 2 cells was observed to delay growth late in G1 phase and transiently arrest these cells from progressing into the S phase. This hypothetical model integrates one mechanism of DNA repair with the cell cycle control of gene transcription, likely through E2F. This implicates E2F as a multifunctional target for proteins and enzymes, possibly, responsive to DNA damage through the negative effect of UDG on E2F-mediated transcriptional activity.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Antigens, CD / biosynthesis
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Antigens, CD19
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Antigens, Differentiation, B-Lymphocyte / biosynthesis
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B-Lymphocytes / immunology
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Base Sequence
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Bone Neoplasms
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Carrier Proteins*
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Cell Cycle Proteins*
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Cell Cycle*
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Cell Division
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Cell Line
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Chloramphenicol O-Acetyltransferase / biosynthesis
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DNA Glycosylases*
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DNA Primers
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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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Gene Expression Regulation, Enzymologic*
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Gene Expression Regulation, Neoplastic
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Homeostasis
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Humans
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Molecular Sequence Data
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Mutagenesis, Site-Directed
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N-Glycosyl Hydrolases / biosynthesis*
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Osteosarcoma
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Polymerase Chain Reaction
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Promoter Regions, Genetic*
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Recombinant Proteins / biosynthesis
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Restriction Mapping
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Retinoblastoma-Binding Protein 1
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S Phase
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Transcription Factor DP1
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Transcription Factors / biosynthesis
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Transcription Factors / metabolism*
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Transcription, Genetic*
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Transfection
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Tumor Cells, Cultured
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Uracil-DNA Glycosidase
Substances
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Antigens, CD
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Antigens, CD19
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Antigens, Differentiation, B-Lymphocyte
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Carrier Proteins
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Cell Cycle Proteins
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DNA Primers
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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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Recombinant Proteins
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Retinoblastoma-Binding Protein 1
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TFDP1 protein, human
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Transcription Factor DP1
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Transcription Factors
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Chloramphenicol O-Acetyltransferase
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DNA Glycosylases
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N-Glycosyl Hydrolases
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Uracil-DNA Glycosidase