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J Biol Chem, Vol. 273, Issue 34, 21594-21602, August 21, 1998
From the Laboratory of Molecular Biology, Division of Basic
Sciences, NCI, National Institutes of Health, Bethesda, Maryland
20892-4255
GC-binding factor (GCF) represses transcription
of certain genes and is encoded by a 3.0-kilobase mRNA (Kageyama,
R., and Pastan, I. (1989) Cell 59, 815-825). The GCF
cDNA hybridizes to two additional mRNA species, 4.2 and 1.2 kilobases. We have used differential hybridization to identify a
cDNA clone (termed GCF2) for the 4.2-kilobase mRNA and find
that it is highly expressed in HUT-102 cells. The open reading frame
consists of 2256 nucleotides and encodes a protein of 752 amino acids
with a calculated molecular mass of 83 kilodaltons. GCF2 expressed
in vitro using reticulocyte lysates and Escherichia
coli migrates as a 160-kilodalton protein in SDS-polyacrylamide
gel electrophoresis but has a molecular mass of 83 kilodaltons as
determined by mass spectrum analysis. GCF2 binds to epidermal growth
factor receptor promoter fragments, and the major binding site is
located between nucleotides
Molecular Cloning and Characterization of a Transcription
Regulator with Homology to GC-binding Factor
249 and 233. Cotransfection assays show
that GCF2 acts to repress transcription from the epidermal growth
factor receptor promoter in constructs containing the major GCF2
binding site and not when the site had been mutated. Thus, GCF2 is a
newly identified transcriptional repressor with aberrant
electrophoretic mobility.
Copyright © 1998 by The American Society for Biochemistry and Molecular Biology, Inc.
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