A sequence-specific single-stranded DNA-binding protein that is responsive to epidermal growth factor recognizes an S1 nuclease-sensitive region in the epidermal growth factor receptor promoter.
An epidermal growth factor (EGF) responsive DNA-binding protein (ERDBP-1) has been identified. It recognizes with high affinity and specificity a specific single-stranded DNA sequence located in the S1 nuclease-sensitive site of the EGF receptor (EGFR) 5' flanking region. The EGF-responsive element, determined by footprint analysis, is located from -364 to -344 (86-106 base pairs upstream from the major in vivo transcription initiation site). The factor does not recognize the antisense DNA sequence or double-stranded DNA of the EGF-responsive element. Three bands were observed by mobility shift assay using nuclear extracts from normal human keratinocytes. UV cross-linking followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed one major band with molecular weight in the range of 121,000 to 128,000. The induction of ERDBP-1 became evident 3 to 4 h after EGF stimulation and remained elevated as long as EGF was present. HL60 cells are devoid of endogenous EGFR and produce no ERDBP-1. Retroviral gene transfer of EGFR into HL60 cells resulted in induction of ERDBP-1 by EGF to levels comparable to those found in human keratinocytes.
Chen
L L
LL
Department of Medicine, University of Connecticut Health Center, Farmington 06030.
Clawson
M L
ML
Bilgrami
S
S
Carmichael
G
G
D001483:Base Sequence; D001665:Binding Sites; D002478:Cells, Cultured; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004815:Epidermal Growth Factor / Q000502:physiology*; D066246:ErbB Receptors / Q000235:genetics*; D006801:Humans; D015603:Keratinocytes / Q000378:metabolism; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D015719:Single-Strand Specific DNA and RNA Endonucleases / Q000378:metabolism; D015533:Transcriptional Activation; D014466:Ultraviolet Rays
D004268:DNA-Binding Proteins; D004815:Epidermal Growth Factor; D066246:ErbB Receptors; D015719:Single-Strand Specific DNA and RNA Endonucleases