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

Chen, L.L.; Clawson, M.L.; Bilgrami, S.; Carmichael, G.

Cell Growth and Differentiation the Molecular Biology Journal of the American Association for Cancer Research 4(12): 975-983

1993


ISSN/ISBN: 1044-9523
PMID: 8117624
Accession: 008075119

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Abstract
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.