Identification by exonuclease footprinting of a distal promoter-binding protein from HeLa cell extracts.
Incubation of a HeLa cell fraction termed P1000 with genomic DNA fragments containing portions of the 5'-flanking region of the ovalbumin gene, followed by digestion with exonucleases, shows that this cell fraction contains a protein (or proteins) that binds to the distal promoter of the ovalbumin gene. The protein protects both strands and spans a minimum of 22 bp on the noncoding strand from nucleotides -68 to -90. The 5' border of the protected region on the coding strand is located at nucleotide -81. A similar site on the chick beta-globin gene is also protected by HeLa cell fraction P1000. The exonuclease footprinting methods used to identify the distal promoter-binding protein are more sensitive than the DNaseI footprinting method, and can be used to identify sequence-specific binding proteins in a mixture of DNA binding proteins.
Elbrecht
A
A
Tsai
S Y
SY
Tsai
M J
MJ
O'Malley
B W
BW
D001483:Base Sequence; D001665:Binding Sites; D002874:Chromosome Mapping; D004268:DNA-Binding Proteins / Q000235:genetics / Q000302:isolation & purification*; D005092:Exonucleases; D005260:Female; D005914:Globins / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D010047:Ovalbumin / Q000235:genetics; D011401:Promoter Regions, Genetic; D014158:Transcription, Genetic
D004268:DNA-Binding Proteins; D005914:Globins; D010047:Ovalbumin; D005092:Exonucleases