Locating binding sites for the carcinogen N-acetoxy-N-acetyl-2-aminofluorene using restriction enzyme inhibition assays.
Restriction enzyme inhibition studies have been employed to map the locations of high affinity binding sites of the carcinogen N-acetoxy-N-acetyl-2-aminofluorene (acetoxyAAF) on pBR322, phiX174 and SV40 DNAs. Bound carcinogen levels were kept low (less than 20 bound AAF moieties per DNA molecule) in order to observe only the binding to the high affinity sites. Inhibition of certain restriction enzymes was observed in a limited number of locations on these DNAs. Inhibition increased as bound AAF increased and the particular restriction enzymes inhibited varied with location. On all three DNAs, activities of these enzymes was not affected in other locations. Comparison of the sequences at the sites of inhibition on the three DNAs indicates that all sites have common sequence elements: the presence of either the sequence T(C/G)TT(G/C) or the sequence T(G/C)CTT(G/C).
Mallamaci
M A
MA
Department of Biochemistry, Rutgers University, New Brunswick, NJ 08903.
Bascoy
M L
ML
Brown
J
J
Combates
N J
NJ
Winkle
S A
SA
D000099:Acetoxyacetylaminofluorene / Q000378:metabolism*; D010584:Bacteriophage phi X 174 / Q000235:genetics; D001483:Base Sequence; D001665:Binding Sites; D004249:DNA Damage; D016172:DNA Fingerprinting / Q000379:methods*; D004262:DNA Restriction Enzymes / Q000037:antagonists & inhibitors*; D004269:DNA, Bacterial / Q000378:metabolism*; D004279:DNA, Viral / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics; D008969:Molecular Sequence Data; D013539:Simian virus 40 / Q000235:genetics
D004269:DNA, Bacterial; D004279:DNA, Viral; D000099:Acetoxyacetylaminofluorene; D004262:DNA Restriction Enzymes