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U.S. Department of Energy
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The impact of energy-related pollutants on chromosome structure: Progress report, May 1, 1988--April 30, 1989

Technical Report ·
OSTI ID:6540135

We have examined, at the DNA sequence level, the sequence preferences of binding of six chemicals to linear DNA. The chemicals utilized were the bis-(1,10)-phenanthroline Cu(I) complex ((Phen)/sub 2/Cu/sup +/), (+)-anti-and (/minus/)-anti isomers of benzo(a)pyrene-7,8-diol-9, 10-epoxide (BPDE), acetylacetoxyamino-fluorene (AAAF), 8-azido-ethidium (AEt), and 7-azido-actinomycin D (AAct D). Mapping of binding sites depends on the abilities of these chemicals to either spontaneously cleave DNA, as in the case of (Phen)/sub 2/Cu/sup +/, or to create alkali-labile base adducts, as in the cases of the other chemicals. The techniques utilized are analogous to those of DNA sequencing. Binding sites were mapped in sea urchin and mouse histone genes cloned into pUC plasmids. Mapping has now been performed on 1000--2000 bases for all of the chemicals excepting AAAF. All six chemical exhibit distinct sequence preferences. Preferences are notable at the level of doublets, but in all cases the preferences are strongest at the levels of triplets or quartets. Some evidence has been obtained for preferences that extend beyond quartets. The most preferred sequences are different for all five chemicals examined in detail. Perhaps most interesting are the differences in specifications noted for the anti-(+) and anti-(/minus/) isomers of BPDE.

Research Organization:
Florida State Univ., Tallahassee (USA). Inst. of Molecular Biophysics
DOE Contract Number:
FG05-87ER60588
OSTI ID:
6540135
Report Number(s):
DOE/ER/60588-2; ON: DE89005269
Country of Publication:
United States
Language:
English