Molecular biology of environmental aromatic hydrocarbons. Progress report, September 1, 1984-June 31, 1985

Technical Report · Mon Jul 01 04:00:00 EDT 1985

OSTI ID:5484065

Weiss, S B

The biological activities of the (+)- and (-)-enantiomers of anti-BPDE (benzo(a)pyrene diol epoxide) and BePe (benzo(e)pyrene epoxide) were examined for their capacity to inhibit infectious single- and double-stranded 0X174 phage DNAs. For both activated PAH derivatives, the (+)-isomer was more inhibitory using either single- or double-stranded 0X DNAs. Both PAH derivatives showed a higher inhibition potency with single-stranded 0X DNA than with duplex DNA; this difference between the two phage DNA forms was much greater for BePE than with anti-BPDE. Digestion of phage DNAs reacted with the two isomers of anti-BPDE, followed by chromatography on LH20 Sephadex, showed a single major dG adduct peak for the (+)-isomer suggesting that alkylation of both 0X DNA forms is highly stereoselective. Reaction of the (-)-isomer of anti-BPDE with either form of 0X DNA showed several dG adduct peaks indicating that adduct formation was not stereoselective. A model viral DNA system was used, containing short oligonucleotide inserts as targets for PAH alkylation, to detect sequence modifications induced by anti-BPDE. A 10-base-pair oligomer (Bam HI linker) was treated with anti-BPDE and inserted into phage M13 replicative form DNA. E. coli was transfected with the recombinant DNA containing the alkylated oligomer, progeny viral plaques were selected, and their DNAs subjected to DNA sequence analysis at the region of oligomer insertion. For the alkylated inserts used in our study, the DNA sequence analysis of progeny viral DNA showed that nucleotide deletions were present in all the clones examined. These deletions occurred primarily, but not exclusively, at G dot C cluster regions, varied from 1 to 24 base pairs in length, and included both target and nontarget nucleotides. 19 refs., 4 figs.

• Research Organization: Chicago Univ., IL (USA)
• DOE Contract Number: AC02-80EV10328
• OSTI ID: 5484065
• Report Number(s): DOE/EV/10328-4; ON: DE85015036
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ADDUCTS
AROMATICS
AZAARENES
BACTERIA
BACTERIOPHAGES
BENZOPYRENE
BIOLOGICAL RECOVERY
BIOLOGICAL REPAIR
CHROMATOGRAPHY
CONDENSED AROMATICS
DNA
DNA ADDUCTS
DNA REPAIR
DNA SEQUENCING
EPOXIDES
ESCHERICHIA COLI
GUANOSINE
HETEROCYCLIC COMPOUNDS
HYDROCARBONS
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
LIQUID COLUMN CHROMATOGRAPHY
MICROORGANISMS
NUCLEIC ACIDS
NUCLEOSIDES
NUCLEOTIDES
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
ORGANIC OXYGEN COMPOUNDS
PARASITES
PURINES
RECOMBINANT DNA
RECOVERY
REPAIR
RIBOSIDES
SEPARATION PROCESSES
STRUCTURAL CHEMICAL ANALYSIS
TRACER TECHNIQUES
TRITIUM COMPOUNDS
VIRUSES