Characterization of antibodies to dihydrothymine, a radiolysis product of DNA

Journal Article · Tue May 16 04:00:00 EDT 1989 · Biochemistry; (USA)

DOI:https://doi.org/10.1021/bi00436a039· OSTI ID:5348841

Hubbard, K; Ide, H; Erlanger, B F; Wallace, S S ^[1]

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1. New York Medical College, Valhalla (USA)

Antibodies to dihydrothymine were elicited by immunizing rabbits with dihydrothymidine monophosphate conjugated by carbodiimide to BSA. By use of an ELISA assay, the antibodies produced were found to be specific for dihydrothymine. Hapten inhibition studies showed that dihydrothymidine monophosphate was 3 orders of magnitude more effective as an inhibitor than thymidine monophosphate and 4 orders of magnitude more effective than thymidine glycol monophosphate. With DNA containing dihydrothymine, antibody reactivity was observed at 20 fmol of dihydrothymine, which is approximately 0.1 dihydrothymine per 10,000 bases. Thus, the assay is very sensitive. The antibody reacted with denatured DNA containing dihydrothymine but not with native DNA containing this lesion. The antibody was used for measurement of in vivo incorporation of dihydrothymidine in wild-type Escherichia coli or mutants defective in their ability to remove dihydrothymine from DNA or in the de novo synthesis of thymidylate. Lastly, antibodies to dihydrothymine were use to quantitate the formation of dihydrothymine in DNA X-irradiated under N2. Production of dihydrothymine in irradiated DNA correlated with the level of reducing species produced by X-rays, and dihydrothymine was produced preferentially in irradiated single-stranded or denatured DNA as compared to irradiated duplex DNA.

• OSTI ID: 5348841
• Journal Information: Biochemistry; (USA), Journal Name: Biochemistry; (USA) Vol. 28:10; ISSN 0006-2960; ISSN BICHA
• Country of Publication: United States
• Language: English

Related Subjects

560120* -- Radiation Effects on Biochemicals
Cells
& Tissue Culture
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
ANTIBODIES
AZINES
BACTERIA
BIOASSAY
BIOSYNTHESIS
CHEMICAL COMPOSITION
CHEMICAL RADIATION EFFECTS
CHEMISTRY
DNA
ELECTROMAGNETIC RADIATION
ENZYME IMMUNOASSAY
ESCHERICHIA COLI
HETEROCYCLIC COMPOUNDS
HYDROXY COMPOUNDS
IMMUNOASSAY
IONIZING RADIATIONS
MICROORGANISMS
NUCLEIC ACIDS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PYRIMIDINES
RADIATION CHEMISTRY
RADIATION EFFECTS
RADIATIONS
SPECIFICITY
SYNTHESIS
THYMINE
URACILS
X RADIATION