We shall further develop and apply a 32P-labeling test recently developed by us to detect covalent adducts formed by the reaction of nucleic acids (DNA and, to a lesser extent, RNA) with chemicals. The test involves (i) the reaction of DNA with chemical(s) in vitro or in vivo; (ii) DNA purification; (iii) enzymatic digestion of DNA to deoxynucleoside 3'-monophosphates; (iv) conversion of the latter to 32P-labeled deoxynucleoside 3',5'-bisphosphates by incubation with [Gamma-32P]ATP and T4 polynucleotide kinase; (v) fingerprinting of the 32P-labeled digests by reversed-phase high-performance liquid chromatography and anion-exchange thin-layer chromatorgraphy on polyethyleneimine-cellulose, and (vi) autoradiography. Adducts formed by the covalent binding to DNA of chemical are detected as extra spots on such fingerprints. A similar procedure applies to treatment and analysis of RNA. The test will be applied to carcinogenic alkylating agents, mycotoxins, aminoazo dyes, aromatic amines and amides, polycyclic aromatic hydrocarbons, other aromatics, nitrosamines, chlorinated hydrocarbons, hydrazine derivatives, and antineoplastic agents. A number of noncarcinogens will also be assayed. The results will be compared with published data from mutagenesis and carcinogenesis assays to determine the scope of the test as a prescreen for carcinogens.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
5R37CA032157-10
Application #
3482250
Study Section
Chemical Pathology Study Section (CPA)
Project Start
1982-05-01
Project End
1992-04-30
Budget Start
1991-05-01
Budget End
1992-04-30
Support Year
10
Fiscal Year
1991
Total Cost
Indirect Cost
Name
Baylor College of Medicine
Department
Type
Schools of Medicine
DUNS #
074615394
City
Houston
State
TX
Country
United States
Zip Code
77030
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Randerath, K; Randerath, E; Zhou, G D et al. (1999) Bulky endogenous DNA modifications (I-compounds) -possible structural origins and functional implications. Mutat Res 424:183-94
Zhou, G D; Hernandez, N S; Randerath, E et al. (1999) Acute elevation by short-term dietary restriction or food deprivation of type I I-compound levels in rat liver DNA. Nutr Cancer 35:87-95
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