Site-specific introduction of carcinogenic aromatic amines into synthetic oligonucleotides

Conference · Thu May 01 04:00:00 EDT 1986 · Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)

OSTI ID:6997677

Reid, T M; Lee, M S; Gupta, P K; King, C M

Formation of site-specific DNA adducts allows the synthesis of chemically defined lesions that can be used for study of the functional consequences of these reactions. A reactive derivative, N-acetoxy-2-aminofluorene (N-Aco-AF), can be formed by enzymatic O-acetylation of the carcinogen, N-hydroxy-2-aminofluorene. Synthetic N-acetoxy-N-trifluoro-acetyl-2-aminofluorene (N-Aco-N-TFA-AF) has now been shown to undergo solvolysis in ethanolic citrate buffer, losing the trifluoroacetyl moiety to yield the reactive N-Aco-AF. Reaction of N-Aco-N-TFA-AF with guanine-containing mono-, oligo- and polynucleotides yields only N-(guanin-8-yl)-2-aminofluorene derivatives. Although reaction with poly(dA) occurs to a small extent, no reaction occurs with adenine mononucleotides. N-Aco-N-TFA-AF has been reacted with synthetic oligonucleotides, 7 and 17 bases long, that contained only a single guanine residue. HPLC purification, UV spectral analysis and anhydrous trifluoroacetic acid hydrolysis indicates that the modified oligonucleotides contain a single 2-aminofluorene adduct located at the lone guanine. Similar analysis of these oligonucleotides modified with N-acetoxy-N-acetyl-2-aminofluorene shows that they contain only a single 2-acetylaminofluorene adduct, N-(guanin-8-yl)-2-acetylaminofluorene. These modified oligonucleotides are being used to construct viral and plasmid DNA with site-specific lesions.

• Research Organization: Michigan Cancer Foundation, Detroit
• OSTI ID: 6997677
• Report Number(s): CONF-8606151-
• Conference Information: Journal Name: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States) Journal Volume: 45:6
• Country of Publication: United States
• Language: English

Related Subjects

560300* -- Chemicals Metabolism & Toxicology
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
ACETYLATION
ACYLATION
ADDUCTS
AMINES
AROMATICS
CARCINOGENS
CHEMICAL REACTIONS
CHROMATOGRAPHY
CONDENSED AROMATICS
DNA ADDUCTS
FLUORENE
HYDROCARBONS
LIQUID COLUMN CHROMATOGRAPHY
MOLECULAR STRUCTURE
NUCLEOTIDES
ORGANIC COMPOUNDS
SEPARATION PROCESSES