N-2-AMINOFLUORENE AND N-2 ACETYLAMINOFLUORENE ADDUCTS - THE LOCAL SEQUENCE CONTEXT OF AN ADDUCT AND ITS CHEMICAL-STRUCTURE DETERMINE ITS REPLICATION PROPERTIES

作者：BELGUISEVALLADIER P^*; FUCHS RPP

来源：Journal of Molecular Biology, 1995, 249(5): 903-913.

DOI：10.1006/jmbi.1995.0347

• N-2-ACETYLAMINOFLUORENE
• DNA POLYMERASE
• LESION BYPASS
• CHEMICAL PROBES
• STRUCTURAL DEFORMATION

The strong rat liver carcinogen, N-2-acetylaminofluorene, forms mainly two types of guanine adducts at the C-8 position, the acetylaminofluorene adduct (dGuo-C8-AAF) and the aminofluorene adduct (dGuo-C8-AF). We have constructed different oligonucleotides bearing a single AF lesion at each of the guanine residues of the NarI mutagenesis hot spot (G(1)G(2)CG(3)CC) and analysed the structural distortion induced by this DNA adduct according to the sequence context. At position G(1) and G(2), the deformation induced by the AF adduct is smaller than the deformation induced by the corresponding acetylated form of this adduct (i.e. the AAF adduct at the G(1) and G(2)), whereas both AF and AAF adducts induce a similar structural change when bound to G(3). Single-stranded oligonucleotides modified with AF adducts were used in primer extension replication assays using purified DNA polymerases (PolIII holoenzyme, Klenow fragment(exo+ and exo-), Sequenase 2.0) and the data compared to the AAF containing substrates. Translesion synthesis (complete bypass) is found with all tested polymerases when AF adducts are bound to G(1) or G(2) while little or no bypass is seen when the AF adduct is bound to G(3). On the other hand, irrespective of its position within the NarI sequence, AAF adducts completely block DNA synthesis. The results described in this paper show that the sole knowledge of the chemical structure of an adduct neither determines uniquely the conformational change it induces at the DNA level nor its replication properties. Indeed, although AF adducts are in most cases non-distorting adducts and as a consequence non-replication blocking lesions (as exemplified by adducts at G(1) or G(2)), some AF adducts (as at position G(3)) behave almost as AAF adducts in terms of the structural distortion induced and its replication blocking property These findings stress the strong modulation by the local sequence context of the structural and biological consequences of a given adduct.

• 出版日期1995-6-23