title: Synthesis of C8-arylamine-modified 2'-deoxyadenosine phosphoramidites and their site-specific incorporation into oligonucleotides.
abstract: Adducts of C8-(N-acetyl)-arylamines and 2'-deoxyadenosine were synthesised by palladium-catalysed C--N cross-coupling chemistry. These 2'-dA adducts were converted into the corresponding 3'-phosphoramidites and site-specifically incorporated into DNA oligonucleotides, which were characterised by mass spectrometry, UV thermal-stability assays and circular dichroism. These modified oligonucleotides were also used in EcoRI restriction assays and in primer-extension studies with three different DNA polymerases. The incorporation of the 2'-dA lesion close to the EcoRI restriction site dramatically reduced the susceptibility of the DNA strand to cleavage; this indicates a significant local distortion of the DNA double helix. The incorporation of the acetylated C8-2'-dA-phosphoramidites into 20-mer oligonucleotides failed, however, because the N-acetyl group was lost during the deprotection process. Instead the corresponding C8-NH-2'-dA-modified oligonucleotides were obtained. The effect of the C8-NH-arylamine-dA lesion on the replication by DNA polymerases was clearly dependent both on the polymerase used and on the arylamine-dA damage.
authors.affiliation: Organic Chemistry, Department of Chemistry, University of Hamburg, Martin-Luther-King-Platz 6, 20146 Hamburg, Germany.
authors.forename: Zita
authors.initials: Z
authors.lastname: Szombati
authors.forename: Sabrina
authors.initials: S
authors.lastname: Baerns
authors.forename: Andreas
authors.initials: A
authors.lastname: Marx
authors.forename: Chris
authors.initials: C
authors.lastname: Meier
mesh_terms: D000588:Amines / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism; D002942:Circular Dichroism; D004261:DNA Replication; D004259:DNA-Directed DNA Polymerase / Q000737:chemistry / Q000378:metabolism; D003839:Deoxyadenosines / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism; D015394:Molecular Structure; D009841:Oligonucleotides / Q000737:chemistry* / Q000378:metabolism; D009943:Organophosphorus Compounds / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism; D013696:Temperature
chemical_list: D000588:Amines; D003839:Deoxyadenosines; D009841:Oligonucleotides; D009943:Organophosphorus Compounds; C434331:phosphoramidite; D004259:DNA-Directed DNA Polymerase; C058118:2'-deoxyadenosine