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Synthesis of C8‐Arylamine‐Modified 2′‐Deoxyadenosine Phosphoramidites and their Site‐Specific Incorporation into Oligonucleotides
Author(s) -
Szombati Zita,
Baerns Sabrina,
Marx Andreas,
Meier Chris
Publication year - 2012
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201100573
Subject(s) - oligonucleotide , chemistry , dna , ecori , stereochemistry , dna polymerase , polymerase , biochemistry , restriction enzyme
Abstract Adducts of C8‐(N‐acetyl)‐arylamines and 2′‐deoxyadenosine were synthesised by palladium‐catalysed CN 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.