Premium
Long‐patch DNA repair synthesis during base excision repair in mammalian cells
Author(s) -
Sattler Ulrike,
Frit Philippe,
Salles Bernard,
Calsou Patrick
Publication year - 2003
Publication title -
embo reports
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 4.584
H-Index - 184
eISSN - 1469-3178
pISSN - 1469-221X
DOI - 10.1038/sj.embor.embor796
Subject(s) - dna repair , nucleotide excision repair , base excision repair , dna , microbiology and biotechnology , biology , genetics , computational biology
The base excision repair (BER) process removes base damage such as oxidation, alkylation or abasic sites. Two BER sub‐pathways have been characterized using in vitro methods, and have been classified according to the length of the repair patch as either ‘short‐patch’ BER (one nucleotide) or ‘long‐patch’ BER (LP‐BER; more than one nucleotide). To investigate the occurrence of LP‐BER in vivo , we developed an assay using a plasmid containing a single modified base in the transcribed strand of the enhanced green fluorescent protein (EGFP) gene and a stop codon, based on a single‐nucleotide mismatch, at varying distances on the 3′ side of the lesion. The reversion of the stop codon occurs after DNA repair synthesis and restores EGFP expression after transfection of mismatch‐repair‐deficient cells. Repair patches longer than one nucleotide were observed for 55–80% or 80–100% of the plasmids with a mean length of 2–6 or 6–12 nucleotides for 8‐oxo‐7,8‐dihydroguanine or a synthetic abasic site, respectively. These data show the existence of LP‐BER in vivo , and emphasize the effect of the type of BER substrate lesion on both the yield and the extent of the LP‐BER sub‐pathway.