Involvement of ERCC1/XPF and XPG in oligodeoxynucleotide-directed gene modification

Abstract

Oligodeoxynucleotide (ODN)-mediated gene alteration was postulated to occur in two steps, DNA strand pairing and DNA repair. Once alignment has occurred through homologous strand pairing, a single mismatch is formed between an oligonucleotide and one of the target strands. Because of this mismatch, it has been suggested that proteins involved in a mismatch repair pathway (MMR) participate in the process. We proposed an alternative model, in which a transient assimilation of ODN to the target DNA can interrupt the trafficking of RNA polymerase, and the stalled RNA polymerase may signal for recruitment of DNA repair proteins, including transcription-coupled (TCR) DNA repair and nucleotide excision repair (NER) pathways. Recently, we found that transcription of many genes participating in NER and MMR was induced by the presence of plasmid DNA, and the extent of induction correlated with episomal gene repair rates. To investigate whether an increased level of induction of genes involved in specific DNA repair pathways has a functional role in ODN-directed gene repair, we performed episomal targeting in several cell lines with a specific defective gene in NER and MMR pathways. Comparison among several genetically related cell lines harboring a specific defective gene and complementation of missing activities showed that a primary pathway for gene correction involves some of the proteins participating in NER, primarily two endonucleases processing a DNA lesion, but not MMR.