title: Repair synthesis step involving ERCC1-XPF participates in DNA repair of the Top1-DNA damage complex.
abstract: Topoisomerase 1 (Top1) is the intercellular target of camptothecins (CPTs). CPT blocks DNA religation in the Top1-DNA complex and induces Top1-attached nick DNA lesions. In this study, we demonstrate that excision repair cross complementing 1 protein-xeroderma pigmentosum group F (ERCC1-XPF) endonuclease and replication protein A (RPA) participate in the repair of Top1-attached nick DNA lesions together with other nucleotide excision repair (NER) factors. ERCC1-XPF shows nuclease activity in the presence of RPA on a 3'-phosphotyrosyl bond nick-containing DNA (Tyr-nick DNA) substrate, which mimics a Top1-attached nick DNA lesion. In addition, ERCC1-XPF and RPA form a DNA/protein complex on the nick DNA substrate in vitro, and co-localize in CPT-treated cells in vivo. Moreover, the DNA repair synthesis of Tyr-nick DNA lesions occurred in the presence of NER factors, including ERCC1-XPF, RPA, DNA polymerase delta, flap endonuclease 1 and DNA ligase 1. Therefore, some of the NER repair machinery might be an alternative repair pathway for Top1-attached nick DNA lesions. Clinically, these data provide insights into the potential of ERCC1 as a biomarker during CPT regimens.
authors.affiliation: Graduate School of Engineering Science, Osaka University Graduate School of Engineering Science, 1-3 Machikaneyama, Toyonaka, Osaka 560-8531, Japan, Research Institute of Environmental Medicine, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8601, Japan, Department of Toxicogenomics, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya 466-8550, Japan and Laboratories for Organismal Biosystems, Graduate School of Frontier Biosciences, Osaka University, 1-3 Yamadaoka, Suita, Osaka 565-0871, Japan Present address: Centre de Recherche en Cancérologie de Marseille, CRCM/UMR7258, 27, BD Leï Roure BP 30059, 13273 Marseille Cedex 09, France.
authors.forename: Chiaki
authors.initials: C
authors.lastname: Takahata
authors.affiliation: Research Institute of Environmental Medicine, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8601, Japan, Department of Toxicogenomics, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya 466-8550, Japan and.
authors.forename: Yuji
authors.initials: Y
authors.lastname: Masuda
authors.affiliation: Laboratories for Organismal Biosystems, Graduate School of Frontier Biosciences, Osaka University, 1-3 Yamadaoka, Suita, Osaka 565-0871, Japan Present address: Centre de Recherche en Cancérologie de Marseille, CRCM/UMR7258, 27, BD Leï Roure BP 30059, 13273 Marseille Cedex 09, France.
authors.forename: Arato
authors.initials: A
authors.lastname: Takedachi
authors.affiliation: Laboratories for Organismal Biosystems, Graduate School of Frontier Biosciences, Osaka University, 1-3 Yamadaoka, Suita, Osaka 565-0871, Japan.
authors.forename: Kiyoji
authors.initials: K
authors.lastname: Tanaka
authors.affiliation: Graduate School of Engineering Science, Osaka University Graduate School of Engineering Science, 1-3 Machikaneyama, Toyonaka, Osaka 560-8531, Japan, Research Institute of Environmental Medicine, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8601, Japan, Department of Toxicogenomics, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya 466-8550, Japan and Laboratories for Organismal Biosystems, Graduate School of Frontier Biosciences, Osaka University, 1-3 Yamadaoka, Suita, Osaka 565-0871, Japan Present address: Centre de Recherche en Cancérologie de Marseille, CRCM/UMR7258, 27, BD Leï Roure BP 30059, 13273 Marseille Cedex 09, France.
authors.forename: Shigenori
authors.initials: S
authors.lastname: Iwai
authors.affiliation: Graduate School of Engineering Science, Osaka University Graduate School of Engineering Science, 1-3 Machikaneyama, Toyonaka, Osaka 560-8531, Japan, Research Institute of Environmental Medicine, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8601, Japan, Department of Toxicogenomics, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya 466-8550, Japan and Laboratories for Organismal Biosystems, Graduate School of Frontier Biosciences, Osaka University, 1-3 Yamadaoka, Suita, Osaka 565-0871, Japan Present address: Centre de Recherche en Cancérologie de Marseille, CRCM/UMR7258, 27, BD Leï Roure BP 30059, 13273 Marseille Cedex 09, France. Kuraoka@chem.es.osaka-u.ac.jp.
authors.forename: Isao
authors.initials: I
authors.lastname: Kuraoka
mesh_terms: D002166:Camptothecin / Q000494:pharmacology; D004247:DNA / Q000096:biosynthesis; D053904:DNA Breaks, Single-Stranded; D004249:DNA Damage / Q000187:drug effects / Q000502:physiology*; D000072481:DNA Ligase ATP; D011088:DNA Ligases / Q000378:metabolism; D004260:DNA Repair / Q000502:physiology*; D004264:DNA Topoisomerases, Type I / Q000235:genetics / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D004720:Endonucleases / Q000235:genetics / Q000378:metabolism*; D045585:Flap Endonucleases / Q000378:metabolism; D006367:HeLa Cells / Q000187:drug effects; D006801:Humans; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D051716:Replication Protein A / Q000235:genetics / Q000378:metabolism; D014443:Tyrosine / Q000378:metabolism
chemical_list: D004268:DNA-Binding Proteins; D011994:Recombinant Proteins; D051716:Replication Protein A; C095888:xeroderma pigmentosum group F protein; D014443:Tyrosine; D004247:DNA; C071447:ERCC1 protein, human; D004720:Endonucleases; D045585:Flap Endonucleases; C468317:FEN1 protein, human; D004264:DNA Topoisomerases, Type I; C528909:TOP1 protein, human; D011088:DNA Ligases; D000072481:DNA Ligase ATP; D002166:Camptothecin