title: FF483-484 motif of human Polη mediates its interaction with the POLD2 subunit of Polδ and contributes to DNA damage tolerance.
abstract: Switching between replicative and translesion synthesis (TLS) DNA polymerases are crucial events for the completion of genomic DNA synthesis when the replication machinery encounters lesions in the DNA template. In eukaryotes, the translesional DNA polymerase η (Polη) plays a central role for accurate bypass of cyclobutane pyrimidine dimers, the predominant DNA lesions induced by ultraviolet irradiation. Polη deficiency is responsible for a variant form of the Xeroderma pigmentosum (XPV) syndrome, characterized by a predisposition to skin cancer. Here, we show that the FF483-484 amino acids in the human Polη (designated F1 motif) are necessary for the interaction of this TLS polymerase with POLD2, the B subunit of the replicative DNA polymerase δ, both in vitro and in vivo. Mutating this motif impairs Polη function in the bypass of both an N-2-acetylaminofluorene adduct and a TT-CPD lesion in cellular extracts. By complementing XPV cells with different forms of Polη, we show that the F1 motif contributes to the progression of DNA synthesis and to the cell survival after UV irradiation. We propose that the integrity of the F1 motif of Polη, necessary for the Polη/POLD2 interaction, is required for the establishment of an efficient TLS complex.
authors: Biotechnologie et Signalisation Cellulaire, Université de Strasbourg, UMR7242, Illkirch 67412, France.
authors: Nadège
authors: N
authors: Baldeck
authors: Biotechnologie et Signalisation Cellulaire, Université de Strasbourg, UMR7242, Illkirch 67412, France.
authors: Régine
authors: R
authors: Janel-Bintz
authors: Biotechnologie et Signalisation Cellulaire, Université de Strasbourg, UMR7242, Illkirch 67412, France.
authors: Jérome
authors: J
authors: Wagner
authors: UMR-S1052, Inserm, Centre de Recherche en Cancérologie de Lyon, Lyon 69000, France.
authors: Agnès
authors: A
authors: Tissier
authors: Cancer Research Center of Marseille (CRCM), Centre National de la Recherche Scientifique, Unité Mixte de Recherche 7258, Marseille 13009, France.
authors: Robert P
authors: RP
authors: Fuchs
authors: Institute of Genetics, Biological Research Center, Hungarian Academy of Sciences, HU-6726 Szeged, Hungary.
authors: Peter
authors: P
authors: Burkovics
authors: Institute of Genetics, Biological Research Center, Hungarian Academy of Sciences, HU-6726 Szeged, Hungary.
authors: Lajos
authors: L
authors: Haracska
authors: Université Paris-Sud, CNRS-UMR8200, Equipe labellisée Ligue Contre le Cancer, Gustave Roussy, Villejuif, France.
authors: Emmanuelle
authors: E
authors: Despras
authors: Biotechnologie et Signalisation Cellulaire, Université de Strasbourg, UMR7242, Illkirch 67412, France.
authors: Marc
authors: M
authors: Bichara
authors: Biotechnologie et Signalisation Cellulaire, Université de Strasbourg, UMR7242, Illkirch 67412, France.
authors: Bruno
authors: B
authors: Chatton
authors: Biotechnologie et Signalisation Cellulaire, Université de Strasbourg, UMR7242, Illkirch 67412, France agnes.cordonnier@unistra.fr.
authors: Agnès M
authors: AM
authors: Cordonnier
mesh_terms: D002453:Cell Cycle; D002460:Cell Line; D002470:Cell Survival; D004249:DNA Damage; D004258:DNA Polymerase III / Q000378:metabolism*; D004259:DNA-Directed DNA Polymerase / Q000737:chemistry* / Q000378:metabolism*; D006801:Humans; D054730:Protein Interaction Domains and Motifs; D021122:Protein Subunits / Q000378:metabolism; D014466:Ultraviolet Rays; D000099270:Y-Family DNA Polymerases
chemical_list: D004258:DNA Polymerase III; D004259:DNA-Directed DNA Polymerase; D021122:Protein Subunits; C000598192:POLD2 protein, human; C112338:Rad30 protein; D000099270:Y-Family DNA Polymerases