title: Presence of a trs-Like Motif Promotes Rep-Mediated Wild-Type Adeno-Associated Virus Type 2 Integration.
abstract: High-throughput integration site (IS) analysis of wild-type adeno-associated virus type 2 (wtAAV2) in human dermal fibroblasts (HDFs) and HeLa cells revealed that juxtaposition of a Rep binding site (RBS) and terminal resolution site (trs)-like motif leads to a 4-fold-increased probability of wtAAV integration. Electrophoretic mobility shift assays (EMSAs) confirmed binding of Rep to off-target RBSs. For the first time, we show Rep protein off-target nicking activity, highlighting the importance of the nicking substrate for Rep-mediated integration.
authors.affiliation: Department of Translational Oncology, National Center for Tumor Diseases (NCT) and German Cancer Research Center (DKFZ), Heidelberg, Germany.
authors.forename: Karl
authors.initials: K
authors.lastname: Petri
authors.affiliation: Department of Translational Oncology, National Center for Tumor Diseases (NCT) and German Cancer Research Center (DKFZ), Heidelberg, Germany.
authors.forename: Richard
authors.initials: R
authors.lastname: Gabriel
authors.affiliation: Department of Infectious Diseases, King's College, London, United Kingdom.
authors.forename: Leticia
authors.initials: L
authors.lastname: Agundez
authors.affiliation: Department of Translational Oncology, National Center for Tumor Diseases (NCT) and German Cancer Research Center (DKFZ), Heidelberg, Germany.
authors.forename: Raffaele
authors.initials: R
authors.lastname: Fronza
authors.affiliation: Department of Translational Oncology, National Center for Tumor Diseases (NCT) and German Cancer Research Center (DKFZ), Heidelberg, Germany.
authors.forename: Saira
authors.initials: S
authors.lastname: Afzal
authors.affiliation: Department of Translational Oncology, National Center for Tumor Diseases (NCT) and German Cancer Research Center (DKFZ), Heidelberg, Germany.
authors.forename: Christine
authors.initials: C
authors.lastname: Kaeppel
authors.affiliation: Department of Infectious Diseases, King's College, London, United Kingdom.
authors.forename: R Michael
authors.initials: RM
authors.lastname: Linden
authors.affiliation: Department of Infectious Diseases, King's College, London, United Kingdom els.henckaerts@kcl.ac.uk manfred.schmidt@nct-heidelberg.de.
authors.forename: Els
authors.initials: E
authors.lastname: Henckaerts
authors.affiliation: Department of Translational Oncology, National Center for Tumor Diseases (NCT) and German Cancer Research Center (DKFZ), Heidelberg, Germany els.henckaerts@kcl.ac.uk manfred.schmidt@nct-heidelberg.de.
authors.forename: Manfred
authors.initials: M
authors.lastname: Schmidt
mesh_terms: D020816:Amino Acid Motifs; D002460:Cell Line; D004268:DNA-Binding Proteins / Q000378:metabolism*; D000229:Dependovirus / Q000502:physiology*; D004847:Epithelial Cells / Q000821:virology; D005347:Fibroblasts / Q000821:virology; D006801:Humans; D014764:Viral Proteins / Q000378:metabolism*; D016662:Virus Integration
chemical_list: D004268:DNA-Binding Proteins; D014764:Viral Proteins; C068734:rep proteins, Adeno-associated virus 2