Presence of a trs-Like Motif Promotes Rep-Mediated Wild-Type Adeno-Associated Virus Type 2 Integration

Karl Petri, Richard Gabriel, Leticia Agundez, Raffaele Fronza, Saira Afzal, Christine Kaeppel, R Michael Linden, Els Henckaerts, Manfred Schmidt

Research output: Contribution to journalArticlepeer-review

10 Citations (Scopus)

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.
Original languageEnglish
Pages (from-to)7428-7432
Number of pages5
JournalJournal of Virology
Volume89
Issue number14
Early online date18 May 2015
DOIs
Publication statusPublished - 15 Jul 2015

Keywords

  • Amino Acid Motifs
  • Cell Line
  • DNA-Binding Proteins
  • Dependovirus
  • Epithelial Cells
  • Fibroblasts
  • Humans
  • Viral Proteins
  • Virus Integration

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