title: Rad51 and Rad52 are involved in homologous recombination of replicating herpes simplex virus DNA.
abstract: Replication of herpes simplex virus 1 is coupled to recombination, but the molecular mechanisms underlying this process are poorly characterized. The role of Rad51 and Rad52 recombinases in viral recombination was examined in human fibroblast cells 1BR.3.N (wild type) and in GM16097 with replication defects caused by mutations in DNA ligase I. Intermolecular recombination between viruses, tsS and tsK, harboring genetic markers gave rise to ∼17% recombinants in both cell lines. Knock-down of Rad51 and Rad52 by siRNA reduced production of recombinants to 11% and 5%, respectively, in wild type cells and to 3% and 5%, respectively, in GM16097 cells. The results indicate a specific role for Rad51 and Rad52 in recombination of replicating herpes simplex virus 1 DNA. Mixed infections using clinical isolates with restriction enzyme polymorphisms in the US4 and US7 genes revealed recombination frequencies of 0.7%/kbp in wild type cells and 4%/kbp in GM16097 cells. Finally, tandem repeats in the US7 gene remained stable upon serial passage, indicating a high fidelity of recombination in infected cells.
authors.affiliation: Department of Medical Biochemistry and Cell Biology, Institute of Biomedicine, The Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden.
authors.forename: Ka-Wei
authors.initials: KW
authors.lastname: Tang
authors.affiliation: Department of Infectious Diseases, Section of Virology, Institute of Biomedicine, The Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden.
authors.forename: Peter
authors.initials: P
authors.lastname: Norberg
authors.affiliation: Department of Infectious Diseases, Section of Virology, Institute of Biomedicine, The Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden.
authors.forename: Martin
authors.initials: M
authors.lastname: Holmudden
authors.affiliation: Department of Medical Biochemistry and Cell Biology, Institute of Biomedicine, The Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden.
authors.forename: Per
authors.initials: P
authors.lastname: Elias
authors.affiliation: Department of Infectious Diseases, Section of Virology, Institute of Biomedicine, The Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden.
authors.forename: Jan-Åke
authors.initials: JÅ
authors.lastname: Liljeqvist
mesh_terms: D000818:Animals; D002460:Cell Line; D004261:DNA Replication; D004279:DNA, Viral; D016679:Genome, Viral; D018259:Herpesvirus 1, Human / Q000235:genetics*; D059765:Homologous Recombination; D006801:Humans; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics; D051135:Rad51 Recombinase / Q000235:genetics / Q000378:metabolism*; D051721:Rad52 DNA Repair and Recombination Protein / Q000235:genetics / Q000378:metabolism*
chemical_list: D004279:DNA, Viral; D034741:RNA, Small Interfering; D051721:Rad52 DNA Repair and Recombination Protein; D051135:Rad51 Recombinase