title: Identification of viral genes essential for replication of murine gamma-herpesvirus 68 using signature-tagged mutagenesis.
abstract: Gamma-herpesviruses, Epstein-Barr virus, and Kaposi's sarcoma-associated herpesvirus are important human pathogens, because they are involved in tumor development. Murine gamma-herpesvirus-68 (MHV-68 or gammaHV-68) has emerged as a small animal model system for the study of gamma-herpesvirus pathogenesis and host-virus interactions. To identify the genes required for viral replication in vitro and in vivo, we generated 1,152 mutants using signature-tagged transposon mutagenesis on an infectious bacterial artificial chromosome of MHV-68. Almost every ORF was mutated by random insertion. For each ORF, a mutant with an insertion proximal to the N terminus of each ORF was examined for the ability to grow in fibroblasts. Our results indicate that 41 genes are essential for in vitro growth, whereas 26 are nonessential and 6 attenuated. Replication-competent mutants were pooled to infect mice, which led to the discovery of ORF 54 being important for MHV-68 to replicate in the lung. This genetic analysis of a tumor-associated herpesvirus at the whole genome level validates signature-tagged transposon mutagenesis screening as an effective genetic system to identify important virulent genes in vivo and define interactions with the host immune system.
authors.affiliation: Department of Microbiology, College of Medicine, Hallym University, Chuncheon 200-702, Republic of Korea.
authors.forename: Moon Jung
authors.initials: MJ
authors.lastname: Song
authors.forename: Seungmin
authors.initials: S
authors.lastname: Hwang
authors.forename: Wendy H
authors.initials: WH
authors.lastname: Wong
authors.forename: Ting-Ting
authors.initials: TT
authors.lastname: Wu
authors.forename: Sangmi
authors.initials: S
authors.lastname: Lee
authors.forename: Hsiang-I
authors.initials: HI
authors.lastname: Liao
authors.forename: Ren
authors.initials: R
authors.lastname: Sun
mesh_terms: D000818:Animals; D005814:Genes, Viral / Q000502:physiology*; D051379:Mice; D016296:Mutagenesis; D009154:Mutation; D041681:NIH 3T3 Cells; D016366:Open Reading Frames; D011755:Pyrophosphatases / Q000502:physiology; D018144:Rhadinovirus / Q000235:genetics* / Q000502:physiology; D014779:Virus Replication
chemical_list: D011755:Pyrophosphatases; C020020:dUTP pyrophosphatase
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