title: Rta of murine gammaherpesvirus 68 reactivates the complete lytic cycle from latency.
abstract: Herpesviruses are characterized as having two distinct life cycle phases: lytic replication and latency. The mechanisms of latency establishment and maintenance, as well as the switch from latency to lytic replication, are poorly understood. Human gammaherpesviruses, including Epstein-Barr virus (EBV) and human herpesvirus-8 (HHV-8), also known as Kaposi's sarcoma-associated herpesvirus (KSHV), are associated with lymphoproliferative diseases and several human tumors. Unfortunately, the lack of cell lines to support efficient de novo productive infection and restricted host ranges of EBV and HHV-8 make it difficult to explore certain important biological questions. Murine gammaherpesvirus 68 (MHV-68, or gammaHV68) can establish de novo lytic infection in a variety of cell lines and is also able to infect laboratory mice, offering an ideal model with which to study various aspects of gammaherpesvirus infection. Here we describe in vitro studies of the mechanisms of the switch from latency to lytic replication of MHV-68. An MHV-68 gene, rta (replication and transcription activator), encoded primarily by open reading frame 50 (ORF50), is homologous to the rta genes of other gammaherpesviruses, including HHV-8 and EBV. HHV-8 and EBV Rta have been shown to play central roles in viral reactivation from latency. We first studied the kinetics of MHV-68 rta gene transcription during de novo lytic infection. MHV-68 rta was predominantly expressed as a 2-kb immediate-early transcript. Sequence analysis of MHV-68 rta cDNA revealed that an 866-nucleotide intron 5' of ORF50 was removed to create the Rta ORF of 583 amino acids. To test the functions of MHV-68 Rta in reactivation, a plasmid expressing Rta was transfected into a latently infected cell line, S11E, which was established from a B-cell lymphoma in an MHV-68-infected mouse. Rta induced expression of viral early and late genes, lytic replication of viral DNA, and production of infectious viral particles. We conclude that Rta alone is able to disrupt latency, activate viral lytic replication, and drive the lytic cycle to completion. This study indicates that MHV-68 provides a valuable model for investigating regulation of the balance between latency and lytic replication in vitro and in vivo.
authors.affiliation: Department of Molecular Pharmacology, the UCLA AIDS Institute, University of California at Los Angeles, Los Angeles, California 90095, USA.
authors.forename: T T
authors.initials: TT
authors.lastname: Wu
authors.forename: E J
authors.initials: EJ
authors.lastname: Usherwood
authors.forename: J P
authors.initials: JP
authors.lastname: Stewart
authors.forename: A A
authors.initials: AA
authors.lastname: Nash
authors.forename: R
authors.initials: R
authors.lastname: Sun
mesh_terms: D000818:Animals; D001402:B-Lymphocytes; D002461:Cell Line, Transformed; D002478:Cells, Cultured; D018142:Gammaherpesvirinae / Q000235:genetics / Q000502:physiology*; D015967:Gene Expression Regulation, Viral; D017874:Immediate-Early Proteins / Q000235:genetics / Q000502:physiology; D051379:Mice; D015534:Trans-Activators / Q000235:genetics / Q000502:physiology*; D014158:Transcription, Genetic; D014162:Transfection; D010948:Viral Plaque Assay; D014764:Viral Proteins / Q000235:genetics / Q000502:physiology*; D014771:Virion / Q000472:pathogenicity; D014775:Virus Activation; D017735:Virus Latency; D014779:Virus Replication
chemical_list: D017874:Immediate-Early Proteins; C116742:ORF 50 transactivator; D015534:Trans-Activators; D014764:Viral Proteins