title: Mutations in the 5' end of the herpes simplex virus type 2 latency-associated transcript (LAT) promoter affect LAT expression in vivo but not the rate of spontaneous reactivation of genital herpes.
abstract: The primary herpes simplex virus type 2 (HSV-2) latency-associated transcript (LAT) promoter influences LAT expression and rates of virus reactivation. We explored the biological importance of particular neuronally responsive regions within the promoter by creating new recombinant viruses bearing a targeted deletion (246 bp [strain 524]) or a point mutation (2 bp [strain 167]) in this region. These recombinant viruses grew efficiently in vitro and in vivo, caused acute genital disease in guinea pigs, and, as measured by quantitative-competitive (QC) DNA PCR, established latency, all as well as did the wild-type parental HSV-2 strain 333, the rescuant strain 524R, and the previously described 624-bp LAT- promoter deletion mutant. By QC-reverse transcriptase PCR of RNA from latently infected ganglia, mutant 167 expressed wild-type levels of LAT and the deletion mutant 524 expressed 9- to 15-fold less LAT than normal, while the LAT expression of the LAT- mutant was undetectable or at least 5 log units less than that of the wild type. The rates of recurrence of genital lesions were normal for recombinant viruses 524 and 167 but reduced (as expected) for the LAT- mutant. Alteration of a subset of LAT promoter elements reduced LAT expression by 1 log unit but did not influence the rate of spontaneous disease reactivation in vivo. Far greater reductions in LAT expression are necessary before reactivation rates are noticeably changed.
authors.affiliation: Laboratory of Clinical Investigation, National Institute of Allergy and Infectious Diseases, Bethesda, Maryland 20892, USA. knwang@nih.gov
authors.forename: K
authors.initials: K
authors.lastname: Wang
authors.forename: L
authors.initials: L
authors.lastname: Pesnicak
authors.forename: S E
authors.initials: SE
authors.lastname: Straus
mesh_terms: D000818:Animals; D002522:Chlorocebus aethiops; D017931:DNA Primers; D004279:DNA, Viral / Q000032:analysis; D004848:Epithelium / Q000821:virology; D005260:Female; D005727:Ganglia, Spinal / Q000821:virology; D015967:Gene Expression Regulation, Viral; D006168:Guinea Pigs; D006558:Herpes Genitalis / Q000503:physiopathology*; D018258:Herpesvirus 2, Human / Q000235:genetics / Q000302:isolation & purification / Q000502:physiology*; D016297:Mutagenesis, Site-Directed; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D012194:RNA-Directed DNA Polymerase; D012008:Recurrence; D017384:Sequence Deletion; D012867:Skin / Q000821:virology; D013997:Time Factors; D014158:Transcription, Genetic; D014621:Vagina / Q000821:virology; D014709:Vero Cells; D010948:Viral Plaque Assay; D017735:Virus Latency / Q000235:genetics*; D014779:Virus Replication
chemical_list: D017931:DNA Primers; D004279:DNA, Viral; D012194:RNA-Directed DNA Polymerase