title: Herpes simplex virus 2 microRNA miR-H6 is a novel latency-associated transcript-associated microRNA, but reduction of its expression does not influence the establishment of viral latency or the recurrence phenotype.
abstract: The herpes simplex virus 2 (HSV-2) viral microRNA (miRNA) designated miR-H6 is located upstream of the latency-associated transcript (LAT) promoter region on the strand opposite the LAT. Deletion of the LAT promoter and part of LAT exon 1 abolished HSV-2 miR-H6 expression in acutely and latently infected guinea pig dorsal root ganglia (DRG), suggesting that this region is needed both for the expression of LAT-encoded miRNAs and for miR-H6 expression in vivo. Relative to cells infected with a viral rescuant, miR-H6 expression was significantly reduced in cells infected with a mutant HSV-2 virus, NotPolyA, with an insertion of a simian virus (SV40) polyadenylation signal sequence between the LAT promoter and miR-H6 sequences. In addition, expression of miR-H6, but not LAT or viral DNA, was significantly reduced in both mouse trigeminal ganglia (TG) and guinea pig DRG latently infected with the NotPolyA mutant. Guinea pigs infected with NotPolyA experienced reduced neurological complications of acute infection relative to those infected with the rescuant, but the recurrence phenotype of the NotPolyA mutant was similar to those of its rescuant and wild-type HSV-2, indicating that reduction of miR-H6 expression is not by itself able to alter the establishment of latency for the wild-type virus or the recurrence phenotype. Furthermore, the mutation in NotPolyA did not affect the propensity of wild-type HSV-2 to establish latency in neurons positive for subtype marker KH10. In contrast to published reports regarding its HSV-1 homolog, HSV-2 miR-H6 did not affect ICP4 expression in transfected or infected cells. We hypothesize that viral miRNAs associated with LAT expression are likely to work collectively, contributing to the phenotype attributed to the LAT.
authors.affiliation: Division of Viral Products, Office of Vaccines Research and Review, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, Maryland 20892, USA.
authors.forename: Shuang
authors.initials: S
authors.lastname: Tang
authors.forename: Andrea S
authors.initials: AS
authors.lastname: Bertke
authors.forename: Amita
authors.initials: A
authors.lastname: Patel
authors.forename: Todd P
authors.initials: TP
authors.lastname: Margolis
authors.forename: Philip R
authors.initials: PR
authors.lastname: Krause
mesh_terms: D000818:Animals; D005260:Female; D005727:Ganglia, Spinal / Q000821:virology; D015967:Gene Expression Regulation, Viral; D006168:Guinea Pigs; D018258:Herpesvirus 2, Human / Q000235:genetics / Q000502:physiology*; D051379:Mice; D035683:MicroRNAs / Q000378:metabolism*; D012367:RNA, Viral / Q000378:metabolism*; D014764:Viral Proteins / Q000096:biosynthesis / Q000235:genetics; D017735:Virus Latency
chemical_list: C477254:LAT protein, Human herpesvirus 2; D035683:MicroRNAs; D012367:RNA, Viral; D014764:Viral Proteins