title: Neither LAT nor open reading frame P mutations increase expression of spliced or intron-containing ICP0 transcripts in mouse ganglia latently infected with herpes simplex virus.
abstract: Latent infections by herpes simplex virus are characterized by repression of productive-cycle gene expression. Several hypotheses to explain this repression involve inhibition of expression of the immediate-early gene activator ICP0 during latency. To address these hypotheses, we developed quantitative reverse transcriptase-PCR assays that detected spliced and intron-containing ICP0 transcripts in mouse ganglia latently infected with wild-type virus. In these ganglia, the numbers of spliced ICP0 transcripts correlated better with the numbers of transcripts from the immediate-early gene encoding ICP4 than with those from the early gene encoding thymidine kinase. There were fewer spliced than intron-containing ICP0 transcripts on average, with considerable ganglion-to-ganglion variation. We then investigated whether ICP0 expression in latently infected ganglia is reduced by the latency-associated transcripts (LATs) and whether splicing of ICP0 transcripts is inhibited by the product of open reading frame (ORF) P. A LAT deletion mutation which essentially eliminates expression of the major LATs did not appreciably increase levels of ICP0 transcripts. LAT deletion mutants did, however, appear to express reduced levels of intron-containing ICP0 transcripts. ORF P mutations did not alter levels of ICP0 transcripts in a manner consistent with inhibition of ICP0 splicing by ORF P. Although these results argue against antisense inhibition of ICP0 expression by LATs or inhibition of ICP0 splicing by ORF P, they are consistent with the possibilities of a block between immediate-early and early gene expression and regulation of spliced versus intron-containing ICP0 transcripts in latently infected ganglia.
authors.affiliation: Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 02115, USA. shunhua@mail.ncku.edu.tw
authors.forename: Shun-Hua
authors.initials: SH
authors.lastname: Chen
authors.forename: Lily Yeh
authors.initials: LY
authors.lastname: Lee
authors.forename: David A
authors.initials: DA
authors.lastname: Garber
authors.forename: Priscilla A
authors.initials: PA
authors.lastname: Schaffer
authors.forename: David M
authors.initials: DM
authors.lastname: Knipe
authors.forename: Donald M
authors.initials: DM
authors.lastname: Coen
mesh_terms: D000818:Animals; D004279:DNA, Viral / Q000032:analysis; D004195:Disease Models, Animal; D015967:Gene Expression Regulation, Viral; D006561:Herpes Simplex / Q000821:virology*; D018259:Herpesvirus 1, Human / Q000235:genetics / Q000502:physiology*; D017874:Immediate-Early Proteins / Q000032:analysis / Q000235:genetics*; D007438:Introns; D008297:Male; D051379:Mice; D009154:Mutation; D016366:Open Reading Frames; D012333:RNA, Messenger / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013937:Thymidine Kinase / Q000032:analysis / Q000235:genetics; D012668:Trigeminal Ganglion / Q000821:virology*; D044767:Ubiquitin-Protein Ligases; D014764:Viral Proteins / Q000032:analysis / Q000235:genetics*; D014775:Virus Activation; D017735:Virus Latency / Q000235:genetics
chemical_list: D004279:DNA, Viral; D017874:Immediate-Early Proteins; C101627:ORF P protein, herpes simplex virus; D012333:RNA, Messenger; D014764:Viral Proteins; C048925:herpes simplex virus, type 1 protein ICP4; D044767:Ubiquitin-Protein Ligases; C051804:Vmw110 protein, Human herpesvirus 1; D013937:Thymidine Kinase