Detection of latency-related viral RNAs in trigeminal ganglia of rabbits latently infected with herpes simplex virus type 1.
Using a combination of in situ hybridization and Northern (RNA) blot analysis, we investigated herpes simplex virus type 1 (HSV-1) transcriptional activity in an ocular rabbit model of HSV-1 latency. Radioactively labeled cloned fragments, representing virtually the entire HSV-1 genome, were individually hybridized to RNA in sections of trigeminal ganglia taken from rabbits during the latent phase of infection with HSV-1 (McKrae). Our results suggest that two discrete latency-related RNAs (LR-RNAs) may be present. The LR-RNAs were localized mainly in the nuclei of neurons. The more abundant LR-RNA was detected in approximately 3% of all neurons examined and was designated major LR-RNA. The other LR-RNA, designated minor LR-RNA, was detected in approximately 0.3% of neurons from latently infected rabbits. The genes for the LR-RNAs mapped in the vicinity of the immediate-early gene ICP0 (also designated IE110). The gene for the major LR-RNA partially overlapped the left (3') end of the ICP0 gene. In situ hybridization with single-stranded RNA probes showed that this LR-RNA was of complementary sense to that of ICP0 mRNA. Northern blot analysis gave an approximate size for this LR-RNA of 1.8 to 2.2 kilobases. The minor LR-RNA mapped to or near the right (5') end of the ICP0 gene. The detection of LR-RNAs suggests the possibility that these RNAs or their products may play significant roles in the initiation and/or maintenance of HSV-1 latency.
Rock
D L
DL
Department of Veterinary Science, North Dakota State University, Fargo 50105.
Nesburn
A B
AB
Ghiasi
H
H
Ong
J
J
Lewis
T L
TL
Lokensgard
J R
JR
Wechsler
S L
SL
D000818:Animals; D003001:Cloning, Molecular; D004262:DNA Restriction Enzymes; D004279:DNA, Viral / Q000235:genetics; D015245:Deoxyribonuclease BamHI; D015246:Deoxyribonuclease EcoRI; D004195:Disease Models, Animal; D005814:Genes, Viral; D006561:Herpes Simplex / Q000382:microbiology*; D008297:Male; D009693:Nucleic Acid Hybridization; D012367:RNA, Viral / Q000032:analysis* / Q000096:biosynthesis / Q000235:genetics; D011817:Rabbits; D012091:Repetitive Sequences, Nucleic Acid; D018139:Simplexvirus / Q000235:genetics*; D014158:Transcription, Genetic; D012668:Trigeminal Ganglion / Q000382:microbiology*; D014276:Trigeminal Nerve / Q000382:microbiology*
D004279:DNA, Viral; D012367:RNA, Viral; D004262:DNA Restriction Enzymes; D015245:Deoxyribonuclease BamHI; D015246:Deoxyribonuclease EcoRI