Herpes simplex virus type 1 deletion variants 1714 and 1716 pinpoint neurovirulence-related sequences in Glasgow strain 17+ between immediate early gene 1 and the 'a' sequence.
Dideoxynucleotide sequence analysis of a spontaneously isolated deletion variant (1714) of Glasgow strain 17+ of herpes simplex virus type 1 (HSV-1) demonstrates that the deletion is 759 bp in length and is located within each copy of the BamHI s fragment (0 to 0.02 and 0.81 to 0.83 map units) of the long repeat region of the genome. The deletion removes one complete copy of the 18 bp DR1 element of the 'a' sequence and terminates 1105 bp upstream of the 5' end of immediate early (IE) gene 1. The variant grows to high titre, is not temperature-sensitive and is not host cell type-restricted in vitro. In vivo studies demonstrate that 1714 is totally avirulent for BALB/c mice following intracerebral inoculation, with an LD50 of 7 x 10(6) p.f.u./mouse compared to less than 10 p.f.u./mouse for the parental wild-type strain 17+. In vivo growth kinetics show that the non-neurovirulent phenotype is due to an inability to replicate in mouse brain. Because 1714 was in a genomic background in which the four XbaI sites had been removed and because the phenotype was thymidine kinase-negative, the 759 bp deletion was introduced into an otherwise totally wild-type background. The resulting variant (1716) is non-neurovirulent for mice, with an LD50 of 7 x 10(6) p.f.u./mouse. The deletion does not prevent the virus from establishing a latent infection or reactivating from it in vitro. The results demonstrate that sequences between IE-1 and the 'a' sequence produce neurovirulence in Glasgow strain 17+ and, in conjunction with the non-neurovirulence of the HSV-2 HG52 variant JH2604, identify a common function conserved in HSV-1 and -2.
MacLean
A R
AR
MRC Virology Unit, Institute of Virology, Glasgow, U.K.
ul-Fareed
M
M
Robertson
L
L
Harland
J
J
Brown
S M
SM
D000818:Animals; D001345:Autoradiography; D001483:Base Sequence; D015139:Blotting, Southern; D001921:Brain / Q000382:microbiology*; D002460:Cell Line; D004279:DNA, Viral / Q000737:chemistry; D015245:Deoxyribonuclease BamHI; D004660:Encephalitis / Q000382:microbiology*; D006561:Herpes Simplex / Q000382:microbiology*; D006801:Humans; D007700:Kinetics; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D015183:Restriction Mapping; D018139:Simplexvirus / Q000235:genetics* / Q000254:growth & development / Q000472:pathogenicity; D014709:Vero Cells; D014774:Virulence / Q000235:genetics
D004279:DNA, Viral; D015245:Deoxyribonuclease BamHI