Analysis of a herpes simplex virus type 1 LAT mutant with a deletion between the putative promoter and the 5' end of the 2.0-kilobase transcript.
A herpes simplex virus type 1 strain 17 mutant with a deletion between genomic nucleotides 118880 and 119250 was constructed and called 17 delta Sty. The deletion removes most of a putative secondary LAT promoter (called LAPII) as well as 370 of the first 449 nucleotides of the proposed 8.5-kb transcript believed to be the precursor of 2.0-kb LAT. 17 delta Sty was shown to produce major 2.0-kb LATs in tissue culture. Moreover, trigeminal nerves from latently infected mice contained an intact 1.45- to 2.0-kb LAT as well as the minor LATs which are recognized by probes specific for regions downstream of the 2.0-kb LAT. Finally, 17 delta Sty reactivated with normal kinetics from the trigeminal ganglia of latently infected mice in the explant cocultivation assay and egressed from tissue culture cells as efficiently as wild-type virus. These results clearly show that the region deleted in 17 delta Sty is dispensable for intact 2-kb LAT production, viral egress in tissue culture, and normal reactivation from latently infected neurons in mice.
Department of Microbiology and Immunology, Jefferson Cancer Institute, Thomas Jefferson University, Philadelphia, Pennsylvania 19107-6799.
J
J
Maggioncalda
A
A
Mehta
N W
NW
Fraser
T M
TM
Block
D000818:Animals; D015152:Blotting, Northern; D002460:Cell Line; D002522:Chlorocebus aethiops; D004279:DNA, Viral / Q000302:isolation & purification; D006561:Herpes Simplex / Q000473:pathology / Q000503:physiopathology; D018259:Herpesvirus 1, Human / Q000235:genetics* / Q000254:growth & development; D007668:Kidney; D051379:Mice; D008807:Mice, Inbred BALB C; D009474:Neurons / Q000473:pathology / Q000821:virology; D009924:Organ Culture Techniques; D011401:Promoter Regions, Genetic; D012367:RNA, Viral / Q000302:isolation & purification; D015183:Restriction Mapping; D017384:Sequence Deletion; D013997:Time Factors; D014158:Transcription, Genetic; D012668:Trigeminal Ganglion / Q000473:pathology / Q000821:virology; D014774:Virulence; D014775:Virus Activation
D004279:DNA, Viral; D012367:RNA, Viral