In vivo deletion analysis of the herpes simplex virus type 1 latency-associated transcript promoter.
During herpes simplex virus latency, transcripts accumulate from a single transcription unit of the viral genome. The promoter for these latency-associated transcripts (LAT) has been located, and a number of studies have documented the specific regions of this promoter which are important in transient assays of neuronal cells in culture. To examine the regulation of this promoter from the viral genome, both in vitro and in vivo, a series of seven promoter deletion viruses which drive the expression of the reporter gene beta-galactosidase was constructed. Rabbit skin cells were infected in cell culture with viruses bearing each promoter mutation, and the LAT promoter activity was compared with that obtained by infecting two neuronal cell lines, ND7 cells and C1300 neuroblastoma cells. Mouse dorsal root ganglia were also infected with these recombinant viruses by footpad inoculations, and beta-galactosidase activity was measured. Infected neuronal cells lines and dorsal root ganglia exhibit much more LAT promoter activity than infected rabbit skin cells, suggesting that the region upstream of -250 may contain one or several neuronal specific DNA-binding sites. However, a comparison of LAT promoter activities within the deletion series revealed many differences between neurons of the dorsal root ganglia infected in vivo and the two neuronal cell lines infected in vitro. These results suggest that neurons may vary extensively in the quantity or kind of transcription factors they contain.
Dobson
A T
AT
Department of Microbiology and Immunology, School of Medicine, University of California at Los Angeles 90024.
Margolis
T P
TP
Gomes
W A
WA
Feldman
L T
LT
D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D017931:DNA Primers; D005727:Ganglia, Spinal / Q000378:metabolism / Q000821:virology; D018259:Herpesvirus 1, Human / Q000235:genetics*; D006651:Histocytochemistry; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000235:genetics; D012367:RNA, Viral / Q000235:genetics; D011817:Rabbits; D011995:Recombination, Genetic; D017384:Sequence Deletion; D017735:Virus Latency / Q000235:genetics*
D017931:DNA Primers; D012333:RNA, Messenger; D012367:RNA, Viral