PMID- 14581568
OWN - NLM
STAT- MEDLINE
DCOM- 20031125
LR  - 20190509
IS  - 0022-538X (Print)
IS  - 1098-5514 (Electronic)
IS  - 0022-538X (Linking)
VI  - 77
IP  - 22
DP  - 2003 Nov
TI  - Analysis of herpes simplex virus ICP0 promoter function in sensory neurons during 
      acute infection, establishment of latency, and reactivation in vivo.
PG  - 12319-30
AB  - We have begun an analysis of the functional architecture of the ICP0 promoter in 
      neurons in vivo with the ultimate goal of determining how this gene is regulated 
      during reactivation in vivo. Promoter/reporter mutants in which the Escherichia 
      coli beta-galactosidase (beta-Gal) gene was driven by various permutations of the 
      ICP0 promoter were employed to permit the analysis of promoter function without 
      the added complications that would arise due to inappropriate regulation of ICP0 
      protein levels. A whole-ganglion immunohistochemical staining procedure (N. M. 
      Sawtell, J. Virol. 77:4127-4138, 2003) was used for direct comparisons of the 
      expression of the promoter/reporter gene to expression of the native protein in 
      the same cell. In this way, the expression of the putative wild-type promoter 
      could be validated and results for mutant promoters could be compared to 
      expression of the native gene. We found that a DNA fragment from bp -562 through 
      the methionine start codon of the ICP0 gene contained all sequences required for 
      properly regulated ICP0 expression in diverse cell types (including sensory 
      neurons of the trigeminal ganglia [TG]) in vitro and in vivo, as indicated by 
      colocalization of ICP0 and beta-Gal. Truncation of the ICP0 promoter to bp -145 
      or -129 resulted in the loss of immediate-early (alpha) kinetics. The truncated 
      promoters expressed high levels of the reporter gene with leaky late (gamma1) 
      kinetics in vitro and in some cell types in vivo. Unexpectedly, the truncated 
      promoters did not express in TG neurons. Thus, TAATGARAT or other sequences 
      upstream of bp -145 in the ICP0 promoter are required for basal expression of 
      ICP0 in neurons but are not required for basal expression in other cells in vivo. 
      There was a >95% concordance between reporter and native protein expression 
      detected with the 562-bp promoter in neurons during the acute stage. However, 
      this was not the case during reactivation from latency in vivo, as nearly twice 
      as many neurons contained detectable beta-Gal as contained detectable ICP0. This 
      same 562-bp promoter/reporter cassette, when placed in the context of a 
      latency-associated transcript (LAT) null mutant, resulted in >95% concordance of 
      expression of beta-Gal and ICP0 during reactivation in vivo. These last results 
      strongly suggest that there is a posttranscriptional constraint on the expression 
      of ICP0 protein during reactivation from latency and that this constraint is 
      mediated by LAT.
FAU - Thompson, R L
AU  - Thompson RL
AD  - Department of Molecular Genetics, Biochemistry, and Microbiology, University of 
      Cincinnati Medical Center, Cincinnati, Ohio 45267-0524, USA. 
      Richard.Thompson@UC.edu
FAU - Shieh, May T
AU  - Shieh MT
FAU - Sawtell, N M
AU  - Sawtell NM
LA  - eng
GR  - R01 AI032121/AI/NIAID NIH HHS/United States
GR  - R01 EY013168/EY/NEI NIH HHS/United States
GR  - R01 AI 32121/AI/NIAID NIH HHS/United States
GR  - R01 EY 13168/EY/NEI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - J Virol
JT  - Journal of virology
JID - 0113724
RN  - 0 (Immediate-Early Proteins)
RN  - EC 2.3.2.27 (Ubiquitin-Protein Ligases)
RN  - EC 2.3.2.27 (Vmw110 protein, Human herpesvirus 1)
RN  - EC 3.2.1.23 (beta-Galactosidase)
SB  - IM
MH  - DNA Replication
MH  - Herpesvirus 1, Human/genetics/*physiology
MH  - Immediate-Early Proteins/*genetics
MH  - Promoter Regions, Genetic/*physiology
MH  - Transcription, Genetic
MH  - Trigeminal Ganglion/*virology
MH  - Ubiquitin-Protein Ligases
MH  - *Virus Activation
MH  - *Virus Latency
MH  - *Virus Replication
MH  - beta-Galactosidase/analysis
PMC - PMC254249
EDAT- 2003/10/29 05:00
MHDA- 2003/12/03 05:00
PMCR- 2003/11/01
CRDT- 2003/10/29 05:00
PHST- 2003/10/29 05:00 [pubmed]
PHST- 2003/12/03 05:00 [medline]
PHST- 2003/10/29 05:00 [entrez]
PHST- 2003/11/01 00:00 [pmc-release]
AID - 0549 [pii]
AID - 10.1128/jvi.77.22.12319-12330.2003 [doi]
PST - ppublish
SO  - J Virol. 2003 Nov;77(22):12319-30. doi: 10.1128/jvi.77.22.12319-12330.2003.