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M9490416.TXT
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1994-09-19
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Document 0416
DOCN M9490416
TI The herpes simplex virus type 1 major capsid protein (VP5-UL19) promoter
contains two cis-acting elements influencing late expression.
DT 9411
AU Huang CJ; Wagner EK; Department of Molecular Biology and Biochemistry,
University of; California, Irvine 92717.
SO J Virol. 1994 Sep;68(9):5738-47. Unique Identifier : AIDSLINE
MED/94335089
AB The herpes simplex virus type 1 (HSV-1) major capsid protein VP5 gene
(UL19) is expressed with beta gamma (gamma 1 [leaky late]) kinetics. We
have previously described the construction of recombinant HSV-1 in which
the VP5 promoter was engineered to control the expression of the
bacterial beta-galactosidase gene as a reporter (C.-J. Huang, S. A.
Goodart, M. K. Rice, J. F. Guzowski, and E. K. Wagner, J. Virol.
67:5109-5116, 1993). Here we describe further mutational analysis in
recombinant viruses. We have precisely defined the boundaries of the VP5
promoter and identified two regions important for both the level and the
kinetics of expression. The 5' boundary was located at -48 relative to
the initiation site of transcription by analyzing a series of nested
deletions in the upstream sequence, and although a number of cis-acting
sites influencing transient expression have been identified upstream of
this point, these sites have no role in promoter activity during
productive infection. Deletion of an Sp1-binding site located between
-48 and the TATA box at 30 greatly reduced VP5 promoter activity late
but not early after infection. A cis-acting element whose sequence
resembles the human immunodeficiency virus type 1 initiator was located
between -2 and +10 in the VP5 sequence by characterizing a series of
deletions and site-directed block mutations downstream the TATA box.
This element defines the 3' limit of the VP5 promoter, and like the
upstream element, disruption of this element also inhibited promoter
activity late in the productive cycle.
DE Base Sequence Binding Sites Capsid/*GENETICS Consensus Sequence DNA
Mutational Analysis *Gene Expression Regulation, Viral Herpesvirus 1,
Human/*GENETICS Molecular Sequence Data Mutagenesis, Site-Directed
Promoter Regions (Genetics) RNA, Messenger/GENETICS Sequence Deletion
Support, U.S. Gov't, P.H.S. Transcription Factor, Sp1/METABOLISM
Transcription, Genetic JOURNAL ARTICLE
SOURCE: National Library of Medicine. NOTICE: This material may be
protected by Copyright Law (Title 17, U.S.Code).