home
***
CD-ROM
|
disk
|
FTP
|
other
***
search
/
Shareware Overload Trio 2
/
Shareware Overload Trio Volume 2 (Chestnut CD-ROM).ISO
/
dir26
/
med9406b.zip
/
M9460191.TXT
< prev
next >
Wrap
Text File
|
1994-06-12
|
3KB
|
51 lines
Document 0191
DOCN M9460191
TI Transcriptional activation of minimal HIV-1 promoter by ORF-1 protein
expressed from the SalI-L fragment of human herpesvirus 6.
DT 9408
AU Kashanchi F; Thompson J; Sadaie MR; Doniger J; Duvall J; Brady JN;
Rosenthal LJ; Department of Microbiology and Immunology, Georgetown
University; Medical Center, Washington, DC 20007.
SO Virology. 1994 May 15;201(1):95-106. Unique Identifier : AIDSLINE
MED/94233781
AB The SalI-L fragment of human herpesvirus 6 (HHV-6) strain U1102
transformed rodent cells and transactivated the HIV-1 LTR 10- to 15-fold
in both monkey fibroblasts and human T-lymphocytes. In this report, the
SalI-L transactivator of the HIV-1 LTR was localized to ORF-1 which
codes for a protein of 357 amino acids. To determine if ORF-1 required
functional Sp1 binding sites or the TATA box element of HIV-1 LTR for
transactivation, 5'-deletion mutants of the HIV-1 LTR were employed.
Plasmids pBS/SalI-L, pBS/SalI-L-SH, and pC6/ORF-1(S), a mammalian
expression vector containing ORF-1, all transactivated a deletion mutant
of HIV-1 LTR lacking functional Sp1 binding sites (CD-54). These studies
demonstrate that transactivation occurred in the absence of Sp1 binding
sites and required only a minimal HIV-1 promoter which contains the TATA
box element. The specificity of the SalI-L transactivator for HIV-1 LTR
was demonstrated by its inability to transactivate the human
papillomavirus type 16 or 18 early promoters. The ORF-1 gene was cloned
into and expressed from the pET17b bacterial expression vector. Purified
ORF-1 protein was obtained by ammonium sulfate precipitation, Mono-S
chromatography, and anti-T7. Tag immunoaffinity chromatography.
Transactivation of the HIV-1 LTR by ORF-1 protein was demonstrated by
electroporation studies in vivo and by transcription studies in vitro.
To substantiate the putative biological role of ORF-1, pBS/SalI-L,
pBS/SalI-L-SH, and pC6/ORF-1 all reactivated tat-defective HIV-1
provirus from latently infected cells expressing CD4. Thus, the data
presented suggest that HHV-6 infection could have a cofactor role in the
progression of AIDS.
DE Amino Acid Sequence Animal Cell Line Gene Deletion Genes,
Viral/GENETICS Herpesvirus 6, Human/*GENETICS Human HIV Core Protein
p24/ANALYSIS HIV Long Terminal Repeat/*GENETICS
HIV-1/*GENETICS/PHYSIOLOGY Molecular Sequence Data Open Reading
Frames/GENETICS Promoter Regions (Genetics)/GENETICS Recombinant
Fusion Proteins/ANALYSIS/GENETICS Sequence Alignment Support, Non-U.S.
Gov't Trans-Activation (Genetics)/*GENETICS
Trans-Activators/ANALYSIS/CHEMISTRY/*GENETICS Transcription Factor,
Sp1/GENETICS Transfection Tumor Cells, Cultured Viral
Proteins/ANALYSIS/CHEMISTRY/*GENETICS Virus Activation Virus Latency
JOURNAL ARTICLE
SOURCE: National Library of Medicine. NOTICE: This material may be
protected by Copyright Law (Title 17, U.S.Code).