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).