Document 0717
 DOCN  M9650717
 TI    Analysis of trans-dominant mutants of the HIV type 1 Rev protein for
       their ability to inhibit Rev function, HIV type 1 replication, and their
       use as anti-HIV gene therapeutics.
 DT    9605
 AU    Ragheb JA; Bressler P; Daucher M; Chiang L; Chuah MK; Vandendriessche T;
       Morgan RA; National Heart, Lung, and Blood Institute, Bethesda,
       Maryland,; USA.
 SO    AIDS Res Hum Retroviruses. 1995 Nov;11(11):1343-53. Unique Identifier :
       AIDSLINE MED/96159131
 AB    The HIV-1 rev gene product facilitates the transport of singly spliced
       and unspliced HIV-1 transcripts and is necessary for productive HIV-1
       infection. On the basis of the previously described trans-dominant Rev
       mutant M10, four point mutants and one frameshift mutant of the Rev
       protein were constructed. The mutants were inserted into retroviral
       expression vectors and analyzed for their ability to inhibit
       Rev-mediated gene expression. Transient transfection systems were used
       to screen these new mutants, and each was shown to inhibit expression of
       a Rev-dependent CAT reporter plasmid. Inhibition of HIV-1 envelope gene
       expression was tested in the HeLa-T4 cell line and was also shown to be
       inhibited by the trans-dominant Rev mutants. Retroviral vector producer
       cell lines were constructed and used to transduce Rev trans-dominant
       genes into the human T-cell line SupT1. The engineered SupT1 cell lines
       were then challenged with HIV-1 IIIB and HIV-1 expression was monitored
       by Northern blot analysis and in situ hybridization. SupT1 cells
       expressing either a Rev point mutant or the frameshift mutant showed
       greatly reduced HIV-1 mRNA accumulation and the Rev-dependent singly
       spliced and unspliced HIV-1 mRNAs were reduced. The kinetics of viral
       replication following challenge of Rev trans-dominant-engineered SupT1
       cells with both HIV-1 IIIB and MN strains was significantly reduced and
       cells were protected from viral lysis. Viruses that emerge late in
       infection from Rev trans-dominant-engineered cultures are not resistant
       to Rev-mediated inhibition. Last, trans-dominant Rev-mediated protection
       of human CD4+ lymphocytes from challenge with primary HIV-1 patient
       isolates confirms the potential utility of this system as an anti-HIV-1
       gene therapy approach.
 DE    Amino Acid Sequence  Animal  Base Sequence  Cell Line  Chloramphenicol
       Acetyltransferase/GENETICS  Frameshift Mutation  Gene Products,
       env/GENETICS  Gene Products, rev/*GENETICS/THERAPEUTIC USE  *Gene
       Therapy  Genes, Dominant  Genes, Reporter  Hela Cells  Human  HIV
       Infections/*THERAPY/VIROLOGY  HIV-1/*GENETICS/PHYSIOLOGY  Molecular
       Sequence Data  *Mutation  Point Mutation  RNA, Messenger/METABOLISM
       RNA, Viral/METABOLISM  Support, Non-U.S. Gov't  Transfection  Virus
       Replication/PHYSIOLOGY  JOURNAL ARTICLE

       SOURCE: National Library of Medicine.  NOTICE: This material may be
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