       Document 0625
 DOCN  M9630625
 TI    Analysis of trafficking of Rev and transdominant Rev proteins in living
       cells using green fluorescent protein fusions: transdominant Rev blocks
       the export of Rev from the nucleus to the cytoplasm.
 DT    9603
 AU    Stauber R; Gaitanaris GA; Pavlakis GN; Human Retrovirus Section,
       NCI-FCRDC, Maryland 21702-1201, USA.
 SO    Virology. 1995 Nov 10;213(2):439-49. Unique Identifier : AIDSLINE
       MED/96074519
 AB    Expression of gag/pol and env genes of human immunodeficiency virus
       requires the viral Rev protein. Mutant Rev proteins, displaying a
       transdominant phenotype (TDRev), were shown to inhibit Rev function. To
       investigate the underlying mechanism of this inhibition, the green
       fluorescent protein (GFP) of Aequorea victoria was fused to Rev and
       TDRev, which allowed the study of their trafficking and interactions in
       living human cells. Both Rev-GFP and TDRev-GFP were shown to retain
       appropriate nucleolar localization and function. Upon actinomycin D
       treatment, Rev-GFP was transported to the cytoplasm within 1.5 hr, while
       TDRev, although partially dissociated from the nucleolus, was retained
       in the nucleus. Coexpression of Rev-GFP and TDRev in the same cell
       demonstrated that TDRev inhibited the transport of Rev-GFP from the
       nucleus to the cytoplasm. This inhibition was specific for Rev, since
       the export of the functionally analogous Rex protein of the human T-cell
       leukemia virus type I was not inhibited by TDRev. These results indicate
       that Rev and TDRev form heteromultimers in the nucleolus and that this
       interaction prevents Rev's export from the nucleus to the cytoplasm. In
       addition to providing a model for the function of TDRev, these results
       also demonstrate the successful application of protein fusions to GFP to
       study localization and trafficking of proteins in living mammalian
       cells.
 DE    Base Sequence  Biological Transport  Cell Line, Transformed  Cell
       Nucleolus/METABOLISM  Cell Nucleus/*METABOLISM  Cytoplasm/*METABOLISM
       Dactinomycin/PHARMACOLOGY  DNA Primers  Gene Products,
       rev/GENETICS/*METABOLISM  Gene Products, rex/METABOLISM  Hela Cells
       Human  HIV Core Protein p24/BIOSYNTHESIS  HIV-1/*METABOLISM  Luminescent
       Proteins/GENETICS/METABOLISM  Molecular Sequence Data  Phenotype
       Recombinant Fusion Proteins/METABOLISM  Support, Non-U.S. Gov't
       Support, U.S. Gov't, P.H.S.  JOURNAL ARTICLE

       SOURCE: National Library of Medicine.  NOTICE: This material may be
       protected by Copyright Law (Title 17, U.S.Code).

