Document 0203
 DOCN  M9460203
 TI    Assembly of HIV GAG-B-galactosidase fusion proteins into virus
       particles.
 DT    9408
 AU    Wang CT; Stegeman-Olsen J; Zhang Y; Barklis E; Vollum Institute for
       Advanced Biomedical Research, Oregon Health; Sciences University,
       Portland 97201.
 SO    Virology. 1994 May 1;200(2):524-34. Unique Identifier : AIDSLINE
       MED/94233715
 AB    We have studied the assembly of human immunodeficiency virus (HIV-1)
       Gag-B-galactosidase (Gag-B-gal; GBG) fusion proteins into HIV particles
       in the presence of HIV Gag proteins. Release of fusion proteins from
       cells was measured by assay of media versus cellular B-gal activities
       and was dependent on co-expression of unfused Gag proteins. Gag-B-gal
       incorporation into virus particles was demonstrated by detergent
       treatment and density gradient fractionation studies and was dependent
       on protein-protein interactions requiring the C-terminal two-thirds of
       the HIV CA domain. The central MA domain appeared unimportant for fusion
       protein incorporation; a nonmyristylated GBG protein was incorporated
       but at a relatively reduced level, while the NC and p6 domains slightly
       affected the assembly of fusion proteins into particles. Subcellular
       fractionation studies showed that all fusion proteins including the
       nonmyristylated one were enriched in the cytoplasmic pellet fraction.
       However, assembly into particles did not correlate with subcellular
       fractionation patterns. Similarly, virion incorporation levels of
       Gag-B-gal proteins did not correlate with their immunofluorescence
       localization patterns. However, we observed that while most fusion
       proteins displayed a perinuclear ring with heterogeneous staining
       throughout cells, short fusion proteins appeared enriched on the
       intracellular membranes, and fusion proteins with intact MA but deleted
       NC domains showed an enhanced surface staining without a clear
       perinuclear ring. Altogether, our data suggest that the CA domain is the
       primary determinant for assembly of HIV fusion proteins into virus
       particles.
 DE    beta-Galactosidase/ANALYSIS/METABOLISM  Animal  Base Sequence  Cell
       Compartmentation  Comparative Study  Gene Products,
       gag/GENETICS/*METABOLISM  HIV-1/*GROWTH & DEVELOPMENT  Molecular
       Sequence Data  Morphogenesis/GENETICS  Myristic Acids/METABOLISM
       Protein Processing, Post-Translational  Recombinant Fusion
       Proteins/METABOLISM  Structure-Activity Relationship  Support, Non-U.S.
       Gov't  Support, U.S. Gov't, P.H.S.  Transfection  Virion/*GROWTH &
       DEVELOPMENT  JOURNAL ARTICLE

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