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M94A0164.TXT
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1994-10-01
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Document 0164
DOCN M94A0164
TI Cell line-dependent release of HIV-like gag particles after infection of
mammalian cells with recombinant vaccinia viruses.
DT 9412
AU Dru A; Ludosky MA; Cartaud J; Beaud G; Institut Jacques Monod, Paris,
France.
SO AIDS Res Hum Retroviruses. 1994 Apr;10(4):383-90. Unique Identifier :
AIDSLINE MED/94347462
AB We investigated the production of Gag particles by Vero, CV-1, or 1D
cells infected with different vaccinia virus recombinants expressing HIV
gag or gag-pol genes. Immunoblots of (centrifuged) culture media from 1D
cells infected with vMM5, a vaccinia virus recombinant expressing the
HIV-2 gag-pol genes, revealed the presence of abundant particles that
contained (mostly processed) Gag antigens. In contrast, Gag particles
were found only in low amounts in the culture medium from Vero cells
infected with the same HIV gag-pol vaccinia virus recombinant; the Gag
precursor remained associated with the infected Vero cells and was
efficiently processed. This low excretion of Gag particles after
infection of Vero cells with vMM5 was also demonstrated by assays of
reverse transcriptase activity in the pellet of centrifuged culture
medium. Cell fractionation showed that Gag proteins were predominantly
found in the membrane fraction from both 1D and Vero cells. Electron
microscopy observations of 1D or of Vero cells infected with vMM5
vaccinia virus recombinant revealed in both cases the presence of
particles budding at the plasma membrane. However, the shape of the
budding particles was different in the two cell lines, with immature
forms present in the membrane from the infected Vero cells. An
inefficient excretion of Gag particles was also observed after infection
of Vero cells with different vaccinia virus recombinants expressing
either an uncleaved HIV-2 Gag protein or the HIV-1 gag-pol genes, as
judged both by immunoblot and reverse transcriptase activity
assays.(ABSTRACT TRUNCATED AT 250 WORDS)
DE Animal Cell Line Gene Expression Gene Products,
gag/*BIOSYNTHESIS/GENETICS Genes, gag Genes, pol HIV-1/*GENETICS
HIV-2/*GENETICS Mice Microscopy, Electron Myristic Acids/METABOLISM
Protein Precursors/BIOSYNTHESIS/GENETICS Protein Processing,
Post-Translational Recombination, Genetic Reverse
Transcriptase/METABOLISM Support, Non-U.S. Gov't Vaccinia
Virus/*GENETICS Vero Cells JOURNAL ARTICLE
SOURCE: National Library of Medicine. NOTICE: This material may be
protected by Copyright Law (Title 17, U.S.Code).