Expression of chimeric neo-Rev response element sequences interferes with Rev-dependent HIV-1 Gag expression.
Recombinant plasmids containing reiterated human immunodeficiency virus type 1 (HIV-1) Rev response element (RRE) sequences were constructed to suppress Rev-dependent HIV-1 Gag expression. The mammalian expression vectors pMAMneo containing one, three, or six repeats of the RRE sequence were cotransfected with a HIV-1 HTLV-IIIB proviral DNA into HeLa cells. All three RRE expression plasmids reduced replication of HIV-1 with similar efficacy. Furthermore, the chimeric expression vector pCMV neoRRE6 x ----(containing six copies of the RRE sequence) was used to establish HeLa cell lines constitutively expressing RRE. A plasmid encoding a Rev-dependent HIV-1 p24 Gag protein was cotransfected with the wild-type Rev expression plasmid into three different RRE-expressing HeLa cell lines. p24 Gag protein production in the culture supernatants of the HeLaneoRRE cells was compared with two neo-expressing cell lines. Although all cell lines (HeLaneoRRE, HeLaneo) displayed similar transfection efficiencies, p24 Gag protein synthesis was markedly reduced in the RRE-expressing cell lines in comparison to the control cells.
Zimmermann
K
K
SANDOZ Research Institute, Vienna, Austria.
Weber
S
S
Dobrovnik
M
M
Hauber
J
J
Böhnlein
E
E
D001483:Base Sequence; D002678:Chimera / Q000235:genetics; D004279:DNA, Viral; D015967:Gene Expression Regulation, Viral; D015752:Genes, env; D015757:Genes, rev; D016655:HIV Core Protein p24 / Q000096:biosynthesis / Q000235:genetics*; D015497:HIV-1 / Q000235:genetics*; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D010957:Plasmids; D012367:RNA, Viral / Q000096:biosynthesis
D004279:DNA, Viral; D016655:HIV Core Protein p24; D012367:RNA, Viral