Expression of active human immunodeficiency virus type 1 protease by noninfectious chimeric virus particles.
To generate nonpathogenic viral particles which express active human immunodeficiency virus type 1 (HIV-1) protease (PR), plasmids containing sequences from the genomes of HIV-1 and Moloney murine leukemia virus (M-MuLV) were constructed. Either the PR coding region alone; the gag, PR, and reverse transcriptase protein-coding regions; or the complete gag and pol protein-coding regions from HIV-1 were substituted for the corresponding regions of a full-length M-MuLV clone to yield the chimeric plasmids pMoHIV-I, pMoHIV-III, and pMoHIV-IV, respectively. Cell lines which express the viral gag polyprotein were isolated for hybrids pMoHIV-I and pMoHIV-III. These cells produced viral particles which contained processed core proteins. Cleavage of the gag polyprotein in the viral particles was inhibited by the HIV-1 PR inhibitor L-687908, indicating that the viral PR is responsible for the observed processing. The hybrid virions were not infectious; analyses indicated that the viral particles contained little or no reverse transcriptase activity. In addition, particles produced by pMoHIV-III transfectants failed to package the viral genomic RNA. The cell line which expresses and processes the HIV-1 gag polyprotein is a safe and effective reagent for the in vivo evaluation of potential inhibitors of the HIV-1 PR.
Kohl
N E
NE
Department of Molecular Biology, Merck Sharp and Dohme Research Laboratories, West Point, Pennsylvania 19486.
Diehl
R E
RE
Rands
E
E
Davis
L J
LJ
Hanobik
M G
MG
Wolanski
B
B
Dixon
R A
RA
D000818:Animals; D002460:Cell Line; D002678:Chimera; D004279:DNA, Viral / Q000737:chemistry; D004353:Drug Evaluation, Preclinical / Q000379:methods; D015870:Gene Expression; D016333:HIV Protease / Q000096:biosynthesis / Q000235:genetics*; D015497:HIV-1 / Q000201:enzymology / Q000235:genetics*; D006868:Hydrolysis; D051379:Mice; D008815:Mice, Inbred Strains; D008979:Moloney murine leukemia virus / Q000235:genetics / Q000648:ultrastructure; D010957:Plasmids; D012367:RNA, Viral / Q000378:metabolism; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism; D014162:Transfection; D014771:Virion / Q000235:genetics / Q000648:ultrastructure
D004279:DNA, Viral; D012367:RNA, Viral; D012194:RNA-Directed DNA Polymerase; D016333:HIV Protease