Effect of altering the tRNA(Lys)(3) concentration in human immunodeficiency virus type 1 upon its annealing to viral RNA, GagPol incorporation, and viral infectivity.
Human immunodeficiency virus type 1 (HIV-1) uses tRNA(Lys)(3) as a primer for reverse transcription and, during viral assembly, this tRNA is selectively packaged into the virus along with the other major tRNA(Lys), tRNA(Lys)(3). Increasing the cytoplasmic concentration of tRNA(Lys)(3) through transfection of cells with a plasmid containing both HIV-1 proviral DNA and a tRNA(Lys)(3) gene results in a greater incorporation of tRNA(Lys)(3) into virions, which is accompanied by increased annealing of tRNA(Lys)(3) to the viral genome and increased infectivity of the viral population. Increased viral tRNA(Lys)(3) is accompanied by decreased viral tRNA(Lys)(3), with the total tRNA(Lys)/virion and the GagPol/Gag ratios remaining unchanged. Viral tRNA(Lys) can be doubled, with increases in both tRNA(Lys)(3) and tRNA(Lys)(1,2) concentrations, by overexpressing lysyl tRNA synthetase. This also results in increased tRNA(Lys)(3) annealing to the viral RNA and increased viral infectivity but, again, no change in the GagPol/Gag ratio was observed. This result indicates that GagPol, whose interaction is required during packaging, is not a limiting factor during tRNA(Lys) incorporation into HIV-1, whereas LysRS is.
Lady Davis Institute for Medical Research and McGill AIDS Centre, Jewish General Hospital, McGill University, Montreal, H3T 1E2 Quebec, Canada.
Juliana
J
Gabor
Shan
S
Cen
Hassan
H
Javanbakht
Meijuan
M
Niu
Lawrence
L
Kleiman
D000818:Animals; D019556:COS Cells; D015685:Fusion Proteins, gag-pol / Q000378:metabolism*; D015497:HIV-1 / Q000235:genetics / Q000378:metabolism / Q000472:pathogenicity*; D006367:HeLa Cells; D006801:Humans; D008250:Lysine-tRNA Ligase / Q000235:genetics / Q000378:metabolism; D010957:Plasmids; D012357:RNA, Transfer, Lys / Q000235:genetics / Q000378:metabolism*; D012367:RNA, Viral / Q000378:metabolism*; D014162:Transfection; D019065:Virus Assembly
D015685:Fusion Proteins, gag-pol; D012357:RNA, Transfer, Lys; D012367:RNA, Viral; D008250:Lysine-tRNA Ligase