Ability of wild-type and mutant lysyl-tRNA synthetase to facilitate tRNA(Lys) incorporation into human immunodeficiency virus type 1.
The major human tRNA(Lys) isoacceptors, tRNA1,2Lys and tRNA3Lys, are selectively packaged into human immunodeficiency virus type 1 (HIV-1) during assembly, where tRNA3Lys acts as a primer for reverse transcription. Lysyl-tRNA synthetase (LysRS) is also incorporated into HIV-1, independently of tRNA(Lys), via its interaction with Gag, and it is a strong candidate for being the signal that specifically targets tRNA(Lys) for viral incorporation. Expression of exogenous wild-type LysRS in cells results in an approximately twofold increase in the viral packaging of both LysRS and tRNA(Lys). Herein, we show that this increase in tRNA(Lys) incorporation into virions is dependent upon the ability of LysRS to bind to tRNA(Lys) but not upon its ability to aminoacylate the tRNA(Lys). COS7 cells were cotransfected with plasmids coding for both HIV-1 and either wild-type or mutant human LysRS, all of which are incorporated into virions with similar efficiency. However, N-terminally truncated LysRS, which binds poorly to tRNA(Lys), does not increase tRNA(Lys) packaging into viruses, while C-terminally truncated LysRS, which binds to but does not aminoacylate tRNA(Lys), still facilitates an increase in tRNA(Lys) packaging into virions.
Cen
Shan
S
Lady Davis Institute for Medical Research and McGill AIDS Centre, Jewish General Hospital. Departments of Medicine. Microbiology and Immunology, McGill University, Montreal, Quebec, Canada H3T 1E2.
Javanbakht
Hassan
H
Niu
Meijuan
M
Kleiman
Lawrence
L
D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D003593:Cytoplasm / Q000201:enzymology; D015497:HIV-1 / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008250:Lysine-tRNA Ligase / Q000235:genetics / Q000378:metabolism*; D009154:Mutation; D010957:Plasmids; D012357:RNA, Transfer, Lys / Q000378:metabolism*; D014162:Transfection; D014771:Virion / Q000378:metabolism; D019065:Virus Assembly
D012357:RNA, Transfer, Lys; D008250:Lysine-tRNA Ligase