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Journal of Virology, February 2004, p. 1595-1601, Vol. 78, No. 3
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.3.1595-1601.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Ability of Wild-Type and Mutant Lysyl-tRNA Synthetase To Facilitate tRNALys Incorporation into Human Immunodeficiency Virus Type 1

Shan Cen,1,{dagger} Hassan Javanbakht,1,2,{dagger} Meijuan Niu,1 and Lawrence Kleiman1,2,3*

Lady Davis Institute for Medical Research and McGill AIDS Centre, Jewish General Hospital,1 Departments of Medicine,2 Microbiology and Immunology, McGill University, Montreal, Quebec, Canada H3T 1E23

Received 20 June 2003/ Accepted 17 October 2003

The major human tRNALys 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 tRNALys, via its interaction with Gag, and it is a strong candidate for being the signal that specifically targets tRNALys 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 tRNALys. Herein, we show that this increase in tRNALys incorporation into virions is dependent upon the ability of LysRS to bind to tRNALys but not upon its ability to aminoacylate the tRNALys. 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 tRNALys, does not increase tRNALys packaging into viruses, while C-terminally truncated LysRS, which binds to but does not aminoacylate tRNALys, still facilitates an increase in tRNALys packaging into virions.


* Corresponding author. Mailing address: McGill AIDS Centre, Lady Davis Institute—Jewish General Hospital, 3755 Cote St. Catherine Rd., Montreal, Quebec, Canada H3T 1E2. Phone: (514) 340-8260. Fax: (514) 340-7502. E-mail: lawrence.kleiman{at}mcgill.ca.

{dagger} S.C. and H.J. contributed equally to this report.


Journal of Virology, February 2004, p. 1595-1601, Vol. 78, No. 3
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.3.1595-1601.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.




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