Specific Inhibition of the Synthesis of Human Lysyl-tRNA Synthetase Results in Decreases in tRNALys Incorporation, tRNA3LysAnnealing to Viral RNA, and Viral …

F Guo, S Cen, M Niu, H Javanbakht… - Journal of virology, 2003 - Am Soc Microbiol
F Guo, S Cen, M Niu, H Javanbakht, L Kleiman
Journal of virology, 2003Am Soc Microbiol
The major human tRNALys isoacceptors, and, are selectively packaged into human
immunodeficiency virus type 1 (HIV-1) during assembly, where 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 is a strong candidate for being the signal that
specifically targets tRNALys for viral incorporation. We have transfected 293T cells with HIV-
1 proviral DNA and short interfering RNA (siRNA) specific for LysRS to study the effect of …
Abstract
The major human tRNALys isoacceptors, \batchmode \documentclass[fleqn,10pt,legalpaper]{article} \usepackage{amssymb} \usepackage{amsfonts} \usepackage{amsmath} \pagestyle{empty} \begin{document} \(tRNA_{1,2}^{Lys}\) \end{document} and \batchmode \documentclass[fleqn,10pt,legalpaper]{article} \usepackage{amssymb} \usepackage{amsfonts} \usepackage{amsmath} \pagestyle{empty} \begin{document} \(tRNA_{3}^{Lys}\) \end{document}, are selectively packaged into human immunodeficiency virus type 1 (HIV-1) during assembly, where \batchmode \documentclass[fleqn,10pt,legalpaper]{article} \usepackage{amssymb} \usepackage{amsfonts} \usepackage{amsmath} \pagestyle{empty} \begin{document} \(tRNA_{3}^{Lys}\) \end{document} 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 is a strong candidate for being the signal that specifically targets tRNALys for viral incorporation. We have transfected 293T cells with HIV-1 proviral DNA and short interfering RNA (siRNA) specific for LysRS to study the effect of diminished cellular LysRS upon tRNALys packaging, \batchmode \documentclass[fleqn,10pt,legalpaper]{article} \usepackage{amssymb} \usepackage{amsfonts} \usepackage{amsmath} \pagestyle{empty} \begin{document} \(tRNA_{3}^{Lys}\) \end{document} annealing to viral genomic RNA, and viral production and infectivity. At early time points after siRNA transfection, an 80% inhibition of LysRS incorporation into viruses reflects an 80% reduction of newly synthesized LysRS, rather than a more limited 20 to 25% decrease in the concentration of total cell LysRS, indicating that newly synthesized LysRS in the cell may be the main source of viral LysRS. Viruses produced from cells transfected with siRNA show reduced tRNALys packaging, reduced \batchmode \documentclass[fleqn,10pt,legalpaper]{article} \usepackage{amssymb} \usepackage{amsfonts} \usepackage{amsmath} \pagestyle{empty} \begin{document} \(tRNA_{3}^{Lys}B\) \end{document} annealing to viral RNA, and reduced viral infectivity.
American Society for Microbiology