Profiling non-lysyl tRNAs in HIV-1

  1. Mariana Pavon-Eternod1,5,
  2. Min Wei2,3,5,
  3. Tao Pan1 and
  4. Lawrence Kleiman2,3,4
  1. 1Department of Biochemistry and Molecular Biology, University of Chicago, Chicago, Illinois 60637, USA
  2. 2Lady Davis Institute for Medical Research and McGill AIDS Centre, Jewish General Hospital, Montreal, Quebec H3T1E2, Canada
  3. 3Department of Medicine, McGill University, Montreal, Quebec H3T1E2, Canada
  4. 4Department of Microbiology and Immunology, McGill University, Montreal, Quebec H3T1E2, Canada

  1. ↵5 These authors contributed equally to this work.

Abstract

During its assembly, human HIV-1 selectively packages the tRNALys isoacceptors, including tRNALys3, the primer for the reverse transcriptase. However, other low molecular weight RNA species are also seen in the virus. We profiled the tRNAs packaged into HIV-1 using microarray analysis and validated our results by two-dimensional gel electrophoresis and RT-PCR. In addition to tRNALys isoacceptors, tRNAAsn and the rare isoacceptor of tRNAIle are also selectively packaged. In Gag viral-like particles missing the GagPol protein, overall tRNA incorporation is reduced by >80%. This reduction is significantly greater than can be accounted for by the reduction in tRNALys isoacceptors, tRNAAsn and tRNAIle, suggesting that incorporation of other tRNAs may also require the GagPol protein. These results demonstrate selective incorporation of non-lysyl tRNAs into HIV-1 and highlight the application of microarrays as a novel method to study tRNA incorporation into viruses.

Keywords:

Keywords

Footnotes

  • Reprint requests to: Tao Pan, Department of Biochemistry and Molecular Biology, University of Chicago, Chicago, IL 60637, USA; e-mail: taopan{at}uchicago.edu; fax: (773) 702-0439; or Lawrence Kleiman, Lady Davis Institute for Medical Research and McGill AIDS Centre, Jewish General Hospital, Montreal, Quebec H3T1E2, Canada; e-mail: lawrence.kleiman{at}mcgill.ca; fax: (514) 340-8260.

  • Article published online ahead of print. Article and publication date are at http://www.rnajournal.org/cgi/doi/10.1261/rna.1928110.

    • Received September 14, 2009.
    • Accepted October 20, 2009.

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  1. Published in Advance December 9, 2009, doi: 10.1261/rna.1928110 RNA 16: 267-273 Copyright © 2010 RNA Society

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