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Journal of Virology, August 2001, p. 6835-6840, Vol. 75, No. 15
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.15.6835-6840.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Gag-Pol Supplied in trans Is Efficiently Packaged and Supports Viral Function in Human Immunodeficiency Virus Type 1

M. K. Hill,1 C. W. Hooker,2 D. Harrich,2 S. M. Crowe,1 and J. Mak1,*

AIDS Pathogenesis Research Unit, Macfarlane Burnet Centre for Medical Research, Fairfield, Victoria 3078,1 and Sir Albert Sakzewski Virus Research Centre, Royal Children's Hospital, Herston, Queensland 4029,2 Australia

Received 18 January 2001/Accepted 12 April 2001

The intracellular trafficking and subsequent incorporation of Gag-Pol into human immunodeficiency virus type 1 (HIV-1) remains poorly defined. Gag-Pol is encoded by the same mRNA as Gag and is generated by ribosomal frameshifting. The multimerization of Gag and Gag-Pol is an essential step in the formation of infectious viral particles. In this study, we examined whether the interaction between Gag and Gag-Pol is initiated during protein translation in order to facilitate the trafficking and subsequent packaging of Gag-Pol into the virion. A conditional cotransfection system was developed in which virion formation required the coexpression of two HIV-1-based plasmids, one that produces both Gag and Gag-Pol and one that only produces Gag-Pol. The Gag-Pol proteins were either immunotagged with a His epitope or functionally tagged with a mutation (K65R) in reverse transcriptase that is associated with drug resistance. Gag-Pol packaging was assessed to determine whether the Gag-Pol incorporated into the virion was preferentially packaged from the plasmid that expressed both Gag and Gag-Pol or whether it could be packaged from either plasmid. Our data show that translation of Gag and Gag-Pol from the same mRNA is not critical for virion packaging of the Gag-Pol polyprotein or for viral function.

* Corresponding author. Mailing address: The Macfarlane Burnet Centre for Medical Research, P.O. Box 254, Fairfield, Victoria, Australia 3078. Phone: 61 3 9282 2217. Fax: 61 3 9482 6152. E-mail: mak{at}burnet.edu.au.

Journal of Virology, August 2001, p. 6835-6840, Vol. 75, No. 15
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.15.6835-6840.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Aguiar, R. S., Pereira, H. S., Costa, L. J., Brindeiro, R. M., Tanuri, A. (2006). Gag-Pol bearing a reverse transcriptase drug-resistant mutation influences viral genomic RNA incorporation into human immunodeficiency virus type 1 particles. J. Gen. Virol. 87: 2669-2677 [Abstract] [Full Text]  
  • Apolloni, A., Hooker, C. W., Mak, J., Harrich, D. (2003). Human Immunodeficiency Virus Type 1 Protease Regulation of Tat Activity Is Essential for Efficient Reverse Transcription and Replication. J. Virol. 77: 9912-9921 [Abstract] [Full Text]  
  • Hill, M. K., Shehu-Xhilaga, M., Campbell, S. M., Poumbourios, P., Crowe, S. M., Mak, J. (2003). The Dimer Initiation Sequence Stem-Loop of Human Immunodeficiency Virus Type 1 Is Dispensable for Viral Replication in Peripheral Blood Mononuclear Cells. J. Virol. 77: 8329-8335 [Abstract] [Full Text]  
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