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Journal of Virology, June 2000, p. 5441-5451, Vol. 74, No. 12
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

The Double-Stranded RNA-Binding Protein Staufen Is Incorporated in Human Immunodeficiency Virus Type 1: Evidence for a Role in Genomic RNA Encapsidation

Andrew J. Mouland,1 Johanne Mercier,1 Ming Luo,2 Luc Bernier,1 Luc DesGroseillers,2 and Éric A. Cohen1,*

Departments of Microbiology & Immunology1 and Biochemistry,2 Université de Montréal, Montreal, Quebec H3C 3J7, Canada

Received 6 December 1999/Accepted 21 March 2000

Human Staufen (hStau), a double-stranded RNA (dsRNA)-binding protein that is involved in mRNA transport, is incorporated in human immunodeficiency virus type 1 (HIV-1) and in other retroviruses, including HIV-2 and Moloney murine leukemia virus. Sucrose and Optiprep gradient analyses reveal cosedimentation of hStau with purified HIV-1, while subtilisin assays demonstrate that it is internalized. hStau incorporation in HIV-1 is selective, is dependent on an intact functional dsRNA-binding domain, and quantitatively correlates with levels of encapsidated HIV-1 genomic RNA. By coimmunoprecipitation and reverse transcription-PCR analyses, we demonstrate that hStau is associated with HIV-1 genomic RNA in HIV-1-expressing cells and purified virus. Overexpression of hStau enhances virion incorporation levels, and a corresponding, threefold increase in HIV-1 genomic RNA encapsidation levels. This coordinated increase in hStau and genomic RNA packaging had a significant negative effect on viral infectivity. This study is the first to describe hStau within HIV-1 particles and provides evidence that hStau binds HIV-1 genomic RNA, indicating that it may be implicated in retroviral genome selection and packaging into assembling virions.

* Corresponding author. Mailing address: Human Retrovirology Laboratory, Université de Montréal, P.O. Box 6128, Station Centreville, Montreal, Quebec, Canada H3C 3J7. Phone: (514) 343-5967. Fax: (514) 343-5995. E-mail: eric.cohen{at}umontreal.ca.

Journal of Virology, June 2000, p. 5441-5451, Vol. 74, No. 12
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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