Important note: Information in this article was accurate in 2000. The state of the art may have changed since the publication date.
The double-stranded RNA-binding protein Staufen is incorporated in human immunodeficiency virus type 1: evidence for a role in genomic RNA encapsidation.
Abstract:
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.
Keywords: JOURNAL ARTICLE Binding Sites Cell Line Centrifugation, Density Gradient Cloning, Molecular Gene Expression Gene Products, gag/GENETICS/METABOLISM *Genome, Viral Human HIV-1/CHEMISTRY/GENETICS/*METABOLISM/PATHOGENICITY HIV-2/CHEMISTRY/METABOLISM Moloney Leukemia Virus/CHEMISTRY/METABOLISM Mutation/GENETICS Precipitin Tests Protein Precursors/GENETICS/METABOLISM Reverse Transcriptase Polymerase Chain Reaction RNA-Binding Proteins/ANALYSIS/CHEMISTRY/GENETICS/*METABOLISM RNA, Double-Stranded/GENETICS/*METABOLISM RNA, Viral/ANALYSIS/GENETICS/*METABOLISM Substrate Specificity Subtilisin/METABOLISM Support, Non-U.S. Gov't Transfection *Virus Assembly
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Important note: Information in this article was accurate in 2000. The state of the art may have changed since the publication date.
