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Sam68 is absolutely required for Rev function and HIV-1 production.
Suhasini Modem1, Kameswara R Badri, Thomas C Holland
1Department of Immunology and Microbiology, Wayne State University-School of Medicine Detroit, MI 48201, USA.
Nucleic Acids Research
|February 11, 2005
Summary
Sam68 protein is essential for HIV-1 production and RNA export. Knockdown of Sam68 (small nuclear ribonucleoprotein-binding protein) inhibits viral replication by blocking Rev-mediated export of viral RNA.
Area of Science:
- Molecular Virology
- Cell Biology
- RNA Biology
Background:
- HIV-1 Rev protein is crucial for viral gene expression and replication.
- Sam68 (Src-associated protein 68) interacts with HIV-1 Rev, but its precise role is unclear.
- Sam68's C-terminal mutants negatively impact Rev function and HIV-1 production.
Purpose of the Study:
- To elucidate the mechanism of Sam68 in Rev/RRE function.
- To investigate the necessity of Sam68 for HIV-1 production and RNA export.
Main Methods:
- Utilized RNA interference (RNAi) to create stable Sam68 knockdown HeLa cells (SSKH).
- Assessed Rev-mediated reporter gene expression (CAT and gag).
- Quantified HIV-1 production and viral mRNA export to the cytoplasm.
Main Results:
- Sam68 knockdown abrogated Rev-mediated reporter gene expression.
- HIV-1 production was dramatically inhibited in SSKH cells.
- Defective Rev-mediated RNA export of RRE-containing and unspliced viral mRNAs was observed.
Conclusions:
- Sam68 plays a critical role in Rev-mediated RNA export.
- Sam68 is indispensable for efficient HIV-1 production.