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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
PubMed
Summary
This summary is machine-generated.

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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.

Related Experiment Videos

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.