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05:46
Rapid Screening of HIV Reverse Transcriptase and Integrase Inhibitors
Published on: April 9, 2014
HuR interacts with human immunodeficiency virus type 1 reverse transcriptase, and modulates reverse transcription in
Julie Lemay1, Priscilla Maidou-Peindara, Thomas Bader
1Institut Cochin, Université Paris Descartes, CNRS (UMR8104), Paris, France. julie.lemay@kispi.uzh.ch
Retrovirology
|June 12, 2008
Summary
The RNA-binding protein HuR interacts with HIV-1 reverse transcriptase, impacting viral replication. Silencing HuR inhibits human immunodeficiency virus type 1 (HIV-1) infection, suggesting HuR modulates reverse transcription.
Area of Science:
- Virology
- Molecular Biology
- Cellular Biology
Background:
- Reverse transcription is a crucial step in human immunodeficiency virus type 1 (HIV-1) replication, catalyzed by reverse transcriptase (RT).
- The cellular proteins interacting with the HIV-1 reverse transcription complex (RTC) remain largely uncharacterized.
- The role of cellular proteins in modulating viral replication is an active area of research.
Purpose of the Study:
- To investigate the interaction between cellular proteins and HIV-1 reverse transcriptase (RT).
- To determine the effect of the nucleocytoplasmic shuttling protein HuR on HIV-1 reverse transcription and infection.
Main Methods:
- Yeast two-hybrid screening to identify protein-protein interactions.
- Homogenous time-resolved fluorescence (HTRF) for in vitro interaction confirmation.
- RNA interference (siRNA) for HuR silencing.
- Site-directed mutagenesis and immunoprecipitation studies.
Main Results:
- A direct protein-protein interaction was identified between HIV-1 RT and the RNA-binding protein HuR.
- HuR binds to the RNase H domain of RT and the C-terminus of HuR.
- Silencing HuR significantly impaired HIV-1 reverse transcription and infection.
- HuR binding to viral RNA was not detected, suggesting a protein-protein interaction mechanism.
Conclusions:
- The RNA-binding protein HuR interacts with HIV-1 RT via protein-protein interaction.
- HuR plays a significant role in modulating HIV-1 reverse transcription and infection.
- HuR's mechanism of action involves interaction with RT, not viral RNA, suggesting a novel regulatory pathway in HIV-1 replication.
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