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Updated: Jun 26, 2025

Identification of Nucleolar Factors During HIV-1 Replication Through Rev Immunoprecipitation and Mass Spectrometry
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Measurement of HIV Rev-Rev Response Element Functional Activity.

Patrick E H Jackson1,2, Godfrey Dzhivhuho3,4, Jing Huang3,4

  • 1Myles H. Thaler Center for AIDS and Human Retrovirus Research, University of Virginia, Charlottesville, VA, USA. Pej9j@uvahealth.org.

Methods in Molecular Biology (Clifton, N.J.)
|May 14, 2024
PubMed
Summary

Human immunodeficiency virus (HIV) export of unspliced RNA requires the Rev-Response Element (RRE) and Rev protein. We developed a high-throughput assay to analyze Rev-RRE functional activity in HIV isolates.

Keywords:
HIVLentivirusLentivirusesRNA traffickingRNA traffickingRevRev response elementRev response element

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Area of Science:

  • Virology
  • Molecular Biology
  • Genetics

Background:

  • Retroviruses, including HIV, face cellular barriers to exporting unspliced viral messenger RNAs (mRNAs) essential for replication.
  • The human immunodeficiency virus (HIV) employs a specific mechanism involving the viral Rev protein and the Rev-Response Element (RRE) RNA structure to facilitate this export.
  • Variations in the sequence and functional efficiency of the Rev-RRE system exist among different HIV primary isolates.

Purpose of the Study:

  • To develop and validate a high-throughput assay system for assessing the functional activity of the HIV Rev-RRE interaction.
  • To enable comparative analysis of Rev-RRE system functionality across diverse HIV primary isolates.

Main Methods:

  • Utilized packageable viral vectors to create a robust assay system.
  • Developed a high-throughput screening method to quantify Rev-RRE functional activity.

Main Results:

  • Successfully established a high-throughput assay for analyzing Rev-RRE functional activity.
  • The assay system is capable of evaluating functional differences in the Rev-RRE system among various HIV isolates.

Conclusions:

  • The developed high-throughput assay provides a valuable tool for studying the functional diversity of the HIV Rev-RRE system.
  • This system can aid in understanding the implications of Rev-RRE variations in HIV replication and pathogenesis.