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Structural and functional analysis of the visna virus Rev-response element
1Howard Hughes Medical Institute, Duke University Medical Center, Durham, North Carolina 27710.
Journal of Virology
|June 1, 1992
Summary
Visna virus and human immunodeficiency virus type 1 (HIV-1) Rev proteins regulate viral gene expression. Though functionally similar, these Rev proteins exhibit distinct RNA binding specificities, explaining their homologous viral system function.
Area of Science:
- Virology
- Molecular Biology
- RNA Biology
Background:
- Human immunodeficiency virus type 1 (HIV-1) and visna virus are distantly related lentiviruses.
- Both viruses encode a Rev protein, a posttranscriptional regulator essential for viral structural protein expression.
Purpose of the Study:
- To genetically map and characterize the cis-acting target sequence for visna virus Rev (RRE-V).
- To investigate the RNA binding specificities of visna virus Rev and HIV-1 Rev.
Main Methods:
- Genetic mapping of the visna virus Rev-response element (RRE-V).
- Computer-predicted RNA structure analysis of RRE-V.
- In vitro validation using structure-specific RNase cleavage assays.
- In vitro binding assays to assess Rev protein-RRE interactions.
Main Results:
- The RRE-V was mapped to a 176-nucleotide RNA stem-loop structure.
- Visna virus Rev specifically bound RRE-V in vitro, but not the HIV-1 RRE.
- HIV-1 Rev did not specifically bind RRE-V.
Conclusions:
- Visna virus and HIV-1 Rev proteins, despite functional analogy, possess distinct RNA sequence specificities.
- These differing specificities provide a biochemical basis for the observation that Rev proteins function only within their homologous viral systems.