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Nucleic Acids Research, 1991, Vol. 19, No. 7 1577-1583
© 1991

MOLECULAR BIOLOGY

Characterization of HIV-1 REV protein: binding stoichiometry and minimal RNA substrate

Kathleen Sue Cook+, Gregory J. Fisk, Joachim Hauber1, Nassim Usman2, Thomas J. Daly and James R. Rusche*

Repligen Corporation One Kendall Square, Building 700, Cambridge, MA 02139, USA 1Sandoz Research Institute Vienna, Austria 2Massachusetts Institute of Technology Cambridge, MA 02139, USA

* To whom correspondence should be addressed

Received December 6, 1990. Revised February 19, 1991. Accepted February 19, 1991.

The HIV-1 REV protein binds to the stem II region of the REV-responsive element (RNA). Studies to further define the RNA sequence and structure specifically bound by REV protein identify a minimal RNA element of 40 nucleotides. Analysis of RNA fragments by gel retardation and filter binding suggest that a core element composed of one particular stem with flanking sequences capable of forming a second double stranded region is essential for specific recognition by REV protein. Stable REV-RNA complexes are formed in a stoichiometry of 1 REV: 1 RNA. The minimal RNA element binds 1 REV molecule while the stem II saturates at 3 REV molecules per RNA. These results establish that REV recognizes a primary binding site within the RRE and support the notion that the initial viral transcript binding event involves a monomeric REV protein.

+ Present address: Tufts University, Medford, MA 02145, USA


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