Characterization of HIV-1 REV protein: binding stoichiometry and minimal RNA substrate.
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.
Cook
K S
KS
Repligen Corporation, Cambridge, MA 02139.
Fisk
G J
GJ
Hauber
J
J
Usman
N
N
Daly
T J
TJ
Rusche
J R
JR
D001345:Autoradiography; D001483:Base Sequence; D015694:Gene Products, rev / Q000737:chemistry* / Q000378:metabolism; D005814:Genes, Viral; D015752:Genes, env; D015497:HIV-1 / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D012367:RNA, Viral / Q000737:chemistry* / Q000378:metabolism; D054321:rev Gene Products, Human Immunodeficiency Virus
D015694:Gene Products, rev; D012367:RNA, Viral; D054321:rev Gene Products, Human Immunodeficiency Virus