title: Cooperative dimerization of a stably folded protein directed by a flexible RNA in the assembly of the HIV Rev dimer-RRE stem II complex.
abstract: The binding of the HIV-1 Rev protein as an oligomer to a viral RNA element, the Rev-response element (RRE), mediates nuclear export of genomic RNA. Assembly of the Rev-RRE ribonucleoprotein (RNP) complex is nucleated by the binding of the first Rev molecule to stem IIB of the RRE. This is followed by stepwise addition of a total of ~six Rev molecules along the RRE through a combination of RNA-protein and protein-protein interactions. RRE stem II, which forms a three-way junction consisting of stems IIA, IIB and IIC, has been shown to bind to two Rev molecules in a cooperative manner, with the second Rev molecule binding to the junction region of stem II. The results of base substitutions at the stem II junction, and characterization of stem II junction variants selected from a randomized library showed that an "open" flexible structure is preferred for binding of the second Rev molecule, and that binding of the second Rev molecule to the junction region is not sequence-specific. Alanine substitutions of a number of Rev amino acid residues implicated to be important for Rev folding in previous structural studies were found to result in a dramatic decrease in the binding of the second Rev molecule. These results support the model that proper folding of Rev is critical in ensuring that the flexible RRE is able to correctly position Rev molecules for specific RNP assembly, and suggests that targeting Rev folding may be effective in the inhibition of Rev function.
authors.affiliation: Department of Life Sciences, Tokyo Gakugei University, Koganei, Tokyo, 184-8501, Japan.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Tanamura
authors.affiliation: Department of Life Sciences, Tokyo Gakugei University, Koganei, Tokyo, 184-8501, Japan.
authors.forename: Hiroto
authors.initials: H
authors.lastname: Terakado
authors.affiliation: Department of Life Sciences, Tokyo Gakugei University, Koganei, Tokyo, 184-8501, Japan.
authors.forename: Kazuo
authors.initials: K
authors.lastname: Harada
mesh_terms: D019943:Amino Acid Substitution; D015497:HIV-1 / Q000235:genetics* / Q000378:metabolism; D008958:Models, Molecular; D009690:Nucleic Acid Conformation; D017510:Protein Folding; D055503:Protein Multimerization; D012367:RNA, Viral / Q000737:chemistry / Q000378:metabolism*; D020218:Response Elements; D012261:Ribonucleoproteins / Q000378:metabolism; D054321:rev Gene Products, Human Immunodeficiency Virus / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
chemical_list: D012367:RNA, Viral; D012261:Ribonucleoproteins; D054321:rev Gene Products, Human Immunodeficiency Virus; C514860:rev protein, Human Immunodeficiency Virus-1
keywords: HIV Rev-RRE complex; RNA structure; RNA-protein interactions; cooperativity; protein folding