title: The role of a clinically important mutation in the fold and RNA-binding properties of KH motifs.
abstract: We have investigated the role in the fold and RNA-binding properties of the KH modules of a hydrophobic to asparagine mutation of clinical importance in the fragile X syndrome. The mutation involves a well-conserved hydrophobic residue close to the N terminus of the second helix of the KH fold (alpha2(3) position). The effect of the mutation has been long debated: Although the mutant has been shown to disrupt the three-dimensional fold of several KH domains, the residue seems also to be directly involved in RNA binding, the main function of the KH module. Here we have used the KH3 of Nova-1, whose structure is known both in isolation and in an RNA complex, to study in detail the role of the alpha2(3) position. A detailed comparison of Nova KH3 structure with its RNA/KH complex and with other KH structures suggests a dual role for the alpha2(3) residue, which is involved both in stabilizing the hydrophobic core and in RNA contacts. We further show by nuclear magnetic resonance (NMR) studies in solution that L447 of Nova-1 in position alpha2(3) is in exchange in the absence of RNA, and becomes locked in a more rigid conformation only upon formation of an RNA complex. This implies that position alpha2(3) functions as a "gate" in the mechanism of RNA recognition of KH motifs based on the rigidification of the fold upon RNA binding.
authors.affiliation: National Institute for Medical Research, London NW7 1AA, UK.
authors.forename: Andres
authors.initials: A
authors.lastname: Ramos
authors.forename: David
authors.initials: D
authors.lastname: Hollingworth
authors.forename: Annalisa
authors.initials: A
authors.lastname: Pastore
mesh_terms: D020816:Amino Acid Motifs / Q000502:physiology*; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D011485:Protein Binding / Q000502:physiology; D017510:Protein Folding; D012313:RNA / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000737:chemistry
chemical_list: D016601:RNA-Binding Proteins; D012313:RNA