title: Structure and RNA binding of the third KH domain of poly(C)-binding protein 1.
abstract: Poly(C)-binding proteins (CPs) are important regulators of mRNA stability and translational regulation. They recognize C-rich RNA through their triple KH (hn RNP K homology) domain structures and are thought to carry out their function though direct protection of mRNA sites as well as through interactions with other RNA-binding proteins. We report the crystallographically derived structure of the third domain of alphaCP1 to 2.1 A resolution. alphaCP1-KH3 assumes a classical type I KH domain fold with a triple-stranded beta-sheet held against a three-helix cluster in a betaalphaalphabetabetaalpha configuration. Its binding affinity to an RNA sequence from the 3'-untranslated region (3'-UTR) of androgen receptor mRNA was determined using surface plasmon resonance, giving a K(d) of 4.37 microM, which is indicative of intermediate binding. A model of alphaCP1-KH3 with poly(C)-RNA was generated by homology to a recently reported RNA-bound KH domain structure and suggests the molecular basis for oligonucleotide binding and poly(C)-RNA specificity.
authors.affiliation: School of Biomedical and Chemical Sciences, the UWA Centre for Medical Research, The University of Western Australia WA Australia 6009.
authors.forename: M
authors.initials: M
authors.lastname: Sidiqi
authors.forename: J A
authors.initials: JA
authors.lastname: Wilce
authors.forename: J P
authors.initials: JP
authors.lastname: Vivian
authors.forename: C J
authors.initials: CJ
authors.lastname: Porter
authors.forename: A
authors.initials: A
authors.lastname: Barker
authors.forename: P J
authors.initials: PJ
authors.lastname: Leedman
authors.forename: M C J
authors.initials: MC
authors.lastname: Wilce
mesh_terms: D020413:3' Untranslated Regions / Q000737:chemistry* / Q000378:metabolism; D000595:Amino Acid Sequence; D001665:Binding Sites; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009841:Oligonucleotides / Q000737:chemistry; D011066:Poly C / Q000737:chemistry; D017434:Protein Structure, Tertiary; D016601:RNA-Binding Proteins / Q000737:chemistry* / Q000378:metabolism; D016415:Sequence Alignment; D020349:Surface Plasmon Resonance
chemical_list: D020413:3' Untranslated Regions; D009841:Oligonucleotides; D016601:RNA-Binding Proteins; D011066:Poly C
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