title: Engineering specificity changes on a RanBP2 zinc finger that binds single-stranded RNA.
abstract: The realization that gene transcription is much more pervasive than previously thought and that many diverse RNA species exist in simple as well as complex organisms has triggered efforts to develop functionalized RNA-binding proteins (RBPs) that have the ability to probe and manipulate RNA function. Previously, we showed that the RanBP2-type zinc finger (ZF) domain is a good candidate for an addressable single-stranded-RNA (ssRNA) binding domain that can recognize ssRNA in a modular and specific manner. In the present study, we successfully engineered a sequence specificity change onto this ZF scaffold by using a combinatorial approach based on phage display. This work constitutes a foundation from which a set of RanBP2 ZFs might be developed that is able to recognize any given RNA sequence.
authors.affiliation: School of Molecular Bioscience, University of Sydney, Sydney, N.S.W 2006 (Australia).
authors.forename: Marylène
authors.initials: M
authors.lastname: Vandevenne
authors.forename: Mitchell R
authors.initials: MR
authors.lastname: O'Connell
authors.forename: Stephanie
authors.initials: S
authors.lastname: Helder
authors.forename: Nicholas E
authors.initials: NE
authors.lastname: Shepherd
authors.forename: Jacqueline M
authors.initials: JM
authors.lastname: Matthews
authors.forename: Ann H
authors.initials: AH
authors.lastname: Kwan
authors.forename: David J
authors.initials: DJ
authors.lastname: Segal
authors.forename: Joel P
authors.initials: JP
authors.lastname: Mackay
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D018832:Molecular Chaperones / Q000737:chemistry*; D008969:Molecular Sequence Data; D028861:Nuclear Pore Complex Proteins / Q000737:chemistry*; D012313:RNA / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000378:metabolism*; D023822:Tissue Engineering; D016335:Zinc Fingers / Q000235:genetics*
chemical_list: D018832:Molecular Chaperones; D028861:Nuclear Pore Complex Proteins; D016601:RNA-Binding Proteins; C094493:ran-binding protein 2; D012313:RNA
keywords: RNA binding; combinatorial chemistry; phage display; protein design; zinc fingers