title: Probing sequence-specific RNA recognition by the bacteriophage MS2 coat protein.
abstract: We present the results of in vitro binding studies aimed at defining the key recognition elements on the MS2 RNA translational operator (TR) essential for complex formation with coat protein. We have used chemically synthesized operators carrying modified functional groups at defined nucleotide positions, which are essential for recognition by the phage coat protein. These experiments have been complemented with modification-binding interference assays. The results confirm that the complexes which form between TR and RNA-free phage capsids, the X-ray structure of which has recently been reported at 3.0 A, are identical to those which form in solution between TR and a single coat protein dimer. There are also effects on operator affinity which cannot be explained simply by the alteration of direct RNA-protein contacts and may reflect changes in the conformational equilibrium of the unliganded operator. The results also provide support for the approach of using modified oligoribonucleotides to investigate the details of RNA-ligand interactions.
authors.affiliation: Department of Genetics, University of Leeds, UK.
authors.forename: P G
authors.initials: PG
authors.lastname: Stockley
authors.forename: N J
authors.initials: NJ
authors.lastname: Stonehouse
authors.forename: J B
authors.initials: JB
authors.lastname: Murray
authors.forename: S T
authors.initials: ST
authors.lastname: Goodman
authors.forename: S J
authors.initials: SJ
authors.lastname: Talbot
authors.forename: C J
authors.initials: CJ
authors.lastname: Adams
authors.forename: L
authors.initials: L
authors.lastname: Liljas
authors.forename: K
authors.initials: K
authors.lastname: Valegård
mesh_terms: D001482:Base Composition; D001483:Base Sequence; D001665:Binding Sites; D002213:Capsid / Q000737:chemistry / Q000378:metabolism*; D036022:Capsid Proteins; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D018360:Crystallography, X-Ray; D006860:Hydrogen Bonding; D046911:Macromolecular Substances; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009876:Operon; D012313:RNA / Q000737:chemistry / Q000378:metabolism*; D016601:RNA-Binding Proteins
chemical_list: D036022:Capsid Proteins; D046911:Macromolecular Substances; D016601:RNA-Binding Proteins; D012313:RNA