title: RNA recognition by the joint action of two nucleolin RNA-binding domains: genetic analysis and structural modeling.
abstract: The interaction of nucleolin with a short stem-loop structure (NRE) requires two contiguous RNA-binding domains (RBD 1+2). The structural basis for RNA recognition by these RBDs was studied using a genetic system in Escherichia coli. Within each of the two domains, we identified several mutations that severely impair interaction with the RNA target. Mutations that alter RNA-binding specificity were also isolated, suggesting the identity of specific contacts between RBD 1+2 amino acids and nucleotides within the NRE stem-loop. Our data indicate that both RBDs participate in a joint interaction with the NRE and that each domain uses a different surface to contact the RNA. The constraints provided by these genetic data and previous mutational studies have enabled us to propose a three-dimensional model of nucleolin RBD 1+2 bound to the NRE stem-loop.
authors.affiliation: Laboratoire de Biologie Moléculaire Eucaryote, Institut de Biologie Cellulaire et de Génétique du CNRS, UPR 9006, 118 route de Narbonne, 31062 Toulouse Cedex, France. Bouvet@ibcg.biotoul.fr
authors.forename: P
authors.initials: P
authors.lastname: Bouvet
authors.forename: C
authors.initials: C
authors.lastname: Jain
authors.forename: J G
authors.initials: JG
authors.lastname: Belasco
authors.forename: F
authors.initials: F
authors.lastname: Amalric
authors.forename: M
authors.initials: M
authors.lastname: Erard
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D004252:DNA Mutational Analysis; D004926:Escherichia coli / Q000235:genetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D010446:Peptide Fragments / Q000235:genetics / Q000378:metabolism; D010750:Phosphoproteins / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding / Q000235:genetics; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D013489:Suppression, Genetic; D000097796:Nucleolin
chemical_list: D010446:Peptide Fragments; D010750:Phosphoproteins; D016601:RNA-Binding Proteins; D011994:Recombinant Proteins; D000097796:Nucleolin