title: RNA binding site of R17 coat protein.
abstract: The specific interaction between R17 coat protein and its target of translational repression at the initiation site of the R17 replicase gene was studied by synthesizing variants of the RNA binding site and measuring their affinity to the coat protein by using a nitrocellulose filter binding assay. Substitution of two of the seven single-stranded residues by other nucleotides greatly reduced the Ka, indicating that they are essential for the RNA-protein interaction. In contrast, three other single-stranded residues can be substituted without altering the Ka. When several of the base-paired residues in the binding site are altered in such a way that pairing is maintained, little change in Ka is observed. However, when the base pairs are disrupted, coat protein does not bind. These data suggest that while the hairpin loop structure is essential for protein binding, the base-paired residues do not contact the protein directly. On the basis of these and previous data, a model for the structural requirements of the R17 coat protein binding site is proposed. The model was successfully tested by demonstrating that oligomers with sequences quite different from the replicase initiator were able to bind coat protein.
authors.forename: P J
authors.initials: PJ
authors.lastname: Romaniuk
authors.forename: P
authors.initials: P
authors.lastname: Lowary
authors.forename: H N
authors.initials: HN
authors.lastname: Wu
authors.forename: G
authors.initials: G
authors.lastname: Stormo
authors.forename: O C
authors.initials: OC
authors.lastname: Uhlenbeck
mesh_terms: D001482:Base Composition; D001665:Binding Sites; D002213:Capsid / Q000378:metabolism*; D036022:Capsid Proteins; D003090:Coliphages / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism; D007202:Indicators and Reagents; D007700:Kinetics; D009843:Oligoribonucleotides / Q000138:chemical synthesis; D011485:Protein Binding; D012367:RNA, Viral / Q000378:metabolism*; D016601:RNA-Binding Proteins; D013329:Structure-Activity Relationship
chemical_list: D036022:Capsid Proteins; D007202:Indicators and Reagents; D009843:Oligoribonucleotides; D012367:RNA, Viral; D016601:RNA-Binding Proteins