Dissecting RNA-protein interactions: RNA-RNA recognition by Rop.
The ColE1 plasmid of E. coli encodes a small RNA-binding protein, Rop, which is involved in the regulation of plasmid copy number. Rop, a 4-helix bundle protein, facilitates sense-antisense RNA pairing by binding to the transiently formed hairpin pairs of RNA I and the complementary RNA II. We have identified the residues of Rop that are involved in RNA recognition. The residues form a narrow stripe down one face of the bundle and are symmetrically arranged, with recognition centered about two phenylalanine residues. Our results suggest that these phenylalanine residues interact with the loop region of the hairpin pair, with additional interactions between eight polar residues and the phosphate backbone. By modifying the identity of residue 14, we have created a variant of Rop that displays altered RNA binding specificity. The results of our studies allow us to present a detailed picture of RNA-protein recognition in a novel model system.
Predki
P F
PF
Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, Connecticut 06511.
Nayak
L M
LM
Gottlieb
M B
MB
Regan
L
L
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D003086:Bacteriocin Plasmids; D001483:Base Sequence; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009690:Nucleic Acid Conformation; D017354:Point Mutation; D017510:Protein Folding; D017433:Protein Structure, Secondary; D012329:RNA, Bacterial / Q000378:metabolism*; D016601:RNA-Binding Proteins
D001426:Bacterial Proteins; D012329:RNA, Bacterial; D016601:RNA-Binding Proteins; C054243:Rop protein, ColE1 plasmid