A functional interaction between ribosomal proteins S7 and S11 within the bacterial ribosome.
In this study, we used site-directed mutagenesis to disrupt an interaction that had been detected between ribosomal proteins S7 and S11 in the crystal structure of the bacterial 30 S subunit. This interaction, which is located in the E site, connects the head of the 30 S subunit to the platform and is involved in the formation of the exit channel through which passes the 30 S-bound messenger RNA. Neither mutations in S7 nor mutations in S11 prevented the incorporation of the proteins into the 30 S subunits but they perturbed the function of the ribosome. In vivo assays showed that ribosomes with either mutated S7 or S11 were altered in the control of translational fidelity, having an increased capacity for frameshifting, readthrough of a nonsense codon and codon misreading. Toeprinting and filter-binding assays showed that 30 S subunits with either mutated S7 or S11 have an enhanced capacity to bind mRNA. The effects of the S7 and S11 mutations can be related to an increased flexibility of the head of the 30 S, to an opening of the mRNA exit channel and to a perturbation of the proposed allosteric coupling between the A and E sites. Altogether, our results demonstrate that S7 and S11 interact in a functional manner and support the notion that protein-protein interactions contribute to the dynamics of the ribosome.
Robert
Francis
F
Département de Biochimie, Université de Montréal, Montréal, Québec H3C 3J7, Canada.
Brakier-Gingras
Léa
L
D000595:Amino Acid Sequence; D001665:Binding Sites; D015153:Blotting, Western; D018389:Codon, Nonsense; D003460:Crystallization; D004347:Drug Interactions; D004926:Escherichia coli / Q000737:chemistry / Q000235:genetics / Q000648:ultrastructure*; D016368:Frameshift Mutation; D015870:Gene Expression; D008156:Luciferases / Q000235:genetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D015394:Molecular Structure; D016297:Mutagenesis, Site-Directed; D014176:Protein Biosynthesis / Q000502:physiology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012343:RNA, Transfer / Q000378:metabolism; D011993:Recombinant Fusion Proteins; D012269:Ribosomal Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D012270:Ribosomes / Q000737:chemistry* / Q000502:physiology / Q000648:ultrastructure; D016415:Sequence Alignment; D013329:Structure-Activity Relationship
D018389:Codon, Nonsense; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D012269:Ribosomal Proteins; C045454:ribosomal protein S11; C037442:ribosomal protein S7; D012343:RNA, Transfer; D008156:Luciferases