Letter abstract
Nature Chemical Biology 2, 254 - 258 (2006)
Published online: 2 April 2006 | doi:10.1038/nchembio783
Functional epitopes at the ribosome subunit interface
Oliver Rackham1,2, Kaihang Wang1,2 & Jason W Chin1
The ribosome is a 2.5-MDa molecular machine that synthesizes cellular proteins encoded in mRNAs1. The 30S and 50S subunits of the ribosome associate through structurally defined intersubunit bridges2, 3, 4, 5 burying 6,000 Å2, 80% of which is buried in conserved RNA-RNA interactions6. Intersubunit bridges bind translation factors7, 8, may coordinate peptide bond formation and translocation9, 10, 11 and may be actively remodeled in the post-termination complex12, 13, but the functional importance of numerous 30S bridge nucleotides had been unknown. We carried out large-scale combinatorial mutagenesis and in vivo selections on 30S nucleotides that form RNA-RNA intersubunit bridges in the Escherichia coli ribosome. We determined the covariation and functional importance of bridge nucleotides, allowing comparison of the structural interface and phylogenetic data to the functional epitope. Our results reveal how information for ribosome function is partitioned across bridges, and suggest a subset of nucleotides that may have measurable effects on individual steps of the translational cycle.
- Medical Research Council Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, England, UK.
- These authors contributed equally to this work.
Correspondence to: Jason W Chin1 e-mail: chin@mrc-lmb.cam.ac.uk
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