Journal
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
Volume 117, Issue 28, Pages 16333-16338Publisher
NATL ACAD SCIENCES
DOI: 10.1073/pnas.2004170117
Keywords
ribosome; tRNA; anticodon; miscoding; structural biology
Categories
Funding
- NIH [R01 GM093278]
- National Institute of General Medical Sciences (NIGMS) [P30 GM124165, RR029205, OD021527]
- SER-CAT beamlines (member institutions and NIH Equipment) [S10_RR25528, S10_RR028976]
- US Department of Energy Office of Science User Facility operated by Argonne National Laboratory [DE-AC02-06CH11357, W-31-109-Eng-38]
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Bacterial transfer RNAs (tRNAs) contain evolutionarily conserved sequences and modifications that ensure uniform binding to the ribosome and optimal translational accuracy despite differences in their aminoacyl attachments and anticodon nucleotide sequences. In the tRNA anticodon stem-loop, the anticodon sequence is cor-related with a base pair in the anticodon loop (nucleotides 32 and 38) to tune the binding of each tRNA to the decoding center in the ribosome. Disruption of this correlation renders the ribosome un-able to distinguish correct from incorrect tRNAs. The molecular basis for how these two tRNA features combine to ensure accurate decoding is unclear. Here, we solved structures of the bacterial ribosome containing either wild-type tRNA(GGC)(Ala) or tRNA(GGC)(Ala) contain-ing a reversed 32-38 pair on cognate and near-cognate codons. Structures of wild-type tRNA(GGC)(Ala) bound to the ribosome reveal 23S ribosomal RNA (rRNA) nucleotide A1913 positional changes that are dependent on whether the codon-anticodon interaction is cognate or near cognate. Further, the 32-38 pair is destabilized in the context of a near-cognate codon-anticodon pair. Reversal of the pairing in tRNA(GGC)(Ala) ablates A1913 movement regardless of whether the interaction is cognate or near cognate. These results demonstrate that disrupting 32-38 and anticodon sequences al-ters interactions with the ribosome that directly contribute to misreading.
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