tRNA Fluctuations Observed on Stalled Ribosomes Are Suppressed during Ongoing Protein Synthesis

The pretranslocation complex of the ribosome can undergo spontaneous fluctuations of messenger RNA and transfer RNAs (tRNAs) between classical and hybrid states, and occupation of the hybrid tRNA positions has been proposed to precede translocation. The classical and hybrid state tRNA positions have...

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Bibliographic Details
Published in:Biophysical journal 2017-12, Vol.113 (11), p.2326-2335
Main Authors: Jamiolkowski, Ryan M., Chen, Chunlai, Cooperman, Barry S., Goldman, Yale E.
Format: Article
Language:English
Subjects:
Antibiotics
Elongation
Energy consumption
Energy transfer
Fluctuations
Fluorescence
Fluorescence Resonance Energy Transfer
Guanosine triphosphate
Molecules
mRNA
Nucleic Acids and GenomeBiophysics
Peptide Elongation Factor G - metabolism
Protein Biosynthesis
Protein L11
Protein synthesis
Resonance
Ribonucleic acid
Ribosomal protein L11
Ribosomal Proteins - metabolism
Ribosomes
Ribosomes - genetics
Ribosomes - metabolism
RNA
RNA, Transfer - genetics
Translocation
tRNA
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Summary:The pretranslocation complex of the ribosome can undergo spontaneous fluctuations of messenger RNA and transfer RNAs (tRNAs) between classical and hybrid states, and occupation of the hybrid tRNA positions has been proposed to precede translocation. The classical and hybrid state tRNA positions have been extensively characterized when the ribosome is stalled along the messenger RNA by either the absence or delayed addition of elongation factor G (EF-G), or by the presence of antibiotics or GTP analogs that block translocation. However, during multiple ongoing elongation cycles when both EF-G and ternary complexes are present, EF-G can bind to the pretranslocation complex much faster than the timescale of the classic-hybrid transitions. Using single-molecule fluorescence resonance energy transfer between adjacent tRNAs and between A-site tRNA and ribosomal protein L11, we found that the tRNAs do not fluctuate between the hybrid and classical states, but instead adopt a position with fluorescence resonance energy transfer efficiencies between those of the stalled classical and hybrid states.
ISSN:0006-3495
1542-0086
DOI:10.1016/j.bpj.2017.08.052