Altered tRNA dynamics during translocation on slippery mRNA as determinant of spontaneous ribosome frameshifting

Panagiotis Poulis, Anoshi Patel, Marina V. Rodnina and Sarah Adio ()
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Panagiotis Poulis: Max Planck Institute for Multidisciplinary Sciences
Anoshi Patel: Max Planck Institute for Multidisciplinary Sciences
Marina V. Rodnina: Max Planck Institute for Multidisciplinary Sciences
Sarah Adio: Institute for Microbiology and Genetics, Georg-August University of Göttingen

Nature Communications, 2022, vol. 13, issue 1, 1-15

Abstract: Abstract When reading consecutive mRNA codons, ribosomes move by exactly one triplet at a time to synthesize a correct protein. Some mRNA tracks, called slippery sequences, are prone to ribosomal frameshifting, because the same tRNA can read both 0- and –1-frame codon. Using smFRET we show that during EF-G-catalyzed translocation on slippery sequences a fraction of ribosomes spontaneously switches from rapid, accurate translation to a slow, frameshifting-prone translocation mode where the movements of peptidyl- and deacylated tRNA become uncoupled. While deacylated tRNA translocates rapidly, pept-tRNA continues to fluctuate between chimeric and posttranslocation states, which slows down the re-locking of the small ribosomal subunit head domain. After rapid release of deacylated tRNA, pept-tRNA gains unconstrained access to the –1-frame triplet, resulting in slippage followed by recruitment of the –1-frame aa-tRNA into the A site. Our data show how altered choreography of tRNA and ribosome movements reduces the translation fidelity of ribosomes translocating in a slow mode.

Date: 2022
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DOI: 10.1038/s41467-022-31852-w

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