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Related Concept Videos

Improving Translational Accuracy02:07

Improving Translational Accuracy

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Base complementarity between the three base pairs of mRNA codon and the tRNA anticodon is not a failsafe mechanism. Inaccuracies can range from a single mismatch to no correct base pairing at all. The free energy difference between the correct and nearly correct base pairs can be as small as 3 kcal/ mol. With complementarity being the only proofreading step, the estimated error frequency would be one wrong amino acid in every 100 amino acids incorporated. However, error frequencies observed in...
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Translational Regulation01:29

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Translational regulation in prokaryotes ensures efficient protein synthesis by controlling ribosome access to mRNA. This regulation is mediated by secondary RNA structures, including translational riboswitches, RNA thermometers, and small RNAs (sRNAs), which respond to intracellular and environmental signals to modulate gene expression.Translational RiboswitchesRiboswitches in the leader region of mRNAs can regulate translation by altering the accessibility of the Shine-Dalgarno (SD) sequence,...
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Termination of Translation01:44

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The large ribosomal subunit has several important structures essential to translation. These include the peptidyl transferase center (PTC) - which is the site where the peptide bond is formed - and a large, internal, water-filled tube through which the nascent polypeptide moves. This latter structure is called the Peptide Exit Tunnel, and it begins at the PTC and spans the body of the large ribosomal subunit. During translation, as the nascent polypeptide chain is synthesized, it passes through...
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Ribosome synthesis is a highly complex and coordinated process involving more than 200 assembly factors. The synthesis and processing of ribosomal components occurs not only in the nucleolus but also in the nucleoplasm and the cytoplasm of eukaryotic cells.
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Initiating translation is complex because it involves multiple molecules. Initiator tRNA, ribosomal subunits, and eukaryotic initiation factors (eIFs) are all required to assemble on the initiation codon of mRNA. This process consists of several steps that are mediated by different eIFs.
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Genome-wide Quantification of Translation in Budding Yeast by Ribosome Profiling
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Structural adaptations for enhanced translation kinetics in evolved ribosomes.

Tushar Raskar1,2, Alan Costello3,4, Ahmed H Badran3,4

  • 1Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, San Francisco, CA 94158.

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Summary

Engineered ribosomes with faster translation rates show increased protein yield. Subtle RNA sequence changes, not just catalytic centers, influence ribosome efficiency and structure.

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Area of Science:

  • Molecular Biology
  • Structural Biology
  • Biochemistry

Background:

  • Ribosomal RNA (rRNA) sequence dictates translation dynamics, but non-catalytic center modifications' effects on kinetics and protein yield are unclear.
  • Previous work utilized orthogonal ribosome phage-assisted continuous evolution (oRibo-PACE) to create chimeric ribosomes with enhanced translation rates.

Purpose of the Study:

  • To structurally characterize kinetically enhanced chimeric ribosomes.
  • To investigate the relationship between 16S rRNA stability and translation efficiency.
  • To establish principles for engineering ribosome translation properties.

Main Methods:

  • Cryo-electron microscopy (cryo-EM) for structural characterization.
  • Orthogonal ribosome phage-assisted continuous evolution (oRibo-PACE) for ribosome engineering.
  • Analysis of RNA structural adaptations and RNA-protein interactions.

Main Results:

  • Evolved ribosomes exhibit extensive 16S rRNA structural adaptations, including mismatches at helical junctions, leading to RNA-protein rearrangements.
  • Destabilization of non-canonical base pairs and local structural flexibility correlate with increased translational output.
  • Compensatory mutations restoring stability reduced translational activity to wild-type levels.

Conclusions:

  • 16S rRNA sequence malleability allows for significant structural adaptation influencing translation efficiency.
  • Subtle, localized destabilization of specific rRNA elements can enhance translational output.
  • Provides a framework for engineering ribosomes with tailored translation kinetics and protein yields.