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Related Concept Videos

Ribosomal RNA Synthesis02:53

Ribosomal RNA Synthesis

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.
Ribosome biogenesis begins with the synthesis of 5S and 45S pre-rRNAs by distinct RNA polymerases. The primary transcripts are extensively processed and modified before they are bound and folded by ribosomal proteins and assembly factors,...
Improving Translational Accuracy02:07

Improving Translational Accuracy

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...
Initiation of Translation02:33

Initiation of Translation

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.
First, the initiator tRNA must be selected from the pool of elongator tRNAs by eukaryotic initiation factor 2 (eIF2). The initiator tRNA (Met-tRNAi) has conserved sequence elements including modified bases at...
Leaky Scanning02:28

Leaky Scanning

During most eukaryotic translation processes, the small 40S ribosome subunit scans an mRNA from its 5' end until it encounters the first start AUG codon. The large 60S ribosomal subunit then joins the smaller one to initiate protein synthesis. The location of the translation initiation is largely determined by the nucleotides near the start codon as there may be multiple translation initiation sites present on the mRNA.  Marilyn Kozak discovered that the sequence RCCAUGG (where R stands for...
Ribosomal RNA Synthesis02:53

Ribosomal RNA Synthesis

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.
Ribosome biogenesis begins with the synthesis of 5S and 45S pre-rRNAs by distinct RNA polymerases. The primary transcripts are extensively processed and modified before they are bound and folded by ribosomal proteins and assembly factors,...
Initiation of Translation02:33

Initiation of Translation

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.
First, the initiator tRNA must be selected from the pool of elongator tRNAs by eukaryotic initiation factor 2 (eIF2). The initiator tRNA (Met-tRNAi) has conserved sequence elements including modified bases at...

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Related Experiment Video

Updated: Jul 25, 2026

Isolation of Ribosome Bound Nascent Polypeptides in vitro to Identify Translational Pause Sites Along mRNA
10:15

Isolation of Ribosome Bound Nascent Polypeptides in vitro to Identify Translational Pause Sites Along mRNA

Published on: July 6, 2012

Kinetics of ribosomal pausing during programmed -1 translational frameshifting.

J D Lopinski1, J D Dinman, J A Bruenn

  • 1Department of Biological Sciences, State University of New York at Buffalo, Buffalo, New York 14260, USA.

Molecular and Cellular Biology
|January 29, 2000
PubMed
Summary

Programmed ribosomal frameshifting in yeast viruses requires a pause site. Only one of three ribosome passage methods at this site results in frameshifting, influencing viral RNA translation.

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Last Updated: Jul 25, 2026

Isolation of Ribosome Bound Nascent Polypeptides in vitro to Identify Translational Pause Sites Along mRNA
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Published on: July 6, 2012

Optical Tweezers to Study RNA-Protein Interactions in Translation Regulation
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Single Molecule Fluorescence Energy Transfer Study of Ribosome Protein Synthesis
08:07

Single Molecule Fluorescence Energy Transfer Study of Ribosome Protein Synthesis

Published on: July 6, 2021

Area of Science:

  • Molecular Biology
  • Virology
  • Biochemistry

Background:

  • Programmed -1 ribosomal frameshifting is crucial for translating essential viral RNA in Saccharomyces cerevisiae.
  • This process relies on a specific slippery site and a downstream pseudoknot.
  • Previous studies indicated ribosomes pause at the slippery site during frameshifting.

Purpose of the Study:

  • To investigate the kinetics of ribosomal pausing and frameshifting at the slippery site.
  • To quantify the rates of synthesis, pause times, and frameshifting efficiency.
  • To elucidate the mechanisms by which ribosomes navigate the frameshift site.

Main Methods:

  • In vitro translation assays were employed.
  • Quantitative heelprinting was used to detect translational intermediates.
  • Ribosomal pause times and frameshifting fractions were estimated.

Main Results:

  • Approximately 10% of ribosomes paused at the slippery site in vitro.
  • About 60% of paused ribosomes proceeded in the -1 frame.
  • Ribosomes frameshifting into the -1 frame exhibited pause times significantly longer than peptide bond synthesis.
  • Altering translational initiation rates affected frameshifting in vivo.

Conclusions:

  • Three distinct mechanisms govern ribosome passage at the frameshift site.
  • Only one of these mechanisms leads to successful -1 ribosomal frameshifting.
  • The rate of translational initiation influences frameshifting efficiency in vivo.