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Related Concept Videos

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...
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...
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...
Translation in Prokaryotes01:29

Translation in Prokaryotes

Prokaryote translation is a complex, highly coordinated process that converts genetic information from mRNA into functional proteins. It involves three stages: initiation, elongation, and termination, each facilitated by specific molecular components.Initiation of TranslationThe process begins with the assembly of the ribosomal subunits and initiation factors on the mRNA. In bacteria, the 30S ribosomal subunit recognizes the Shine-Dalgarno sequence in the mRNA, a conserved region upstream of...
Ribosome Profiling02:24

Ribosome Profiling

Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique helps...
Protein Dynamics in Living Cells01:19

Protein Dynamics in Living Cells

Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...

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Related Experiment Video

Updated: Jun 6, 2026

Quantitative Immunofluorescence to Measure Global Localized Translation
09:13

Quantitative Immunofluorescence to Measure Global Localized Translation

Published on: August 22, 2017

Elucidating mechanistic principles underpinning eukaryotic translation initiation using quantitative fluorescence

Abigail L Stevenson1, Pedro P Juanes, John E G McCarthy

  • 1Manchester Interdisciplinary Biocentre, University of Manchester, 131 Princes Street, Manchester M1 7DN, UK.

Biochemical Society Transactions
|December 2, 2010
PubMed
Summary

Quantitative fluorescence methods like FRET and FA are advancing the study of eukaryotic translation initiation, a complex cellular process involving large molecular machines and multiple initiation factors (eIFs). These techniques offer new insights into intricate assembly pathways.

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Last Updated: Jun 6, 2026

Quantitative Immunofluorescence to Measure Global Localized Translation
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An In Vitro Single-Molecule Imaging Assay for the Analysis of Cap-Dependent Translation Kinetics
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Toeprinting Analysis of Translation Initiation Complex Formation on Mammalian mRNAs
10:37

Toeprinting Analysis of Translation Initiation Complex Formation on Mammalian mRNAs

Published on: May 10, 2018

Area of Science:

  • Molecular biology
  • Biochemistry
  • Cell biology

Background:

  • Eukaryotic translation initiation is a complex process involving over 11 eukaryotic initiation factors (eIFs) and ribosomes, forming large molecular machines.
  • Traditional molecular and structural methods face challenges in studying these large complexes.
  • Novel quantitative techniques are increasingly employed to analyze complex assembly pathways.

Purpose of the Study:

  • To introduce quantitative fluorescence methods for studying eukaryotic translation initiation.
  • To discuss the impact of these methods on understanding complex molecular assemblies.
  • To highlight recent methodological developments enhancing technique applicability.

Main Methods:

  • Labeling ribosomal subunits and/or eIFs with fluorophores.
  • Utilizing techniques such as Förster resonance energy transfer (FRET).
  • Employing fluorescence anisotropy (FA) measurements.

Main Results:

  • Quantitative fluorescence methods provide new ways to dissect complex assembly pathways.
  • Recent methodological advancements have improved the applicability of FRET and FA.
  • These techniques are beginning to significantly impact the field of translation initiation research.

Conclusions:

  • Quantitative fluorescence methods, including FRET and FA, are powerful tools for studying large macromolecular complexes like the translation initiation machinery.
  • These techniques offer enhanced insights into the dynamics and assembly of eukaryotic translation initiation.
  • Continued development and application of these methods will further advance our understanding of this fundamental cellular process.