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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...
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...
Regulation of Expression at Multiple Steps01:23

Regulation of Expression at Multiple Steps

The gene expression in cells is regulated at different stages: (i) transcription, (ii) RNA processing, (iii) RNA localization, and (iv) translation. Transcriptional regulation is mediated by regulatory proteins such as transcription factors, activators, or repressors—these control gene expression by initiating or inhibiting the transcription of genes. Once a precursor or pre-mRNA is produced, it undergoes post-transcriptional modification, including 5' capping, splicing, and the addition of a...
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...
Regulation of Expression Occurs at Multiple Steps02:24

Regulation of Expression Occurs at Multiple Steps

Gene expression can be regulated at almost every step from gene to protein. Transcription is the step that is most commonly regulated. This involves the binding of proteins to short regulatory sequences on the DNA. This association can either promote or inhibit the transcription of a gene associated with the respective sequence.
Transcription results in the generation of precursor (pre-mRNA) that consists of both exons and introns, which needs further processing before being translated to a...

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Related Experiment Video

Updated: Jul 4, 2026

Monitoring eIF4F Assembly by Measuring eIF4E-eIF4G Interaction in Live Cells
08:47

Monitoring eIF4F Assembly by Measuring eIF4E-eIF4G Interaction in Live Cells

Published on: May 1, 2020

Dynamic redistribution of eIF4F controls cap-dependent translation initiation.

Riley C Gentry1,2, Nicholas A Ide1,3, Victoria M Comunale4

  • 1Department of Biological Sciences, Columbia University, New York, NY, USA.

Biorxiv : the Preprint Server for Biology
|July 3, 2026
PubMed
Summary

The eukaryotic initiation factor eIF4F binds mRNA caps to start translation. ATP binding aids eIF4F assembly, while ATP hydrolysis is crucial for recycling eIF4F during ribosome loading.

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Toeprinting Analysis of Translation Initiation Complex Formation on Mammalian mRNAs

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Related Experiment Videos

Last Updated: Jul 4, 2026

Monitoring eIF4F Assembly by Measuring eIF4E-eIF4G Interaction in Live Cells
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Published on: May 1, 2020

Analysis of Cap-binding Proteins in Human Cells Exposed to Physiological Oxygen Conditions
10:40

Analysis of Cap-binding Proteins in Human Cells Exposed to Physiological Oxygen Conditions

Published on: December 28, 2016

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:

  • Translation initiation is a critical step in gene expression regulated by eukaryotic initiation factor eIF4F.
  • eIF4F binds the 5' cap of messenger RNA (mRNA) to recruit the ribosomal pre-initiation complex (PIC).
  • The precise mechanism of eIF4F in stimulating PIC loading and the role of ATP hydrolysis remain unclear.

Purpose of the Study:

  • To elucidate the molecular mechanism of eIF4F during translation initiation.
  • To investigate the role of ATP binding and hydrolysis in eIF4F function.
  • To identify PIC-associated factors involved in eIF4F recycling.

Main Methods:

  • Single-molecule fluorescence microscopy was employed to directly observe eIF4F dynamics.
  • Experiments focused on cap recognition and PIC engagement stages of translation initiation.
  • The roles of ATP binding, hydrolysis, and specific initiation factors were assessed.

Main Results:

  • ATP binding, not hydrolysis, facilitates eIF4F assembly on mRNA and its redistribution.
  • ATP hydrolysis is essential for the recycling of eIF4F during productive PIC engagement.
  • eIF3 and eIF4B were identified as key PIC factors stimulating ATP-hydrolysis-dependent eIF4F recycling.

Conclusions:

  • A model is proposed where PIC engagement triggers ATP-hydrolysis-dependent eIF4F recycling, coupling eIF4F dynamics to PIC loading.
  • This mechanism explains how mRNA features and initiation factors regulate translational efficiency.
  • The findings provide a framework for understanding the control of translation initiation.