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Initiation of Translation02:33

Initiation of Translation

34.4K
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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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...
11.8K
Transcription Elongation Factors02:35

Transcription Elongation Factors

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Transcription elongation is a dynamic process that alters depending upon the sequence heterogeneity of the DNA being transcribed. Hence, it is not surprising that the elongation complex's composition also varies along the way while transcribing a gene.
The transcription elongation is regulated via pausing of RNA polymerase on several occasions during transcription. In bacteria, these halts are necessary because the transcription of DNA into mRNA is coupled to the translation of that mRNA...
11.1K
Leaky Scanning02:28

Leaky Scanning

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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...
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Translational Regulation01:29

Translational Regulation

88
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,...
88
Regulation of Expression at Multiple Steps01:23

Regulation of Expression at Multiple Steps

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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...
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Related Experiment Video

Updated: Sep 3, 2025

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

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mRNA- and factor-driven dynamic variability controls eIF4F-cap recognition for translation initiation.

Burak Çetin1, Seán E O'Leary1,2

  • 1Graduate Program in Cell, Molecular, and Developmental Biology, University of California Riverside, Riverside, CA 92521, USA.

Nucleic Acids Research
|July 24, 2022
PubMed
Summary
This summary is machine-generated.

Messenger RNA (mRNA) 5' cap recognition by eukaryotic initiation factor 4F (eIF4F) is crucial for translation. This study reveals how eIF4F subunit interactions with full-length mRNAs are kinetically controlled, influencing translation efficiency.

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Last Updated: Sep 3, 2025

Monitoring eIF4F Assembly by Measuring eIF4E-eIF4G Interaction in Live Cells
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Area of Science:

  • Molecular Biology
  • Biochemistry
  • Genetics

Background:

  • mRNA 5' cap recognition by the eukaryotic initiation factor 4F (eIF4F) complex is a critical regulatory step in protein translation.
  • Previous studies suggested kinetic differences in eIF4F-mRNA interactions contribute to translation efficiency variations, but detailed kinetic data for full-length mRNAs were lacking.

Purpose of the Study:

  • To investigate the real-time kinetics of eIF4F subunit interactions with full-length, polyadenylated mRNAs using single-molecule fluorescence.
  • To elucidate how mRNA length, eIF4G, eIF4A, and ATP influence these interactions and their role in translational control.

Main Methods:

  • Developed and utilized single-molecule fluorescence techniques to observe Saccharomyces cerevisiae eIF4F subunit dynamics with full-length mRNAs in real-time.
  • Analyzed association rates and the impact of mRNA length, eIF4G, eIF4A, and ATP hydrolysis on complex formation and dissociation.

Main Results:

  • eIF4E-mRNA association rates showed a linear inverse correlation with mRNA length.
  • eIF4G and eIF4A (with ATP) accelerated eIF4E-mRNA association, with effects modulated by mRNA length and cap-proximal secondary structure.
  • eIF4A-catalyzed ATP hydrolysis led to the ejection of eIF4E, suggesting a mechanism for ribosome recruitment preparation.

Conclusions:

  • mRNA-specific, factor-driven kinetics of eIF4F association play a significant role in regulating translation efficiency.
  • The findings provide a mechanistic insight into how eIF4F complex dynamics at the mRNA 5' end prepare for ribosome binding and initiate translation.