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Related Concept Videos

Initiation of Translation02:33

Initiation of Translation

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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.
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

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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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PI3K/mTOR/AKT Signaling Pathway01:22

PI3K/mTOR/AKT Signaling Pathway

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The mammalian target of rapamycin  (mTOR) is a serine/threonine kinase that regulates growth, proliferation, and cell survival in response to hormones, growth factors, or nutrient availability. This kinase exists in two structurally and functionally distinct forms: mTOR complex 1  (mTORC1) and mTOR complex 2  (mTORC2). The first form (mTORC1) is composed of a rapamycin-sensitive Raptor and proline-rich Akt substrate, PRAS40. In contrast,  mTORC2 consists of a...
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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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Directing Proteins to the Rough Endoplasmic Reticulum01:34

Directing Proteins to the Rough Endoplasmic Reticulum

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The organelle-specific signaling sequences direct proteins synthesized in the cytosol to their final destination like ER, mitochondria, peroxisomes, etc. Some of the proteins directed to ER are then trafficked via vesicles to other organelles within the cell or the extracellular environment through the Golgi complex. For example, the rough ER synthesizes soluble proteins for transportation to the lysosomes or secretion out of the cell. It can also synthesize transmembrane proteins that can...
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Regulation of Expression Occurs at Multiple Steps02:24

Regulation of Expression Occurs at Multiple Steps

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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.
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Related Experiment Video

Updated: Jul 2, 2025

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

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miR-483-5p orchestrates the initiation of protein synthesis by facilitating the decrease in phosphorylated

Siranjeevi Nagaraj1,2, Anna Stankiewicz-Drogon3, Edward Darzynkiewicz1,3

  • 1Interdisciplinary Laboratory of Molecular Biology and Biophysics, Centre of New Technologies, University of Warsaw, 02-097, Warsaw, Poland.

Scientific Reports
|February 20, 2024
PubMed
Summary

MicroRNA-483-5p targets multiple proteins involved in protein synthesis, reducing phosphorylated eukaryotic initiation factor 4E (pSer209eIF4E) levels. This multi-targeting capability offers potential therapeutic strategies for diseases like cancer.

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Area of Science:

  • Molecular Biology
  • Gene Regulation
  • Cancer Research

Background:

  • Eukaryotic initiation factor 4E (eIF4E) is crucial for protein synthesis regulation in health and disease.
  • MicroRNAs (miRNAs) regulate gene expression by targeting mRNAs, but their role in eIF4E regulation is unclear.
  • Understanding miRNA-mediated regulation of eIF4E is vital for developing novel therapeutic approaches.

Purpose of the Study:

  • To investigate the regulatory role of miR-483-5p on eukaryotic translation initiation factor 4E (eIF4E) and its associated proteins.
  • To identify and experimentally verify the mRNA targets of miR-483-5p involved in protein synthesis.
  • To elucidate the effect of miR-483-5p on eIF4E phosphorylation.

Main Methods:

  • Bioinformatic prediction of miR-483-5p targets using TargetScan and Web of Science databases.
  • Experimental verification of miR-483-5p targets (ERK1, MKNK1, EIF4EBP1, EIF4EBP2, EIF4E) in HEK293 cells.
  • Quantitative analysis of mRNA levels and protein phosphorylation (pSer209eIF4E).

Main Results:

  • miR-483-5p was predicted to target EIF4E, EIF4EBP1, and EIF4EBP2.
  • miR-483-5p significantly reduced ERK1 and MKNK1 mRNA levels and suppressed EIF4EBP1 and EIF4EBP2 expression.
  • miR-483-5p decreased the phosphorylation of eIF4E at Ser209 (pSer209eIF4E) without affecting total eIF4E levels.

Conclusions:

  • miR-483-5p modulates eIF4E phosphorylation through a multi-targeting mechanism affecting the ERK1/MKNK1 pathway.
  • Unlike single-target siRNAs, miRNAs can regulate entire pathways, offering a distinct therapeutic advantage.
  • Further exploration of miR-483-5p in cancer models may reveal novel therapeutic opportunities.