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Related Concept Videos

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...
Nonsense-mediated mRNA Decay02:27

Nonsense-mediated mRNA Decay

The Upf proteins that carry out nonsense-mediated decay (NMD) are found in all eukaryotic organisms, including humans. Each protein has an individual role, but they need to work in collaboration. Upf1 is an ATP-dependent RNA helicase that unwinds the RNA helix. Because Upf1 can unwind any RNA, Upf2 and Upf3 are required to help Upf1 discriminate between nonsense and normal mRNAs.
Usually, Upf3 binds to an Exon Junction Complex (EJC) at mRNA splice sites. If a ribosome fully translates the mRNA,...
Nonsense-mediated mRNA Decay02:27

Nonsense-mediated mRNA Decay

The Upf proteins that carry out nonsense-mediated decay (NMD) are found in all eukaryotic organisms, including humans. Each protein has an individual role, but they need to work in collaboration. Upf1 is an ATP-dependent RNA helicase that unwinds the RNA helix. Because Upf1 can unwind any RNA, Upf2 and Upf3 are required to help Upf1 discriminate between nonsense and normal mRNAs.
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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...
Nuclear Export of mRNA02:31

Nuclear Export of mRNA

Before mRNAs are exported to the cytoplasm, it is crucial to check each mRNA for structural and functional integrity. Eukaryotic cells use several different mechanisms, collectively known as mRNA surveillance, to look for irregularities in mRNAs. Irregular or aberrant mRNA are rapidly degraded by various enzymes. If a defective mRNA escapes the surveillance, it would be translated into a protein which would either be non-functional or not function properly. One of the primary irregularities in...
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Inhibitors of Bacterial Protein Synthesis

Aminoglycosides constitute a highly potent class of bactericidal antibiotics that exert their antimicrobial effects by targeting the bacterial ribosome, specifically disrupting protein synthesis. These polycationic molecules consist of amino-modified sugars linked via glycosidic bonds to an aminocyclitol core such as 2-deoxystreptamine or streptamine. Their strong positive charges facilitate tight binding to the negatively charged phosphate backbone of ribosomal RNA (rRNA), primarily at the 16S...

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Related Experiment Video

Updated: Jun 5, 2026

Toeprinting Analysis of Translation Initiation Complex Formation on Mammalian mRNAs
10:37

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Published on: May 10, 2018

Identification of compounds that decrease the fidelity of start codon recognition by the eukaryotic translational

Julie E Takacs1, Timothy B Neary, Nicholas T Ingolia

  • 1Department of Biophysics and Biophysical Chemistry, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA.

RNA (New York, N.Y.)
|January 12, 2011
PubMed
Summary
This summary is machine-generated.

Researchers discovered two novel compounds that reduce the accuracy of start codon recognition during eukaryotic translation initiation. These small molecules provide new tools for studying translation and its regulation.

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Last Updated: Jun 5, 2026

Toeprinting Analysis of Translation Initiation Complex Formation on Mammalian mRNAs
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Published on: May 10, 2018

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Assessment of Selective mRNA Translation in Mammalian Cells by Polysome Profiling
10:00

Assessment of Selective mRNA Translation in Mammalian Cells by Polysome Profiling

Published on: October 28, 2014

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • Eukaryotic translation initiation involves numerous protein factors.
  • Defects in specific factors (e.g., eukaryotic initiation factor 1) can impair start codon fidelity, leading to the Sui(-) phenotype.
  • Small molecules that modulate start codon recognition fidelity are currently unknown.

Purpose of the Study:

  • To identify small molecules that decrease the fidelity of start codon recognition during translation initiation.
  • To develop chemical tools for investigating the molecular mechanisms of translation initiation and regulation.

Main Methods:

  • High-throughput screening of over 55,000 compounds using a dual luciferase assay in Saccharomyces cerevisiae.
  • Validation of compound effects using in vivo assays monitoring translation from non-AUG start codons.
  • Assessment of compound interactions with eukaryotic initiation factor 1 (eIF1) in yeast strains.

Main Results:

  • Two structurally related compounds were identified that reduce start codon selection fidelity approximately twofold.
  • These compounds enhance translation from non-AUG start codons, including a natural upstream open reading frame initiating at UUG.
  • The compounds exacerbate the Sui(-) phenotype caused by eIF1 haploinsufficiency or mutation.
  • Compound effects are suppressed by eIF1 overexpression, which restores fidelity in other Sui(-) contexts.

Conclusions:

  • The identified compounds directly impact the translational machinery, reducing the accuracy of AUG start codon selection.
  • These molecules serve as valuable chemical probes for dissecting the complex process of translation initiation.
  • The findings offer new avenues for understanding and potentially manipulating gene expression at the translational level.