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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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Related Experiment Video

Updated: Oct 11, 2025

Mass Spectrometry-Based Proteomics Analyses Using the OpenProt Database to Unveil Novel Proteins Translated from Non-Canonical Open Reading Frames
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The dark proteome: translation from noncanonical open reading frames.

Bradley W Wright1, Zixin Yi1, Jonathan S Weissman2

  • 1Laboratory of Functional Genomics and Translational Control, Cecil H. and Ida Green Center for Reproductive Biology Sciences, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA; Department of Pharmacology, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA; Harold C. Simmons Comprehensive Cancer Center, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA.

Trends in Cell Biology
|November 30, 2021
PubMed
Summary

Omics studies reveal many unannotated open reading frames (ORFs) with potential functions. This review discusses novel microproteins and regulatory roles of these noncanonical ORFs in mammals.

Keywords:
CRISPRmicroproteinsnoncanonical ORFsribosome profilingshort ORFstranslation

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Area of Science:

  • Genomics
  • Molecular Biology
  • Bioinformatics

Background:

  • Omics technologies have expanded the understanding of genomic coding potential.
  • Widespread unannotated open reading frames (ORFs) are prevalent across genomes.
  • These noncanonical ORFs may encode novel functional microproteins or regulatory elements.

Purpose of the Study:

  • To discuss recent advances in understanding noncanonical ORFs.
  • To explore the diversity and roles of these genomic regions.
  • To highlight biologically significant examples in the mammalian genome.

Main Methods:

  • Literature review of omics-based studies.
  • Analysis of genomic data for noncanonical ORF identification.
  • Synthesis of functional characterization data.

Main Results:

  • Noncanonical ORFs represent a significant, under-recognized component of the genome.
  • Functionally characterized noncanonical ORFs play essential roles in diverse biological processes.
  • Evidence suggests these regions encode novel microproteins and regulatory elements.

Conclusions:

  • Noncanonical ORFs are crucial for understanding the full coding potential of genomes.
  • Further research into these regions is essential for discovering novel biological functions.
  • Mammalian genomes harbor important examples of noncanonical ORF diversity and function.