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Related Concept Videos

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

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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,...
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Nonsense-mediated mRNA Decay02:27

Nonsense-mediated mRNA Decay

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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,...
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Signal Sequences and Sorting Receptors01:41

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Signal sequences are short amino acid sequences that guide newly synthesized proteins to their proper location within the cell. Classical signal sequences are fifteen to sixty amino acids long and present at the N-terminus of a polypeptide chain. Each signal sequence has a conserved segment of basic residues towards their N terminus, a hydrophobic core, and a C-terminus rich in polar residues. The C-terminus also contains a signal cleavage site and features a -3 -1 sequence motif. The -3-1...
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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...
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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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Signal and noise in circRNA translation.

T B Hansen1

  • 1MBG, Aarhus University, C.F. Moellers Alle 3, 8000 Aarhus C, Denmark.

Methods (San Diego, Calif.)
|February 15, 2021
PubMed
Summary

Circular RNAs (circRNAs) are increasingly studied for their roles in disease. Recent findings suggest circRNAs may produce proteins, but this remains debated and requires careful experimental validation.

Area of Science:

  • RNA biology
  • Molecular biology
  • Genetics

Background:

  • Circular RNAs (circRNAs) are a novel class of RNA molecules.
  • Initially considered non-coding, recent studies suggest circRNAs may be translated into proteins.
  • This potential protein-coding capacity of circRNAs is a subject of ongoing scientific debate.

Purpose of the Study:

  • To outline experimental methods for detecting circRNA translation.
  • To provide guidelines for distinguishing true signals from experimental noise in circRNA translation studies.
  • To address current controversies regarding circRNA protein production.

Main Methods:

  • Review of conventional experimental techniques for circRNA translation detection.
  • Discussion of data analysis strategies to validate findings.
Keywords:
ArtefactsCircular RNAMass spectrometryMass-specRiboSeqTranslationcircRNA

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  • Emphasis on rigorous experimental design.

    • Main Results:

    • Established methods for detecting circRNA translation are presented.
    • Guidelines are provided to ensure the reliability of experimental results.
    • The importance of distinguishing true translation signals from background noise is highlighted.

    Conclusions:

    • Accurate detection of circRNA translation requires careful methodology and data interpretation.
    • The presented guidelines are crucial for resolving current debates in the field.
    • These principles can also inform research on other forms of non-canonical translation.