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Related Concept Videos

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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The structure and stability of mRNA molecules regulates gene expression, as mRNAs are a key step in the pathway from gene to protein. In eukaryotes, the half-life of mRNA varies from a few minutes up to several days. mRNA stability is essential in growth and development. The absence of the proteins regulating its stability, such as tristetraprolin in mice, can cause systemic issues, including bone marrow overgrowth, inflammation, and autoimmunity.
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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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In eukaryotic cells, transcripts made by RNA polymerase are modified and processed before exiting the nucleus. Unprocessed RNA is called precursor mRNA or pre-mRNA to distinguish it from mature mRNA.
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Modulation of translational decoding by m6A modification of mRNA.

Sakshi Jain1, Lukasz Koziej2, Panagiotis Poulis1

  • 1Max Planck Institute for Multidisciplinary Sciences, Göttingen, 37077, Germany.

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N6-methyladenosine (m6A) modifies mRNA, impacting bacterial translation. This m6A modification hinders ribosome decoding by destabilizing complexes and increasing tRNA drop-off, affecting protein synthesis.

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Area of Science:

  • Molecular Biology
  • Biochemistry
  • Genetics

Background:

  • N6-methyladenosine (m6A) is a prevalent mRNA modification influencing mRNA metabolism.
  • m6A within mRNA coding regions is known to inhibit translation elongation.

Purpose of the Study:

  • To elucidate the mechanism by which m6A modulates decoding in bacterial translation systems.
  • To investigate the impact of m6A on ribosome-mRNA-tRNA interactions during translation.

Main Methods:

  • Utilized a combination of rapid kinetics, single-molecule Förster Resonance Energy Transfer (smFRET), and single-particle cryo-electron microscopy (cryo-EM).
  • Analyzed the stability of ribosome complexes and tRNA drop-off rates in the presence and absence of m6A.
  • Examined mRNA codon conformation and its remodeling upon aminoacyl-tRNA binding.

Main Results:

  • m6A does not impede the initial binding of aminoacyl-tRNA to the ribosome.
  • Fewer ribosomes successfully complete decoding in the presence of m6A due to reduced complex stability and increased tRNA drop-off.
  • m6A favors dynamic mRNA codon conformations that are less compatible with canonical codon-anticodon pairing and are rejected by the ribosome.

Conclusions:

  • m6A modifications, even outside the direct codon-anticodon interaction site, can interfere with base pairing and ribosome recognition.
  • These interactions modulate translational efficiency by affecting decoding fidelity and complex stability.
  • The study reveals a novel mechanism by which mRNA modifications regulate gene expression at the translational level.