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Related Concept Videos

Ribosome Profiling02:24

Ribosome Profiling

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Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique...
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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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Termination of Translation01:44

Termination of Translation

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The large ribosomal subunit has several important structures essential to translation. These include the peptidyl transferase center (PTC) - which is the site where the peptide bond is formed - and a large, internal, water-filled tube through which the nascent polypeptide moves. This latter structure is called the Peptide Exit Tunnel, and it begins at the PTC and spans the body of the large ribosomal subunit. During translation, as the nascent polypeptide chain is synthesized, it passes through...
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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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Riboswitches01:56

Riboswitches

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Riboswitches are non-coding mRNA domains that regulate the transcription and translation of downstream genes without the help of proteins. Riboswitches bind directly to a metabolite and can form unique stem-loop or hairpin structures in response to the amount of the metabolite present. They have two distinct regions – a metabolite-binding aptamer and an expression platform.
The aptamer has high specificity for a particular metabolite which allows riboswitches to specifically regulate...
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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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Updated: Mar 19, 2026

Polysome Fractionation and Analysis of Mammalian Translatomes on a Genome-wide Scale
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Polysome Fractionation and Analysis of Mammalian Translatomes on a Genome-wide Scale

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Characterizing inactive ribosomes in translational profiling.

Botao Liu1, Shu-Bing Qian2

  • 1Division of Nutritional Sciences, Cornell University, Ithaca, NY, USA; Graduate Field of Genetics, Genomics, and Development, Cornell University, Ithaca, NY, USA; Present address: Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, MA, USA.

Translation (Austin, Tex.)
|June 24, 2016
PubMed
Summary
This summary is machine-generated.

Inactive ribosomes are found in the monosome fraction, offering a new way to study translation. This discovery provides insights into ribosome availability and translational profiling.

Keywords:
elongation factorsprofilingribosometranslation

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Global Identification of Co-Translational Interaction Networks by Selective Ribosome Profiling
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Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • Translational regulation is crucial for gene expression, with ribosomes existing as polysomes during active translation.
  • The role of monosomes in translational profiling has been historically underestimated.
  • Technological advancements enable genome-wide analysis of gene expression and translational control.

Purpose of the Study:

  • To investigate the composition and function of the monosome fraction in translational profiling.
  • To identify markers of ribosome activity and inactivity within different ribosome populations.
  • To develop a quantitative method for assessing ribosome availability.

Main Methods:

  • Isolation of monosome and polysome fractions using sucrose sedimentation.
  • Analysis of ribosome-protected footprints via ribosome profiling.
  • Detection of specific elongation factors (eEF2, eEF1A) associated with ribosomes.

Main Results:

  • The monosome fraction contains a significant population of inactive ribosomes not engaged in translation.
  • Elongation factor eEF2, but not eEF1A, is stably associated with these inactive monosomes.
  • Ribosome profiling of monosomes shows distinct but similar footprint patterns compared to polysomes.
  • Free ribosomal subunits contribute minimally to occupancy outside coding regions.

Conclusions:

  • The monosome fraction provides a valuable resource for studying ribosome status under various conditions.
  • The presence of eEF2 in inactive monosomes offers a unique marker for evaluating ribosome activity.
  • This study enhances translational profiling analysis by incorporating monosome data and revealing new layers of ribosome regulation.