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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.
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The technique helps...

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Ribosomes

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Ribosomes translate genetic information encoded by messenger RNA (mRNA) into proteins. Both prokaryotic and eukaryotic cells have ribosomes. Cells that synthesize large quantities of protein—such as secretory cells in the human pancreas—can contain millions of ribosomes.Ribosome Structure and AssemblyRibosomes are composed of ribosomal RNA (rRNA) and proteins. In eukaryotes, rRNA is transcribed from genes in the nucleolus—a part of the nucleus that specializes in ribosome production. Within the...

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Ribosomes

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Ribosomes translate genetic information encoded by messenger RNA (mRNA) into proteins. Both prokaryotic and eukaryotic cells have ribosomes. Cells that synthesize large quantities of protein—such as secretory cells in the human pancreas—can contain millions of ribosomes.Ribosome Structure and AssemblyRibosomes are composed of ribosomal RNA (rRNA) and proteins. In eukaryotes, rRNA is transcribed from genes in the nucleolus—a part of the nucleus that specializes in ribosome production. Within the...

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Updated: Jun 10, 2026

In Vitro Reassociation Assay to Measure the Formation of 80S Ribosomal Particles Using Salt-washed Ribosomal Subunits

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Observing intersubunit dynamics in single yeast ribosomes.

Ananya Das¹, Amy K Grove¹, Aleksandr V Ivanov¹

¹Department of Biochemistry & Biophysics, School of Medicine and Dentistry, and Center for RNA Biology, University of Rochester, Rochester, NY 14642,United States.

Nucleic Acids Research

|June 9, 2026

Summary

This summary is machine-generated.

Yeast ribosomes rotate between nonrotated and rotated states during translation. Elongation factor 3 (eEF3) stabilizes the nonrotated state, influencing tRNA release and translation regulation.

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Area of Science:

Molecular Biology
Biochemistry
Structural Biology

Background:

Ribosomes translate genetic information into proteins.
Ribosome subunit rotation is crucial for translation.
Eukaryotic translation involves complex dynamics and regulatory factors.

Purpose of the Study:

To investigate the intersubunit dynamics of Saccharomyces cerevisiae ribosomes during translation.
To elucidate the role of fungal elongation factor 3 (eEF3) in ribosome conformation.
To understand the coupling of ribosome rotation with tRNA binding states.

Main Methods:

Single-molecule Förster resonance energy transfer (smFRET) assay.
Utilized fluorophores on ribosomal proteins uS15 and eL30.
Monitored FRET states corresponding to nonrotated (NR) and rotated (R) conformations.

Main Results:

Yeast ribosomes sample two predominant FRET states (NR and R).
Intersubunit rotation is coupled to tRNA transitions between classical and hybrid binding states.
The elongation cycle involves switching between NR and R conformations.
eEF3 stabilizes the NR conformation, facilitating E-site tRNA release.

Conclusions:

eEF3 plays a key role in stabilizing the nonrotated ribosome conformation.
Ribosome dynamics, influenced by eEF3, are critical for efficient translation and regulation.
The uS15-eL30 smFRET assay is a valuable tool for studying eukaryotic translation mechanisms.