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Assessment of Selective mRNA Translation in Mammalian Cells by Polysome Profiling
Published on: October 28, 2014
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Protocol for quantifying silent ribosome induction in yeast and mammalian cells using polysome profiling.
Sayanur Rahaman1, Samuel Mondal1, Nicole Schiffelholz1
1Biozentrum, Universität Basel, Basel 4056, Switzerland.
STAR Protocols
|March 22, 2026
Summary
This study introduces a new protocol to quantify translationally silent ribosomes, which lack messenger RNA (mRNA) and are hard to measure. The method works in yeast and mammalian cells, enabling better understanding of ribosome regulation.
Area of Science:
- Molecular Biology
- Cell Biology
- Biochemistry
Background:
- Translationally silent ribosomes, which lack bound mRNA, are challenging to quantify.
- Understanding ribosome dynamics is crucial for cellular regulation and disease research.
Purpose of the Study:
- To present a robust protocol for measuring silent ribosome induction in both yeast and mammalian cells.
- To enable quantitative analysis of ribosome states under various experimental conditions.
Main Methods:
- Isolation of polysome-profiling fractions.
- RNA sequencing of ribosome-associated RNA, including identification and removal of anomalously amplified ribosomal RNAs (rRNAs).
- Validation of measurements using quantitative polymerase chain reaction (qPCR).
Main Results:
- The protocol successfully quantifies silent ribosome induction across diverse conditions.
- The method allows for accurate analysis of ribosome-associated RNA, distinguishing functional and silent ribosomes.
- rRNA amplification artifacts can be identified and corrected for reliable quantification.
Conclusions:
- This protocol provides a standardized approach to measure silent ribosomes, advancing the study of translational control.
- The developed method facilitates research into ribosome regulation in yeast and mammalian systems.
- Accurate quantification of silent ribosomes opens new avenues for investigating cellular responses and disease mechanisms.
