Related Experiment Video
Updated: Feb 12, 2026
12:57
Genome-wide Quantification of Translation in Budding Yeast by Ribosome Profiling
Published on: December 21, 2017
12.0K
Genome-wide Translation Profiling by Ribosome-Bound tRNA Capture
Chien-Wen Chen1, Motomasa Tanaka1
1Laboratory for Protein Conformation Diseases, RIKEN Brain Science Institute, Hirosawa 2-1, Wako, Saitama 351-0198, Japan.
Cell Reports
|April 12, 2018
Summary
We developed a new method to profile transfer RNAs (tRNAs) within ribosomes during active translation. This technique reveals how environmental stress impacts tRNA quality, affecting protein synthesis and causing translation arrest.
Area of Science:
- Molecular Biology
- Cell Biology
- Biochemistry
Background:
- Transfer RNA (tRNA) plays a crucial role in messenger RNA (mRNA) translation within the ribosome.
- Cellular regulation during specific tRNA recruitment in polypeptide formation is hypothesized but poorly understood.
- Lack of technology to profile intra-ribosomal tRNAs has limited research in nascent chain biology.
Purpose of the Study:
- To characterize a novel technique for profiling ribosome-embedded tRNAs and their modifications.
- To generate a comprehensive survey of the quantity and quality of intra-ribosomal tRNAs.
- To investigate the role of tRNAs in cellular regulation and translation control.
Main Methods:
- Development and characterization of a tRNA ribosome profiling technique.
- Quantitative and qualitative analysis of intra-ribosomal tRNAs.
- Concurrent tRNA/mRNA ribosome profiling under environmental stress conditions.
Main Results:
- A comprehensive survey of intra-ribosomal tRNA quantity and quality was generated.
- Methionine tRNA within ribosomes serves as a biomarker for impaired translation initiation or elongation under stress.
- Stress-induced incorporation of damaged and uncharged tRNAs leads to translation arrest.
Conclusions:
- tRNA ribosome profiling offers new insights into translation control mechanisms.
- Environmental stress significantly impacts tRNA quality and its incorporation into ribosomes.
- The developed method can be applied to diverse biological contexts to study translation regulation.
Related Concept Videos
02:33Initiation of Translation
39.2K
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...
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...
39.2K
01:44Termination of Translation
27.8K
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...
27.8K
02:24Ribosome Profiling
4.2K
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...
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...
4.2K
01:31Translation
157.3K
Lesson: Translation
Translation is the process of synthesizing proteins from the genetic information carried by messenger RNA (mRNA). Following transcription, it constitutes the final step in the expression of genes. This process is carried out by ribosomes, complexes of protein and specialized RNA molecules. Ribosomes, transfer RNA (tRNA), and other proteins produce a chain of amino acids—the polypeptide—as the end product of translation.
Translation Produces the Building Blocks of...
Translation is the process of synthesizing proteins from the genetic information carried by messenger RNA (mRNA). Following transcription, it constitutes the final step in the expression of genes. This process is carried out by ribosomes, complexes of protein and specialized RNA molecules. Ribosomes, transfer RNA (tRNA), and other proteins produce a chain of amino acids—the polypeptide—as the end product of translation.
Translation Produces the Building Blocks of...
157.3K
02:07Improving Translational Accuracy
15.0K
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...
15.0K
01:27Ribosomes
77.2K
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 Assembly
Ribosomes 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...
Ribosome Structure and Assembly
Ribosomes 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...
77.2K