Distinct XPPX sequence motifs induce ribosome stalling, which is rescued by the translation elongation factor EF-P
(2013) In Proceedings of the National Academy of Sciences of the United States of America 110(38). p.15265-15270- Abstract
Ribosomes are the protein synthesizing factories of the cell, polymerizing polypeptide chains from their constituent amino acids. However, distinct combinations of amino acids, such as polyproline stretches, cannot be efficiently polymerized by ribosomes, leading to translational stalling. The stalled ribosomes are rescued by the translational elongation factor P (EF-P), which by stimulating peptide-bond formation allows translation to resume. Using metabolic stable isotope labeling and mass spectrometry, we demonstrate in vivo that EF-P is important for expression of not only polyproline-containing proteins, but also for specific subsets of proteins containing diprolyl motifs (XPP/PPX). Together with a systematic in vitro and in vivo... (More)
Ribosomes are the protein synthesizing factories of the cell, polymerizing polypeptide chains from their constituent amino acids. However, distinct combinations of amino acids, such as polyproline stretches, cannot be efficiently polymerized by ribosomes, leading to translational stalling. The stalled ribosomes are rescued by the translational elongation factor P (EF-P), which by stimulating peptide-bond formation allows translation to resume. Using metabolic stable isotope labeling and mass spectrometry, we demonstrate in vivo that EF-P is important for expression of not only polyproline-containing proteins, but also for specific subsets of proteins containing diprolyl motifs (XPP/PPX). Together with a systematic in vitro and in vivo analysis, we provide a distinct hierarchy of stalling triplets, ranging from strong stallers, such as PPP, DPP, and PPN to weak stallers, such as CPP, PPR, and PPH, all of which are substrates for EF-P. These findings provide mechanistic insight into how the characteristics of the specific amino acid substrates influence the fundamentals of peptide bond formation.
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- author
- Peil, Lauri ; Starosta, Agata L ; Lassak, Jürgen ; Atkinson, Gemma C LU ; Virumäe, Kai ; Spitzer, Michaela ; Tenson, Tanel ; Jung, Kirsten ; Remme, Jaanus and Wilson, Daniel N
- publishing date
- 2013
- type
- Contribution to journal
- publication status
- published
- keywords
- Amino Acid Motifs/genetics, Chromatography, Liquid, Escherichia coli K12/metabolism, Humans, Peptide Elongation Factors/metabolism, Proline/metabolism, Protein Biosynthesis/physiology, Proteomics, Ribosomes/metabolism, Tandem Mass Spectrometry, beta-Galactosidase
- in
- Proceedings of the National Academy of Sciences of the United States of America
- volume
- 110
- issue
- 38
- pages
- 15265 - 15270
- publisher
- National Academy of Sciences
- external identifiers
-
- scopus:84884335094
- pmid:24003132
- ISSN
- 1091-6490
- DOI
- 10.1073/pnas.1310642110
- language
- English
- LU publication?
- no
- id
- 20ef5131-8719-48d8-b439-78aff6c87c54
- date added to LUP
- 2021-09-27 15:53:51
- date last changed
- 2024-08-10 22:07:38
@article{20ef5131-8719-48d8-b439-78aff6c87c54, abstract = {{<p>Ribosomes are the protein synthesizing factories of the cell, polymerizing polypeptide chains from their constituent amino acids. However, distinct combinations of amino acids, such as polyproline stretches, cannot be efficiently polymerized by ribosomes, leading to translational stalling. The stalled ribosomes are rescued by the translational elongation factor P (EF-P), which by stimulating peptide-bond formation allows translation to resume. Using metabolic stable isotope labeling and mass spectrometry, we demonstrate in vivo that EF-P is important for expression of not only polyproline-containing proteins, but also for specific subsets of proteins containing diprolyl motifs (XPP/PPX). Together with a systematic in vitro and in vivo analysis, we provide a distinct hierarchy of stalling triplets, ranging from strong stallers, such as PPP, DPP, and PPN to weak stallers, such as CPP, PPR, and PPH, all of which are substrates for EF-P. These findings provide mechanistic insight into how the characteristics of the specific amino acid substrates influence the fundamentals of peptide bond formation. </p>}}, author = {{Peil, Lauri and Starosta, Agata L and Lassak, Jürgen and Atkinson, Gemma C and Virumäe, Kai and Spitzer, Michaela and Tenson, Tanel and Jung, Kirsten and Remme, Jaanus and Wilson, Daniel N}}, issn = {{1091-6490}}, keywords = {{Amino Acid Motifs/genetics; Chromatography, Liquid; Escherichia coli K12/metabolism; Humans; Peptide Elongation Factors/metabolism; Proline/metabolism; Protein Biosynthesis/physiology; Proteomics; Ribosomes/metabolism; Tandem Mass Spectrometry; beta-Galactosidase}}, language = {{eng}}, number = {{38}}, pages = {{15265--15270}}, publisher = {{National Academy of Sciences}}, series = {{Proceedings of the National Academy of Sciences of the United States of America}}, title = {{Distinct XPPX sequence motifs induce ribosome stalling, which is rescued by the translation elongation factor EF-P}}, url = {{http://dx.doi.org/10.1073/pnas.1310642110}}, doi = {{10.1073/pnas.1310642110}}, volume = {{110}}, year = {{2013}}, }