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mRNA- and factor-driven dynamic variability controls eIF4F-cap recognition for translation initiation

mRNA 5′ cap recognition by eIF4F is a key element of eukaryotic translational control. Kinetic differences in eIF4F–mRNA interactions have long been proposed to mediate translation-efficiency differences between mRNAs, and recent transcriptome-wide studies have revealed significant heterogeneity in...

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Autores principales: Çetin, Burak, O’Leary, Seán E
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9371892/
https://www.ncbi.nlm.nih.gov/pubmed/35871304
http://dx.doi.org/10.1093/nar/gkac631
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author Çetin, Burak
O’Leary, Seán E
author_facet Çetin, Burak
O’Leary, Seán E
author_sort Çetin, Burak
collection PubMed
description mRNA 5′ cap recognition by eIF4F is a key element of eukaryotic translational control. Kinetic differences in eIF4F–mRNA interactions have long been proposed to mediate translation-efficiency differences between mRNAs, and recent transcriptome-wide studies have revealed significant heterogeneity in eIF4F engagement with differentially-translated mRNAs. However, detailed kinetic information exists only for eIF4F interactions with short model RNAs. We developed and applied single-molecule fluorescence approaches to directly observe real-time Saccharomyces cerevisiae eIF4F subunit interactions with full-length polyadenylated mRNAs. We found that eIF4E–mRNA association rates linearly anticorrelate with mRNA length. eIF4G–mRNA interaction accelerates eIF4E–mRNA association in proportion to mRNA length, as does an eIF4F-independent activity of eIF4A, though cap-proximal secondary structure still plays an important role in defining the final association rates. eIF4F–mRNA interactions remained dominated by effects of eIF4G, but were modulated to different extents for different mRNAs by the presence of eIF4A and ATP. We also found that eIF4A-catalyzed ATP hydrolysis ejects eIF4E, and likely eIF4E•eIF4G from the mRNA after initial eIF4F•mRNA complex formation, suggesting a mechanism to prepare the mRNA 5′ end for ribosome recruitment. Our results support a role for mRNA-specific, factor-driven eIF4F association rates in kinetically controlling translation.
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spelling pubmed-93718922022-08-12 mRNA- and factor-driven dynamic variability controls eIF4F-cap recognition for translation initiation Çetin, Burak O’Leary, Seán E Nucleic Acids Res RNA and RNA-protein complexes mRNA 5′ cap recognition by eIF4F is a key element of eukaryotic translational control. Kinetic differences in eIF4F–mRNA interactions have long been proposed to mediate translation-efficiency differences between mRNAs, and recent transcriptome-wide studies have revealed significant heterogeneity in eIF4F engagement with differentially-translated mRNAs. However, detailed kinetic information exists only for eIF4F interactions with short model RNAs. We developed and applied single-molecule fluorescence approaches to directly observe real-time Saccharomyces cerevisiae eIF4F subunit interactions with full-length polyadenylated mRNAs. We found that eIF4E–mRNA association rates linearly anticorrelate with mRNA length. eIF4G–mRNA interaction accelerates eIF4E–mRNA association in proportion to mRNA length, as does an eIF4F-independent activity of eIF4A, though cap-proximal secondary structure still plays an important role in defining the final association rates. eIF4F–mRNA interactions remained dominated by effects of eIF4G, but were modulated to different extents for different mRNAs by the presence of eIF4A and ATP. We also found that eIF4A-catalyzed ATP hydrolysis ejects eIF4E, and likely eIF4E•eIF4G from the mRNA after initial eIF4F•mRNA complex formation, suggesting a mechanism to prepare the mRNA 5′ end for ribosome recruitment. Our results support a role for mRNA-specific, factor-driven eIF4F association rates in kinetically controlling translation. Oxford University Press 2022-07-25 /pmc/articles/PMC9371892/ /pubmed/35871304 http://dx.doi.org/10.1093/nar/gkac631 Text en © The Author(s) 2022. Published by Oxford University Press on behalf of Nucleic Acids Research. https://creativecommons.org/licenses/by-nc/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (https://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com
spellingShingle RNA and RNA-protein complexes
Çetin, Burak
O’Leary, Seán E
mRNA- and factor-driven dynamic variability controls eIF4F-cap recognition for translation initiation
title mRNA- and factor-driven dynamic variability controls eIF4F-cap recognition for translation initiation
title_full mRNA- and factor-driven dynamic variability controls eIF4F-cap recognition for translation initiation
title_fullStr mRNA- and factor-driven dynamic variability controls eIF4F-cap recognition for translation initiation
title_full_unstemmed mRNA- and factor-driven dynamic variability controls eIF4F-cap recognition for translation initiation
title_short mRNA- and factor-driven dynamic variability controls eIF4F-cap recognition for translation initiation
title_sort mrna- and factor-driven dynamic variability controls eif4f-cap recognition for translation initiation
topic RNA and RNA-protein complexes
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9371892/
https://www.ncbi.nlm.nih.gov/pubmed/35871304
http://dx.doi.org/10.1093/nar/gkac631
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