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Selective Translation Complex Profiling Reveals Staged Initiation and Co-translational Assembly of Initiation Factor Complexes

Translational control targeting the initiation phase is central to the regulation of gene expression. Understanding all of its aspects requires substantial technological advancements. Here we modified yeast translation complex profile sequencing (TCP-seq), related to ribosome profiling, and adapted...

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Autores principales: Wagner, Susan, Herrmannová, Anna, Hronová, Vladislava, Gunišová, Stanislava, Sen, Neelam D., Hannan, Ross D., Hinnebusch, Alan G., Shirokikh, Nikolay E., Preiss, Thomas, Valášek, Leoš Shivaya
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cell Press 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7447980/
https://www.ncbi.nlm.nih.gov/pubmed/32589964
http://dx.doi.org/10.1016/j.molcel.2020.06.004
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author Wagner, Susan
Herrmannová, Anna
Hronová, Vladislava
Gunišová, Stanislava
Sen, Neelam D.
Hannan, Ross D.
Hinnebusch, Alan G.
Shirokikh, Nikolay E.
Preiss, Thomas
Valášek, Leoš Shivaya
author_facet Wagner, Susan
Herrmannová, Anna
Hronová, Vladislava
Gunišová, Stanislava
Sen, Neelam D.
Hannan, Ross D.
Hinnebusch, Alan G.
Shirokikh, Nikolay E.
Preiss, Thomas
Valášek, Leoš Shivaya
author_sort Wagner, Susan
collection PubMed
description Translational control targeting the initiation phase is central to the regulation of gene expression. Understanding all of its aspects requires substantial technological advancements. Here we modified yeast translation complex profile sequencing (TCP-seq), related to ribosome profiling, and adapted it for mammalian cells. Human TCP-seq, capable of capturing footprints of 40S subunits (40Ss) in addition to 80S ribosomes (80Ss), revealed that mammalian and yeast 40Ss distribute similarly across 5′TRs, indicating considerable evolutionary conservation. We further developed yeast and human selective TCP-seq (Sel-TCP-seq), enabling selection of 40Ss and 80Ss associated with immuno-targeted factors. Sel-TCP-seq demonstrated that eIF2 and eIF3 travel along 5′ UTRs with scanning 40Ss to successively dissociate upon AUG recognition; notably, a proportion of eIF3 lingers on during the initial elongation cycles. Highlighting Sel-TCP-seq versatility, we also identified four initiating 48S conformational intermediates, provided novel insights into ATF4 and GCN4 mRNA translational control, and demonstrated co-translational assembly of initiation factor complexes.
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spelling pubmed-74479802020-08-31 Selective Translation Complex Profiling Reveals Staged Initiation and Co-translational Assembly of Initiation Factor Complexes Wagner, Susan Herrmannová, Anna Hronová, Vladislava Gunišová, Stanislava Sen, Neelam D. Hannan, Ross D. Hinnebusch, Alan G. Shirokikh, Nikolay E. Preiss, Thomas Valášek, Leoš Shivaya Mol Cell Article Translational control targeting the initiation phase is central to the regulation of gene expression. Understanding all of its aspects requires substantial technological advancements. Here we modified yeast translation complex profile sequencing (TCP-seq), related to ribosome profiling, and adapted it for mammalian cells. Human TCP-seq, capable of capturing footprints of 40S subunits (40Ss) in addition to 80S ribosomes (80Ss), revealed that mammalian and yeast 40Ss distribute similarly across 5′TRs, indicating considerable evolutionary conservation. We further developed yeast and human selective TCP-seq (Sel-TCP-seq), enabling selection of 40Ss and 80Ss associated with immuno-targeted factors. Sel-TCP-seq demonstrated that eIF2 and eIF3 travel along 5′ UTRs with scanning 40Ss to successively dissociate upon AUG recognition; notably, a proportion of eIF3 lingers on during the initial elongation cycles. Highlighting Sel-TCP-seq versatility, we also identified four initiating 48S conformational intermediates, provided novel insights into ATF4 and GCN4 mRNA translational control, and demonstrated co-translational assembly of initiation factor complexes. Cell Press 2020-08-20 /pmc/articles/PMC7447980/ /pubmed/32589964 http://dx.doi.org/10.1016/j.molcel.2020.06.004 Text en © 2020 The Author(s) http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Article
Wagner, Susan
Herrmannová, Anna
Hronová, Vladislava
Gunišová, Stanislava
Sen, Neelam D.
Hannan, Ross D.
Hinnebusch, Alan G.
Shirokikh, Nikolay E.
Preiss, Thomas
Valášek, Leoš Shivaya
Selective Translation Complex Profiling Reveals Staged Initiation and Co-translational Assembly of Initiation Factor Complexes
title Selective Translation Complex Profiling Reveals Staged Initiation and Co-translational Assembly of Initiation Factor Complexes
title_full Selective Translation Complex Profiling Reveals Staged Initiation and Co-translational Assembly of Initiation Factor Complexes
title_fullStr Selective Translation Complex Profiling Reveals Staged Initiation and Co-translational Assembly of Initiation Factor Complexes
title_full_unstemmed Selective Translation Complex Profiling Reveals Staged Initiation and Co-translational Assembly of Initiation Factor Complexes
title_short Selective Translation Complex Profiling Reveals Staged Initiation and Co-translational Assembly of Initiation Factor Complexes
title_sort selective translation complex profiling reveals staged initiation and co-translational assembly of initiation factor complexes
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7447980/
https://www.ncbi.nlm.nih.gov/pubmed/32589964
http://dx.doi.org/10.1016/j.molcel.2020.06.004
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