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Regulation of 3′ splice site selection after step 1 of splicing by spliceosomal C* proteins

Alternative precursor messenger RNA splicing is instrumental in expanding the proteome of higher eukaryotes, and changes in 3′ splice site (3'ss) usage contribute to human disease. We demonstrate by small interfering RNA–mediated knockdowns, followed by RNA sequencing, that many proteins first...

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Autores principales: Dybkov, Olexandr, Preußner, Marco, El Ayoubi, Leyla, Feng, Vivi-Yun, Harnisch, Caroline, Merz, Kilian, Leupold, Paula, Yudichev, Peter, Agafonov, Dmitry E., Will, Cindy L., Girard, Cyrille, Dienemann, Christian, Urlaub, Henning, Kastner, Berthold, Heyd, Florian, Lührmann, Reinhard
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Association for the Advancement of Science 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9984181/
https://www.ncbi.nlm.nih.gov/pubmed/36867703
http://dx.doi.org/10.1126/sciadv.adf1785
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author Dybkov, Olexandr
Preußner, Marco
El Ayoubi, Leyla
Feng, Vivi-Yun
Harnisch, Caroline
Merz, Kilian
Leupold, Paula
Yudichev, Peter
Agafonov, Dmitry E.
Will, Cindy L.
Girard, Cyrille
Dienemann, Christian
Urlaub, Henning
Kastner, Berthold
Heyd, Florian
Lührmann, Reinhard
author_facet Dybkov, Olexandr
Preußner, Marco
El Ayoubi, Leyla
Feng, Vivi-Yun
Harnisch, Caroline
Merz, Kilian
Leupold, Paula
Yudichev, Peter
Agafonov, Dmitry E.
Will, Cindy L.
Girard, Cyrille
Dienemann, Christian
Urlaub, Henning
Kastner, Berthold
Heyd, Florian
Lührmann, Reinhard
author_sort Dybkov, Olexandr
collection PubMed
description Alternative precursor messenger RNA splicing is instrumental in expanding the proteome of higher eukaryotes, and changes in 3′ splice site (3'ss) usage contribute to human disease. We demonstrate by small interfering RNA–mediated knockdowns, followed by RNA sequencing, that many proteins first recruited to human C* spliceosomes, which catalyze step 2 of splicing, regulate alternative splicing, including the selection of alternatively spliced NAGNAG 3′ss. Cryo–electron microscopy and protein cross-linking reveal the molecular architecture of these proteins in C* spliceosomes, providing mechanistic and structural insights into how they influence 3'ss usage. They further elucidate the path of the 3′ region of the intron, allowing a structure-based model for how the C* spliceosome potentially scans for the proximal 3′ss. By combining biochemical and structural approaches with genome-wide functional analyses, our studies reveal widespread regulation of alternative 3′ss usage after step 1 of splicing and the likely mechanisms whereby C* proteins influence NAGNAG 3′ss choices.
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spelling pubmed-99841812023-03-04 Regulation of 3′ splice site selection after step 1 of splicing by spliceosomal C* proteins Dybkov, Olexandr Preußner, Marco El Ayoubi, Leyla Feng, Vivi-Yun Harnisch, Caroline Merz, Kilian Leupold, Paula Yudichev, Peter Agafonov, Dmitry E. Will, Cindy L. Girard, Cyrille Dienemann, Christian Urlaub, Henning Kastner, Berthold Heyd, Florian Lührmann, Reinhard Sci Adv Biomedicine and Life Sciences Alternative precursor messenger RNA splicing is instrumental in expanding the proteome of higher eukaryotes, and changes in 3′ splice site (3'ss) usage contribute to human disease. We demonstrate by small interfering RNA–mediated knockdowns, followed by RNA sequencing, that many proteins first recruited to human C* spliceosomes, which catalyze step 2 of splicing, regulate alternative splicing, including the selection of alternatively spliced NAGNAG 3′ss. Cryo–electron microscopy and protein cross-linking reveal the molecular architecture of these proteins in C* spliceosomes, providing mechanistic and structural insights into how they influence 3'ss usage. They further elucidate the path of the 3′ region of the intron, allowing a structure-based model for how the C* spliceosome potentially scans for the proximal 3′ss. By combining biochemical and structural approaches with genome-wide functional analyses, our studies reveal widespread regulation of alternative 3′ss usage after step 1 of splicing and the likely mechanisms whereby C* proteins influence NAGNAG 3′ss choices. American Association for the Advancement of Science 2023-03-03 /pmc/articles/PMC9984181/ /pubmed/36867703 http://dx.doi.org/10.1126/sciadv.adf1785 Text en Copyright © 2023 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY-NC). 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 use, distribution, and reproduction in any medium, so long as the resultant use is not for commercial advantage and provided the original work is properly cited.
spellingShingle Biomedicine and Life Sciences
Dybkov, Olexandr
Preußner, Marco
El Ayoubi, Leyla
Feng, Vivi-Yun
Harnisch, Caroline
Merz, Kilian
Leupold, Paula
Yudichev, Peter
Agafonov, Dmitry E.
Will, Cindy L.
Girard, Cyrille
Dienemann, Christian
Urlaub, Henning
Kastner, Berthold
Heyd, Florian
Lührmann, Reinhard
Regulation of 3′ splice site selection after step 1 of splicing by spliceosomal C* proteins
title Regulation of 3′ splice site selection after step 1 of splicing by spliceosomal C* proteins
title_full Regulation of 3′ splice site selection after step 1 of splicing by spliceosomal C* proteins
title_fullStr Regulation of 3′ splice site selection after step 1 of splicing by spliceosomal C* proteins
title_full_unstemmed Regulation of 3′ splice site selection after step 1 of splicing by spliceosomal C* proteins
title_short Regulation of 3′ splice site selection after step 1 of splicing by spliceosomal C* proteins
title_sort regulation of 3′ splice site selection after step 1 of splicing by spliceosomal c* proteins
topic Biomedicine and Life Sciences
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9984181/
https://www.ncbi.nlm.nih.gov/pubmed/36867703
http://dx.doi.org/10.1126/sciadv.adf1785
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