CryoEM of RUVBL1-RUVBL2-ZNHIT2, a complex that interacts with pre-mRNA-processing-splicing factor 8.

Biogenesis of the U5 small nuclear ribonucleoprotein (snRNP) is an essential and highly regulated process. In particular, PRPF8, one of U5 snRNP main components, requires HSP90 working in concert with R2TP, a cochaperone complex containing RUVBL1 and RUVBL2 AAA-ATPases, and additional factors that a...

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Autores: González-Corpas, Ana, Cabezudo, Sofía, López-Perrote, Andrés, Degliesposti, Gianluca, Zarzuela, Eduardo, Skehel, J Mark, Muñoz, Javier, Serna, Marina, Llorca Blanco, Oscar Antonio
Tipo de documento: artigo
Data de publicação:2022
País:España
Recursos:Instituto de Salud Carlos III (ISCIII)
Repositório:Repisalud
Idioma:inglês
OAI Identifier:oai:repisalud.isciii.es:20.500.12105/13716
Acesso em linha:http://hdl.handle.net/20.500.12105/13716
Access Level:Acceso aberto
Palavra-chave:ATPases Associated with Diverse Cellular Activities
Carrier Proteins
DNA Helicases
HEK293 Cells
Humans
Phosphoproteins
Protein Binding
RNA Precursors
RNA Splicing
RNA, Messenger
RNA-Binding Proteins
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repository_id_str
spelling CryoEM of RUVBL1-RUVBL2-ZNHIT2, a complex that interacts with pre-mRNA-processing-splicing factor 8.González-Corpas, AnaCabezudo, SofíaLópez-Perrote, AndrésDegliesposti, GianlucaZarzuela, EduardoSkehel, J MarkMuñoz, JavierSerna, MarinaLlorca Blanco, Oscar AntonioATPases Associated with Diverse Cellular ActivitiesCarrier ProteinsDNA HelicasesHEK293 CellsHumansPhosphoproteinsProtein BindingRNA PrecursorsRNA SplicingRNA, MessengerRNA-Binding ProteinsBiogenesis of the U5 small nuclear ribonucleoprotein (snRNP) is an essential and highly regulated process. In particular, PRPF8, one of U5 snRNP main components, requires HSP90 working in concert with R2TP, a cochaperone complex containing RUVBL1 and RUVBL2 AAA-ATPases, and additional factors that are still poorly characterized. Here, we use biochemistry, interaction mapping, mass spectrometry and cryoEM to study the role of ZNHIT2 in the regulation of the R2TP chaperone during the biogenesis of PRPF8. ZNHIT2 forms a complex with R2TP which depends exclusively on the direct interaction of ZNHIT2 with the RUVBL1-RUVBL2 ATPases. The cryoEM analysis of this complex reveals that ZNHIT2 alters the conformation and nucleotide state of RUVBL1-RUVBL2, affecting its ATPase activity. We characterized the interactions between R2TP, PRPF8, ZNHIT2, ECD and AAR2 proteins. Interestingly, PRPF8 makes a direct interaction with R2TP and this complex can incorporate ZNHIT2 and other proteins involved in the biogenesis of PRPF8 such as ECD and AAR2. Together, these results show that ZNHIT2 participates in the assembly of the U5 snRNP as part of a network of contacts between assembly factors required for PRPF8 biogenesis and the R2TP-HSP90 chaperone, while concomitantly regulating the structure and nucleotide state of R2TP.Oxford University PressInstituto de Salud Carlos IIIMinisterio de Ciencia e InnovaciónComunidad de MadridEuropean Regional Development Fund20222022-03-0320222022-02-0220222022-02-02journal articlehttp://purl.org/coar/resource_type/c_6501VoRhttp://purl.org/coar/version/c_970fb48d4fbd8a85info:eu-repo/semantics/articleapplication/pdfapplication/pdfhttp://hdl.handle.net/20.500.12105/13716reponame:Repisaludinstname:Instituto de Salud Carlos III (ISCIII)InglésengPID2020-114429RB-I00 Not available Not availableP2018 NMT4443 t Not availableopen accesshttp://purl.org/coar/access_right/c_abf2Atribución-NoComercial-CompartirIgual 4.0 Internacionalhttp://creativecommons.org/licenses/by-nc-sa/4.0/info:eu-repo/semantics/openAccessoai:repisalud.isciii.es:20.500.12105/137162026-06-12T12:43:37Z
dc.title.none.fl_str_mv CryoEM of RUVBL1-RUVBL2-ZNHIT2, a complex that interacts with pre-mRNA-processing-splicing factor 8.
title CryoEM of RUVBL1-RUVBL2-ZNHIT2, a complex that interacts with pre-mRNA-processing-splicing factor 8.
spellingShingle CryoEM of RUVBL1-RUVBL2-ZNHIT2, a complex that interacts with pre-mRNA-processing-splicing factor 8.
González-Corpas, Ana
ATPases Associated with Diverse Cellular Activities
Carrier Proteins
DNA Helicases
HEK293 Cells
Humans
Phosphoproteins
Protein Binding
RNA Precursors
RNA Splicing
RNA, Messenger
RNA-Binding Proteins
title_short CryoEM of RUVBL1-RUVBL2-ZNHIT2, a complex that interacts with pre-mRNA-processing-splicing factor 8.
title_full CryoEM of RUVBL1-RUVBL2-ZNHIT2, a complex that interacts with pre-mRNA-processing-splicing factor 8.
title_fullStr CryoEM of RUVBL1-RUVBL2-ZNHIT2, a complex that interacts with pre-mRNA-processing-splicing factor 8.
title_full_unstemmed CryoEM of RUVBL1-RUVBL2-ZNHIT2, a complex that interacts with pre-mRNA-processing-splicing factor 8.
title_sort CryoEM of RUVBL1-RUVBL2-ZNHIT2, a complex that interacts with pre-mRNA-processing-splicing factor 8.
dc.creator.none.fl_str_mv González-Corpas, Ana
Cabezudo, Sofía
López-Perrote, Andrés
Degliesposti, Gianluca
Zarzuela, Eduardo
Skehel, J Mark
Muñoz, Javier
Serna, Marina
Llorca Blanco, Oscar Antonio
author González-Corpas, Ana
author_facet González-Corpas, Ana
Cabezudo, Sofía
López-Perrote, Andrés
Degliesposti, Gianluca
Zarzuela, Eduardo
Skehel, J Mark
Muñoz, Javier
Serna, Marina
Llorca Blanco, Oscar Antonio
author_role author
author2 Cabezudo, Sofía
López-Perrote, Andrés
Degliesposti, Gianluca
Zarzuela, Eduardo
Skehel, J Mark
Muñoz, Javier
Serna, Marina
Llorca Blanco, Oscar Antonio
author2_role author
author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Instituto de Salud Carlos III
Ministerio de Ciencia e Innovación
Comunidad de Madrid
European Regional Development Fund
dc.subject.none.fl_str_mv ATPases Associated with Diverse Cellular Activities
Carrier Proteins
DNA Helicases
HEK293 Cells
Humans
Phosphoproteins
Protein Binding
RNA Precursors
RNA Splicing
RNA, Messenger
RNA-Binding Proteins
topic ATPases Associated with Diverse Cellular Activities
Carrier Proteins
DNA Helicases
HEK293 Cells
Humans
Phosphoproteins
Protein Binding
RNA Precursors
RNA Splicing
RNA, Messenger
RNA-Binding Proteins
description Biogenesis of the U5 small nuclear ribonucleoprotein (snRNP) is an essential and highly regulated process. In particular, PRPF8, one of U5 snRNP main components, requires HSP90 working in concert with R2TP, a cochaperone complex containing RUVBL1 and RUVBL2 AAA-ATPases, and additional factors that are still poorly characterized. Here, we use biochemistry, interaction mapping, mass spectrometry and cryoEM to study the role of ZNHIT2 in the regulation of the R2TP chaperone during the biogenesis of PRPF8. ZNHIT2 forms a complex with R2TP which depends exclusively on the direct interaction of ZNHIT2 with the RUVBL1-RUVBL2 ATPases. The cryoEM analysis of this complex reveals that ZNHIT2 alters the conformation and nucleotide state of RUVBL1-RUVBL2, affecting its ATPase activity. We characterized the interactions between R2TP, PRPF8, ZNHIT2, ECD and AAR2 proteins. Interestingly, PRPF8 makes a direct interaction with R2TP and this complex can incorporate ZNHIT2 and other proteins involved in the biogenesis of PRPF8 such as ECD and AAR2. Together, these results show that ZNHIT2 participates in the assembly of the U5 snRNP as part of a network of contacts between assembly factors required for PRPF8 biogenesis and the R2TP-HSP90 chaperone, while concomitantly regulating the structure and nucleotide state of R2TP.
publishDate 2022
dc.date.none.fl_str_mv 2022
2022-03-03
2022
2022-02-02
2022
2022-02-02
dc.type.none.fl_str_mv journal article
http://purl.org/coar/resource_type/c_6501
VoR
http://purl.org/coar/version/c_970fb48d4fbd8a85
dc.type.openaire.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.none.fl_str_mv http://hdl.handle.net/20.500.12105/13716
url http://hdl.handle.net/20.500.12105/13716
dc.language.none.fl_str_mv Inglés
eng
language_invalid_str_mv Inglés
language eng
dc.relation.none.fl_str_mv PID2020-114429RB-I00 Not available Not available
P2018 NMT4443 t Not available
dc.rights.none.fl_str_mv open access
http://purl.org/coar/access_right/c_abf2
Atribución-NoComercial-CompartirIgual 4.0 Internacional
http://creativecommons.org/licenses/by-nc-sa/4.0/
dc.rights.openaire.fl_str_mv info:eu-repo/semantics/openAccess
rights_invalid_str_mv open access
http://purl.org/coar/access_right/c_abf2
Atribución-NoComercial-CompartirIgual 4.0 Internacional
http://creativecommons.org/licenses/by-nc-sa/4.0/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Oxford University Press
publisher.none.fl_str_mv Oxford University Press
dc.source.none.fl_str_mv reponame:Repisalud
instname:Instituto de Salud Carlos III (ISCIII)
instname_str Instituto de Salud Carlos III (ISCIII)
reponame_str Repisalud
collection Repisalud
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15.198674

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