The mesenchymal morphology of cells expressing the EML4–ALK V3 oncogene is dependent on phosphorylation of Eg5 by NEK7

Echinoderm microtubule-associated protein-like 4 (EML4)–anaplastic lymphoma kinase (ALK) oncogenic fusion proteins are found in approximately 5% of non–small cell lung cancers. Different EML4–ALK fusion variants exist with variant 3 (V3) being associated with a significantly higher risk than other c...

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Autores: Pashley, Sarah L., Papageorgiou, Savvas, O'Regan, Laura, Barone, Giancarlo, Robinson, Susan W., Lucken, Kellie, Straatman, Kees R., Roig, Joan, Fry, Andrew M.
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2024
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/365776
Acceso en línea:http://hdl.handle.net/10261/365776
Access Level:acceso abierto
Palabra clave:EML4–ALK
NEK7
NEK9
Microtubules
Eg5
Eg5 inhibitors
NSCLC
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spelling The mesenchymal morphology of cells expressing the EML4–ALK V3 oncogene is dependent on phosphorylation of Eg5 by NEK7Pashley, Sarah L.Papageorgiou, SavvasO'Regan, LauraBarone, GiancarloRobinson, Susan W.Lucken, KellieStraatman, Kees R.Roig, JoanFry, Andrew M.EML4–ALKNEK7NEK9MicrotubulesEg5Eg5 inhibitorsNSCLCEchinoderm microtubule-associated protein-like 4 (EML4)–anaplastic lymphoma kinase (ALK) oncogenic fusion proteins are found in approximately 5% of non–small cell lung cancers. Different EML4–ALK fusion variants exist with variant 3 (V3) being associated with a significantly higher risk than other common variants, such as variant 1 (V1). Patients with V3 respond less well to targeted ALK inhibitors, have accelerated rates of metastasis, and have poorer overall survival. A pathway has been described downstream of EML4–ALK V3 that is independent of ALK catalytic activity but dependent on the NEK9 and NEK7 kinases. It has been proposed that assembly of an EML4–ALK V3–NEK9–NEK7 complex on microtubules leads to cells developing a mesenchymal-like morphology and exhibiting enhanced migration. However, downstream targets of this complex remain unknown. Here, we show that the microtubule-based kinesin, Eg5, is recruited to interphase microtubules in cells expressing EML4–ALK V3, whereas chemical inhibition of Eg5 reverses the mesenchymal morphology of cells. Furthermore, we show that depletion of NEK7 interferes with Eg5 recruitment to microtubules in cells expressing EML4–ALK V3 and cell length is reduced, but this is reversed by coexpression of a phosphomimetic mutant of Eg5, in a site, S1033, phosphorylated by NEK7. Intriguingly, we also found that expression of Eg5-S1033D led to cells expressing EML4–ALK V1 adopting a more mesenchymal-like morphology. Together, we propose that Eg5 acts as a substrate of NEK7 in cells expressing EML4–ALK V3 and Eg5 phosphorylation promotes the mesenchymal morphology typical of these cells.We thank the University of Leicester Core Biotechnology Services for providing access to the Advanced Imaging Facility (Research Resource Identifier: SCR_020967) and the Biotechnology and Biological Sciences Research Council for providing funding for the Zeiss Airyscan microscope (grant no.: BB/S019510/1 ). We are grateful to Jene Choi (Asan Medical Centre, Seoul) for providing BEAS-2B cells expressing EML4-ALK V1 and V3.Peer reviewedElsevierAmerican Society for Biochemistry and Molecular BiologyBiotechnology and Biological Sciences Research Council (UK)Wellcome TrustWorldwide Cancer ResearchAgencia Estatal de Investigación (España)Ministerio de Ciencia e Innovación (España)Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]202420242024info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionapplication/pdfhttp://hdl.handle.net/10261/365776reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Inglés#PLACEHOLDER_PARENT_METADATA_VALUE#info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2021-2023/PID2021-127045NB-I00The underlying dataset has been published as supplementary material of the article in the publisher platform at DOI https://doi.org/10.1016/j.jbc.2024.107144https://doi.org/10.1016/j.jbc.2024.107144Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/3657762026-05-22T06:33:51Z
dc.title.none.fl_str_mv The mesenchymal morphology of cells expressing the EML4–ALK V3 oncogene is dependent on phosphorylation of Eg5 by NEK7
title The mesenchymal morphology of cells expressing the EML4–ALK V3 oncogene is dependent on phosphorylation of Eg5 by NEK7
spellingShingle The mesenchymal morphology of cells expressing the EML4–ALK V3 oncogene is dependent on phosphorylation of Eg5 by NEK7
Pashley, Sarah L.
EML4–ALK
NEK7
NEK9
Microtubules
Eg5
Eg5 inhibitors
NSCLC
title_short The mesenchymal morphology of cells expressing the EML4–ALK V3 oncogene is dependent on phosphorylation of Eg5 by NEK7
title_full The mesenchymal morphology of cells expressing the EML4–ALK V3 oncogene is dependent on phosphorylation of Eg5 by NEK7
title_fullStr The mesenchymal morphology of cells expressing the EML4–ALK V3 oncogene is dependent on phosphorylation of Eg5 by NEK7
title_full_unstemmed The mesenchymal morphology of cells expressing the EML4–ALK V3 oncogene is dependent on phosphorylation of Eg5 by NEK7
title_sort The mesenchymal morphology of cells expressing the EML4–ALK V3 oncogene is dependent on phosphorylation of Eg5 by NEK7
dc.creator.none.fl_str_mv Pashley, Sarah L.
Papageorgiou, Savvas
O'Regan, Laura
Barone, Giancarlo
Robinson, Susan W.
Lucken, Kellie
Straatman, Kees R.
Roig, Joan
Fry, Andrew M.
author Pashley, Sarah L.
author_facet Pashley, Sarah L.
Papageorgiou, Savvas
O'Regan, Laura
Barone, Giancarlo
Robinson, Susan W.
Lucken, Kellie
Straatman, Kees R.
Roig, Joan
Fry, Andrew M.
author_role author
author2 Papageorgiou, Savvas
O'Regan, Laura
Barone, Giancarlo
Robinson, Susan W.
Lucken, Kellie
Straatman, Kees R.
Roig, Joan
Fry, Andrew M.
author2_role author
author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Biotechnology and Biological Sciences Research Council (UK)
Wellcome Trust
Worldwide Cancer Research
Agencia Estatal de Investigación (España)
Ministerio de Ciencia e Innovación (España)
Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
dc.subject.none.fl_str_mv EML4–ALK
NEK7
NEK9
Microtubules
Eg5
Eg5 inhibitors
NSCLC
topic EML4–ALK
NEK7
NEK9
Microtubules
Eg5
Eg5 inhibitors
NSCLC
description Echinoderm microtubule-associated protein-like 4 (EML4)–anaplastic lymphoma kinase (ALK) oncogenic fusion proteins are found in approximately 5% of non–small cell lung cancers. Different EML4–ALK fusion variants exist with variant 3 (V3) being associated with a significantly higher risk than other common variants, such as variant 1 (V1). Patients with V3 respond less well to targeted ALK inhibitors, have accelerated rates of metastasis, and have poorer overall survival. A pathway has been described downstream of EML4–ALK V3 that is independent of ALK catalytic activity but dependent on the NEK9 and NEK7 kinases. It has been proposed that assembly of an EML4–ALK V3–NEK9–NEK7 complex on microtubules leads to cells developing a mesenchymal-like morphology and exhibiting enhanced migration. However, downstream targets of this complex remain unknown. Here, we show that the microtubule-based kinesin, Eg5, is recruited to interphase microtubules in cells expressing EML4–ALK V3, whereas chemical inhibition of Eg5 reverses the mesenchymal morphology of cells. Furthermore, we show that depletion of NEK7 interferes with Eg5 recruitment to microtubules in cells expressing EML4–ALK V3 and cell length is reduced, but this is reversed by coexpression of a phosphomimetic mutant of Eg5, in a site, S1033, phosphorylated by NEK7. Intriguingly, we also found that expression of Eg5-S1033D led to cells expressing EML4–ALK V1 adopting a more mesenchymal-like morphology. Together, we propose that Eg5 acts as a substrate of NEK7 in cells expressing EML4–ALK V3 and Eg5 phosphorylation promotes the mesenchymal morphology typical of these cells.
publishDate 2024
dc.date.none.fl_str_mv 2024
2024
2024
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
Publisher's version
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/365776
url http://hdl.handle.net/10261/365776
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv #PLACEHOLDER_PARENT_METADATA_VALUE#
info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2021-2023/PID2021-127045NB-I00
The underlying dataset has been published as supplementary material of the article in the publisher platform at DOI https://doi.org/10.1016/j.jbc.2024.107144
https://doi.org/10.1016/j.jbc.2024.107144
Sí
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Elsevier
American Society for Biochemistry and Molecular Biology
publisher.none.fl_str_mv Elsevier
American Society for Biochemistry and Molecular Biology
dc.source.none.fl_str_mv reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC
instname:Consejo Superior de Investigaciones Científicas (CSIC)
instname_str Consejo Superior de Investigaciones Científicas (CSIC)
reponame_str DIGITAL.CSIC. Repositorio Institucional del CSIC
collection DIGITAL.CSIC. Repositorio Institucional del CSIC
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15,812429

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