Crystal Structure of the Cyclostreptin-Tubulin Adduct: Implications for Tubulin Activation by Taxane-Site Ligands

It has been proposed that one of the mechanisms of taxane-site ligand-mediated tubulin activation is modulation of the structure of a switch element (the M-loop) from a disordered form in dimeric tubulin to a folded helical structure in microtubules. Here, we used covalent taxane-site ligands, inclu...

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Autores: Balaguer, Francisco de Asís, Mühlethaler, Tobias, Estévez-Gallego, Juan, Calvo, Enrique, Giménez-Abián, Juan F., Risinger, April L., Sorensen, Erik J., Vanderwal, Christopher D., Altmann, Karl-Heinz, Mooberry, Susan, Steinmetz, Michel O., Oliva, María A., Prota, Andrea E., Díaz, José Fernando
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2019
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/178957
Acceso en línea:http://hdl.handle.net/10261/178957
Access Level:acceso abierto
Palabra clave:Cyclostreptin
Tubulin
Microtubules
Multidrug resistance
Taxanes
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spelling Crystal Structure of the Cyclostreptin-Tubulin Adduct: Implications for Tubulin Activation by Taxane-Site LigandsBalaguer, Francisco de AsísMühlethaler, TobiasEstévez-Gallego, JuanCalvo, EnriqueGiménez-Abián, Juan F.Risinger, April L.Sorensen, Erik J.Vanderwal, Christopher D.Altmann, Karl-HeinzMooberry, SusanSteinmetz, Michel O.Oliva, María A.Prota, Andrea E.Díaz, José FernandoCyclostreptinTubulinMicrotubulesMultidrug resistanceTaxanesIt has been proposed that one of the mechanisms of taxane-site ligand-mediated tubulin activation is modulation of the structure of a switch element (the M-loop) from a disordered form in dimeric tubulin to a folded helical structure in microtubules. Here, we used covalent taxane-site ligands, including cyclostreptin, to gain further insight into this mechanism. The crystal structure of cyclostreptin-bound tubulin reveals covalent binding to βHis229, but no stabilization of the M-loop. The capacity of cyclostreptin to induce microtubule assembly compared to other covalent taxane-site agents demonstrates that the induction of tubulin assembly is not strictly dependent on M-loop stabilization. We further demonstrate that most covalent taxane-site ligands are able to partially overcome drug resistance mediated by βIII-tubulin (βIII) overexpression in HeLa cells, and compare their activities to pironetin, an interfacial covalent inhibitor of tubulin assembly that displays invariant growth inhibition in these cells. Our findings suggest a relationship between a diminished interaction of taxane-site ligands with βIII-tubulin and βIII tubulin-mediated drug resistance. This supports the idea that overexpression of βIII increases microtubule dynamicity by counteracting the enhanced microtubule stability promoted by covalent taxane-site binding ligands.This research was funded by Ministerio de Economia y Competitividad grant BFU2016-75319-R to JFDP (both AEI/FEDER, UE); Ministerio de Ciencia e Innovación RYC-2011-07900 to MAO; Swiss National Science Foundation grant (31003A_166608) to MOS and CA121138 to SLM. The authors acknowledge networking contribution by the COST Action CM1407 “Challenging organic syntheses inspired by nature”—from natural products chemistry to drug discoveryWe acknowledge support by the CSIC Open Access Publication Initiative through its Unit of Information Resources for Research (URICI)Peer reviewedMultidisciplinary Digital Publishing InstituteConsejo Superior de Investigaciones Científicas (España)Ministerio de Economía y Competitividad (España)Ministerio de Ciencia e Innovación (España)Swiss National Science FoundationEuropean CommissionConsejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]2019201920192019info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionhttp://hdl.handle.net/10261/178957reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Inglés#PLACEHOLDER_PARENT_METADATA_VALUE#info:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/BFU2016-75319-Rhttp://dx.doi.org/10.3390/ijms20061392Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/1789572026-05-22T06:33:51Z
dc.title.none.fl_str_mv Crystal Structure of the Cyclostreptin-Tubulin Adduct: Implications for Tubulin Activation by Taxane-Site Ligands
title Crystal Structure of the Cyclostreptin-Tubulin Adduct: Implications for Tubulin Activation by Taxane-Site Ligands
spellingShingle Crystal Structure of the Cyclostreptin-Tubulin Adduct: Implications for Tubulin Activation by Taxane-Site Ligands
Balaguer, Francisco de Asís
Cyclostreptin
Tubulin
Microtubules
Multidrug resistance
Taxanes
title_short Crystal Structure of the Cyclostreptin-Tubulin Adduct: Implications for Tubulin Activation by Taxane-Site Ligands
title_full Crystal Structure of the Cyclostreptin-Tubulin Adduct: Implications for Tubulin Activation by Taxane-Site Ligands
title_fullStr Crystal Structure of the Cyclostreptin-Tubulin Adduct: Implications for Tubulin Activation by Taxane-Site Ligands
title_full_unstemmed Crystal Structure of the Cyclostreptin-Tubulin Adduct: Implications for Tubulin Activation by Taxane-Site Ligands
title_sort Crystal Structure of the Cyclostreptin-Tubulin Adduct: Implications for Tubulin Activation by Taxane-Site Ligands
dc.creator.none.fl_str_mv Balaguer, Francisco de Asís
Mühlethaler, Tobias
Estévez-Gallego, Juan
Calvo, Enrique
Giménez-Abián, Juan F.
Risinger, April L.
Sorensen, Erik J.
Vanderwal, Christopher D.
Altmann, Karl-Heinz
Mooberry, Susan
Steinmetz, Michel O.
Oliva, María A.
Prota, Andrea E.
Díaz, José Fernando
author Balaguer, Francisco de Asís
author_facet Balaguer, Francisco de Asís
Mühlethaler, Tobias
Estévez-Gallego, Juan
Calvo, Enrique
Giménez-Abián, Juan F.
Risinger, April L.
Sorensen, Erik J.
Vanderwal, Christopher D.
Altmann, Karl-Heinz
Mooberry, Susan
Steinmetz, Michel O.
Oliva, María A.
Prota, Andrea E.
Díaz, José Fernando
author_role author
author2 Mühlethaler, Tobias
Estévez-Gallego, Juan
Calvo, Enrique
Giménez-Abián, Juan F.
Risinger, April L.
Sorensen, Erik J.
Vanderwal, Christopher D.
Altmann, Karl-Heinz
Mooberry, Susan
Steinmetz, Michel O.
Oliva, María A.
Prota, Andrea E.
Díaz, José Fernando
author2_role author
author
author
author
author
author
author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Consejo Superior de Investigaciones Científicas (España)
Ministerio de Economía y Competitividad (España)
Ministerio de Ciencia e Innovación (España)
Swiss National Science Foundation
European Commission
Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
dc.subject.none.fl_str_mv Cyclostreptin
Tubulin
Microtubules
Multidrug resistance
Taxanes
topic Cyclostreptin
Tubulin
Microtubules
Multidrug resistance
Taxanes
description It has been proposed that one of the mechanisms of taxane-site ligand-mediated tubulin activation is modulation of the structure of a switch element (the M-loop) from a disordered form in dimeric tubulin to a folded helical structure in microtubules. Here, we used covalent taxane-site ligands, including cyclostreptin, to gain further insight into this mechanism. The crystal structure of cyclostreptin-bound tubulin reveals covalent binding to βHis229, but no stabilization of the M-loop. The capacity of cyclostreptin to induce microtubule assembly compared to other covalent taxane-site agents demonstrates that the induction of tubulin assembly is not strictly dependent on M-loop stabilization. We further demonstrate that most covalent taxane-site ligands are able to partially overcome drug resistance mediated by βIII-tubulin (βIII) overexpression in HeLa cells, and compare their activities to pironetin, an interfacial covalent inhibitor of tubulin assembly that displays invariant growth inhibition in these cells. Our findings suggest a relationship between a diminished interaction of taxane-site ligands with βIII-tubulin and βIII tubulin-mediated drug resistance. This supports the idea that overexpression of βIII increases microtubule dynamicity by counteracting the enhanced microtubule stability promoted by covalent taxane-site binding ligands.
publishDate 2019
dc.date.none.fl_str_mv 2019
2019
2019
2019
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/10261/178957
url http://hdl.handle.net/10261/178957
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/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/BFU2016-75319-R
http://dx.doi.org/10.3390/ijms20061392
Sí
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Multidisciplinary Digital Publishing Institute
publisher.none.fl_str_mv Multidisciplinary Digital Publishing Institute
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 15,811543

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