SUMOylation regulates LKB1 localization and its oncogenic activity in liver cancer

BACKGROUND: Even though liver kinase B1 (LKB1) is usually described as a tumor suppressor in a wide variety of tissues, it has been shown that LKB1 aberrant expression is associated with bad prognosis in Hepatocellular Carcinoma (HCC). METHODS: Herein we have overexpressed LKB1 in human hepatoma cel...

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Autores: Zubiete Franco, Imanol, García Rodríguez, Juan Luis, Lopitz Otsoa, Fernando, Serrano Maciá, Marina, Simón Espinosa, Jorge, Fernández Tussy, Pablo, Barbier Torres, Lucía, Fernández Ramos, David, Gutiérrez de Juan, Virginia, López de Davalillo, Sergio, Carlevaris, Onintza, Beguiristain Gómez, Adolfo, Villa, Erica, Calvisi, Diego F., Martín Plágaro, César Augusto, Berra Ramírez, Miren Edurne, Aspichueta Celaá, Patricia, Beraza, Naiara, Varela Rey, Marta, Ávila, Matías A., Rodríguez, Manuel S., Mato, José M., Díaz Moreno, Irene, Díaz Quintana, Antonio, Delgado, Teresa C., Martínez Chantar, María Luz
Tipo de recurso: artículo
Fecha de publicación:2019
País:España
Institución:Universidad del País Vasco
Repositorio:Addi. Archivo Digital para la Docencia y la Investigación
OAI Identifier:oai:addi.ehu.eus:10810/32766
Acceso en línea:http://hdl.handle.net/10810/32766
Access Level:acceso abierto
Palabra clave:HCC
LKB1
SIRT1
STRADalpha
SUMO
activated protein-kinase
tumor-suppressor lkb1
glycine n-methyltransferase
hypoxia-inducible factor
hepatocellular-carcinoma
molecular-dynamics
SUMO conjugation
C-ZETA
AMPK
phosphorylation
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oai_identifier_str oai:addi.ehu.eus:10810/32766
network_acronym_str ES
network_name_str España
repository_id_str
dc.title.none.fl_str_mv SUMOylation regulates LKB1 localization and its oncogenic activity in liver cancer
title SUMOylation regulates LKB1 localization and its oncogenic activity in liver cancer
spellingShingle SUMOylation regulates LKB1 localization and its oncogenic activity in liver cancer
Zubiete Franco, Imanol
HCC
LKB1
SIRT1
STRADalpha
SUMO
activated protein-kinase
tumor-suppressor lkb1
glycine n-methyltransferase
hypoxia-inducible factor
hepatocellular-carcinoma
molecular-dynamics
SUMO conjugation
C-ZETA
AMPK
phosphorylation
title_short SUMOylation regulates LKB1 localization and its oncogenic activity in liver cancer
title_full SUMOylation regulates LKB1 localization and its oncogenic activity in liver cancer
title_fullStr SUMOylation regulates LKB1 localization and its oncogenic activity in liver cancer
title_full_unstemmed SUMOylation regulates LKB1 localization and its oncogenic activity in liver cancer
title_sort SUMOylation regulates LKB1 localization and its oncogenic activity in liver cancer
dc.creator.none.fl_str_mv Zubiete Franco, Imanol
García Rodríguez, Juan Luis
Lopitz Otsoa, Fernando
Serrano Maciá, Marina
Simón Espinosa, Jorge
Fernández Tussy, Pablo
Barbier Torres, Lucía
Fernández Ramos, David
Gutiérrez de Juan, Virginia
López de Davalillo, Sergio
Carlevaris, Onintza
Beguiristain Gómez, Adolfo
Villa, Erica
Calvisi, Diego F.
Martín Plágaro, César Augusto
Berra Ramírez, Miren Edurne
Aspichueta Celaá, Patricia
Beraza, Naiara
Varela Rey, Marta
Ávila, Matías A.
Rodríguez, Manuel S.
Mato, José M.
Díaz Moreno, Irene
Díaz Quintana, Antonio
Delgado, Teresa C.
Martínez Chantar, María Luz
author Zubiete Franco, Imanol
author_facet Zubiete Franco, Imanol
García Rodríguez, Juan Luis
Lopitz Otsoa, Fernando
Serrano Maciá, Marina
Simón Espinosa, Jorge
Fernández Tussy, Pablo
Barbier Torres, Lucía
Fernández Ramos, David
Gutiérrez de Juan, Virginia
López de Davalillo, Sergio
Carlevaris, Onintza
Beguiristain Gómez, Adolfo
Villa, Erica
Calvisi, Diego F.
Martín Plágaro, César Augusto
Berra Ramírez, Miren Edurne
Aspichueta Celaá, Patricia
Beraza, Naiara
Varela Rey, Marta
Ávila, Matías A.
Rodríguez, Manuel S.
Mato, José M.
Díaz Moreno, Irene
Díaz Quintana, Antonio
Delgado, Teresa C.
Martínez Chantar, María Luz
author_role author
author2 García Rodríguez, Juan Luis
Lopitz Otsoa, Fernando
Serrano Maciá, Marina
Simón Espinosa, Jorge
Fernández Tussy, Pablo
Barbier Torres, Lucía
Fernández Ramos, David
Gutiérrez de Juan, Virginia
López de Davalillo, Sergio
Carlevaris, Onintza
Beguiristain Gómez, Adolfo
Villa, Erica
Calvisi, Diego F.
Martín Plágaro, César Augusto
Berra Ramírez, Miren Edurne
Aspichueta Celaá, Patricia
Beraza, Naiara
Varela Rey, Marta
Ávila, Matías A.
Rodríguez, Manuel S.
Mato, José M.
Díaz Moreno, Irene
Díaz Quintana, Antonio
Delgado, Teresa C.
Martínez Chantar, María Luz
author2_role author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv HCC
LKB1
SIRT1
STRADalpha
SUMO
activated protein-kinase
tumor-suppressor lkb1
glycine n-methyltransferase
hypoxia-inducible factor
hepatocellular-carcinoma
molecular-dynamics
SUMO conjugation
C-ZETA
AMPK
phosphorylation
topic HCC
LKB1
SIRT1
STRADalpha
SUMO
activated protein-kinase
tumor-suppressor lkb1
glycine n-methyltransferase
hypoxia-inducible factor
hepatocellular-carcinoma
molecular-dynamics
SUMO conjugation
C-ZETA
AMPK
phosphorylation
description BACKGROUND: Even though liver kinase B1 (LKB1) is usually described as a tumor suppressor in a wide variety of tissues, it has been shown that LKB1 aberrant expression is associated with bad prognosis in Hepatocellular Carcinoma (HCC). METHODS: Herein we have overexpressed LKB1 in human hepatoma cells and by using histidine pull-down assay we have investigated the role of the hypoxia-related post-translational modification of Small Ubiquitin-related Modifier (SUMO)ylation in the regulation of LKB1 oncogenic role. Molecular modelling between LKB1 and its interactors, involved in regulation of LKB1 nucleocytoplasmic shuttling and LKB1 activity, was performed. Finally, high affinity SUMO binding entities-based technology were used to validate our findings in a pre-clinical mouse model and in clinical HCC. FINDINGS: We found that in human hepatoma cells under hypoxic stress, LKB1 overexpression increases cell viability and aggressiveness in association with changes in LKB1 cellular localization. Moreover, by using site-directed mutagenesis, we have shown that LKB1 is SUMOylated by SUMO-2 at Lys178 hampering LKB1 nucleocytoplasmic shuttling and fueling hepatoma cell growth. Molecular modelling of SUMO modified LKB1 further confirmed steric impedance between SUMOylated LKB1 and the STe20-Related ADaptor cofactor (STRADalpha), involved in LKB1 export from the nucleus. Finally, we provide evidence that endogenous LKB1 is modified by SUMO in pre-clinical mouse models of HCC and clinical HCC, where LKB1 SUMOylation is higher in fast growing tumors. INTERPRETATION: Overall, SUMO-2 modification of LKB1 at Lys178 mediates LKB1 cellular localization and its oncogenic role in liver cancer. FUND: This work was supported by grants from NIH (US Department of Health and Human services)-R01AR001576-11A1 (J.M.M and M.L.M-C.), Gobierno Vasco-Departamento de Salud 2013111114 (to M.L.M.-C), ELKARTEK 2016, Departamento de Industria del Gobierno Vasco (to M.L.M.-C), MINECO: SAF2017-87301-R and SAF2014-52097-R integrado en el Plan Estatal de Investigacion Cientifica y Tecnica y Innovacion 2013-2016 cofinanciado con Fondos FEDER (to M.L.M.-C and J.M.M., respectively), BFU2015-71017/BMC MINECO/FEDER, EU (to A.D.Q. and I.D.M.), BIOEF (Basque Foundation for Innovation and Health Research): EITB Maratoia BIO15/CA/014; Instituto de Salud Carlos III:PIE14/00031, integrado en el Plan Estatal de Investigacion Cientifica y Tecnica y Innovacion 2013-2016 cofinanciado con Fondos FEDER (to M.L.M.-C and J.M.M), Asociacion Espanola contra el Cancer (T.C.D, P·F-T and M.L.M-C), Daniel Alagille award from EASL (to T.C.D), Fundacion Cientifica de la Asociacion Espanola Contra el Cancer (AECC Scientific Foundation) Rare Tumor Calls 2017 (to M.L.M and M.A), La Caixa Foundation Program (to M.L.M), Programma di Ricerca Regione-Universita 2007-2009 and 2011-2012, Regione Emilia-Romagna (to E.V.), Ramon Areces Foundation and the Andalusian Government (BIO-198) (A.D.Q. and I.D.M.), ayudas para apoyar grupos de investigacion del sistema Universitario Vasco IT971-16 (P.A.), MINECO:SAF2015-64352-R (P.A.), Institut National du Cancer, FRANCE, INCa grant PLBIO16-251 (M.S.R.), MINECO - BFU2016-76872-R to (E.B.). Work produced with the support of a 2017 Leonardo Grant for Researchers and Cultural Creators, BBVA Foundation (M.V-R). Finally, Ciberehd_ISCIII_MINECO is funded by the Instituto de Salud Carlos III. We thank MINECO for the Severo Ochoa Excellence Accreditation to CIC bioGUNE (SEV-2016-0644). Funding sources had no involvement in study design; in the collection, analysis, and interpretation of data; in the writing of the report; and in the decision to submit the paper for publication.
publishDate 2019
dc.date.none.fl_str_mv 2019
2019
2019
dc.type.none.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.none.fl_str_mv http://hdl.handle.net/10810/32766
url http://hdl.handle.net/10810/32766
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv info:eu-repo/grantAgreement/MINECO/SAF2015-64352-R/
info:eu-repo/grantAgreement/MINECO/BFU2016-76872-R/
info:eu-repo/grantAgreement/MINECO/BFU2015-71017/BMC
https://www.sciencedirect.com/science/article/pii/S235239641830608X?via%3Dihub
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/3.0/es/
Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0)
Atribución-NoComercial-SinDerivadas 3.0 España
eu_rights_str_mv openAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by-nc-nd/3.0/es/
Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0)
Atribución-NoComercial-SinDerivadas 3.0 España
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Elsevier
publisher.none.fl_str_mv Elsevier
dc.source.none.fl_str_mv reponame:Addi. Archivo Digital para la Docencia y la Investigación
instname:Universidad del País Vasco
instname_str Universidad del País Vasco
reponame_str Addi. Archivo Digital para la Docencia y la Investigación
collection Addi. Archivo Digital para la Docencia y la Investigación
repository.name.fl_str_mv
repository.mail.fl_str_mv
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spelling SUMOylation regulates LKB1 localization and its oncogenic activity in liver cancerZubiete Franco, ImanolGarcía Rodríguez, Juan LuisLopitz Otsoa, FernandoSerrano Maciá, MarinaSimón Espinosa, JorgeFernández Tussy, PabloBarbier Torres, LucíaFernández Ramos, DavidGutiérrez de Juan, VirginiaLópez de Davalillo, SergioCarlevaris, OnintzaBeguiristain Gómez, AdolfoVilla, EricaCalvisi, Diego F.Martín Plágaro, César AugustoBerra Ramírez, Miren EdurneAspichueta Celaá, PatriciaBeraza, NaiaraVarela Rey, MartaÁvila, Matías A.Rodríguez, Manuel S.Mato, José M.Díaz Moreno, IreneDíaz Quintana, AntonioDelgado, Teresa C.Martínez Chantar, María LuzHCCLKB1SIRT1STRADalphaSUMOactivated protein-kinasetumor-suppressor lkb1glycine n-methyltransferasehypoxia-inducible factorhepatocellular-carcinomamolecular-dynamicsSUMO conjugationC-ZETAAMPKphosphorylationBACKGROUND: Even though liver kinase B1 (LKB1) is usually described as a tumor suppressor in a wide variety of tissues, it has been shown that LKB1 aberrant expression is associated with bad prognosis in Hepatocellular Carcinoma (HCC). METHODS: Herein we have overexpressed LKB1 in human hepatoma cells and by using histidine pull-down assay we have investigated the role of the hypoxia-related post-translational modification of Small Ubiquitin-related Modifier (SUMO)ylation in the regulation of LKB1 oncogenic role. Molecular modelling between LKB1 and its interactors, involved in regulation of LKB1 nucleocytoplasmic shuttling and LKB1 activity, was performed. Finally, high affinity SUMO binding entities-based technology were used to validate our findings in a pre-clinical mouse model and in clinical HCC. FINDINGS: We found that in human hepatoma cells under hypoxic stress, LKB1 overexpression increases cell viability and aggressiveness in association with changes in LKB1 cellular localization. Moreover, by using site-directed mutagenesis, we have shown that LKB1 is SUMOylated by SUMO-2 at Lys178 hampering LKB1 nucleocytoplasmic shuttling and fueling hepatoma cell growth. Molecular modelling of SUMO modified LKB1 further confirmed steric impedance between SUMOylated LKB1 and the STe20-Related ADaptor cofactor (STRADalpha), involved in LKB1 export from the nucleus. Finally, we provide evidence that endogenous LKB1 is modified by SUMO in pre-clinical mouse models of HCC and clinical HCC, where LKB1 SUMOylation is higher in fast growing tumors. INTERPRETATION: Overall, SUMO-2 modification of LKB1 at Lys178 mediates LKB1 cellular localization and its oncogenic role in liver cancer. FUND: This work was supported by grants from NIH (US Department of Health and Human services)-R01AR001576-11A1 (J.M.M and M.L.M-C.), Gobierno Vasco-Departamento de Salud 2013111114 (to M.L.M.-C), ELKARTEK 2016, Departamento de Industria del Gobierno Vasco (to M.L.M.-C), MINECO: SAF2017-87301-R and SAF2014-52097-R integrado en el Plan Estatal de Investigacion Cientifica y Tecnica y Innovacion 2013-2016 cofinanciado con Fondos FEDER (to M.L.M.-C and J.M.M., respectively), BFU2015-71017/BMC MINECO/FEDER, EU (to A.D.Q. and I.D.M.), BIOEF (Basque Foundation for Innovation and Health Research): EITB Maratoia BIO15/CA/014; Instituto de Salud Carlos III:PIE14/00031, integrado en el Plan Estatal de Investigacion Cientifica y Tecnica y Innovacion 2013-2016 cofinanciado con Fondos FEDER (to M.L.M.-C and J.M.M), Asociacion Espanola contra el Cancer (T.C.D, P·F-T and M.L.M-C), Daniel Alagille award from EASL (to T.C.D), Fundacion Cientifica de la Asociacion Espanola Contra el Cancer (AECC Scientific Foundation) Rare Tumor Calls 2017 (to M.L.M and M.A), La Caixa Foundation Program (to M.L.M), Programma di Ricerca Regione-Universita 2007-2009 and 2011-2012, Regione Emilia-Romagna (to E.V.), Ramon Areces Foundation and the Andalusian Government (BIO-198) (A.D.Q. and I.D.M.), ayudas para apoyar grupos de investigacion del sistema Universitario Vasco IT971-16 (P.A.), MINECO:SAF2015-64352-R (P.A.), Institut National du Cancer, FRANCE, INCa grant PLBIO16-251 (M.S.R.), MINECO - BFU2016-76872-R to (E.B.). Work produced with the support of a 2017 Leonardo Grant for Researchers and Cultural Creators, BBVA Foundation (M.V-R). Finally, Ciberehd_ISCIII_MINECO is funded by the Instituto de Salud Carlos III. We thank MINECO for the Severo Ochoa Excellence Accreditation to CIC bioGUNE (SEV-2016-0644). Funding sources had no involvement in study design; in the collection, analysis, and interpretation of data; in the writing of the report; and in the decision to submit the paper for publication.This work was supported by grants from NIH (US Department of Health and Human services)-R01AR001576-11A1 (J.M.M and M.L.M-C.), Gobierno Vasco-Departamento de Salud 2013111114 (to M.L.M.-C), ELKARTEK 2016, Departamento de Industria del Gobierno Vasco (to M.L.M.-C), MINECO: SAF2017-87301-R and SAF2014-52097-R integrado en el Plan Estatal de Investigacion Cientifica y Tecnica y Innovacion 2013-2016 cofinanciado con Fondos FEDER (to M.L.M.-C and J.M.M., respectively), BFU2015-71017/BMC MINECO/FEDER, EU (to A.D.Q. and I.D.M.), BIOEF (Basque Foundation for Innovation and Health Research): EITB Maratoia BIO15/CA/014; Instituto de Salud Carlos III: PIE14/00031, integrado en el Plan Estatal de Investigacion Cientifica y Tecnica y Innovacion 2013-2016 cofinanciado con Fondos FEDER (to M.L.M.-C and J.M.M), Asociacion Espanola contra el Cancer (T.C.D, P center dot F-T and M.L.M-C), Daniel Alagille award from EASL (to T.C.D), Fundacion Cientifica de la Asociacion Espanola Contra el Cancer (AECC Scientific Foundation) Rare Tumor Calls 2017 (to M.L.M and M.A), La Caixa Foundation Program (to M.L.M), Programma di Ricerca Regione-Universita 2007-2009 and 2011-2012, Regione Emilia-Romagna (to E.V.), Ramon Areces Foundation and the Andalusian Government (BIO-198) (A.D.Q. and I.D.M.), ayudas para apoyar grupos de investigacion del sistema Universitario Vasco IT971-16 (P.A.), MINECO: SAF2015-64352-R (P.A.), Institut National du Cancer, FRANCE, INCa grant PLBIO16-251 (M.S.R.), MINECO -BFU2016-76872-R to (E.B.). Work produced with the support of a 2017 Leonardo Grant for Researchers and Cultural Creators, BBVA Foundation (M.V-R). Finally, Ciberehd_ISCIII_MINECO is funded by the Instituto de Salud Carlos III. We thank MINECO for the Severo Ochoa Excellence Accreditation to CIC bioGUNE (SEV-2016-0644). Funding sources had no involvement in study design; in the collection, analysis, and interpretation of data; in the writing of the report; and in the decision to submit the paper for publication.Elsevier201920192019info:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10810/32766reponame:Addi. Archivo Digital para la Docencia y la Investigacióninstname:Universidad del País VascoInglésinfo:eu-repo/grantAgreement/MINECO/SAF2015-64352-R/info:eu-repo/grantAgreement/MINECO/BFU2016-76872-R/info:eu-repo/grantAgreement/MINECO/BFU2015-71017/BMChttps://www.sciencedirect.com/science/article/pii/S235239641830608X?via%3Dihubinfo:eu-repo/semantics/openAccesshttp://creativecommons.org/licenses/by-nc-nd/3.0/es/Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0)Atribución-NoComercial-SinDerivadas 3.0 Españaoai:addi.ehu.eus:10810/327662026-06-18T09:23:17Z
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