Functional implication of the calmodulin-binding domain of the adaptor proteína Grb7

Tutor de la Tesis Doctoral: Mario Vallejo.

Detalhes bibliográficos
Autor: García-Palmero, Irene
Formato: tesis doctoral
Fecha de publicación:2012
País:España
Recursos:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/78198
Acesso em linha:http://hdl.handle.net/10261/78198
Access Level:acceso abierto
id ES_d22f5cecdc09d7535ac0d5d202d933d0
oai_identifier_str oai:digital.csic.es:10261/78198
network_acronym_str ES
network_name_str España
repository_id_str
spelling Functional implication of the calmodulin-binding domain of the adaptor proteína Grb7García-Palmero, IreneTutor de la Tesis Doctoral: Mario Vallejo.The growth factor receptor bound protein 7 (Grb7) gives name to a mammalian adaptor protein family that also includes Grb10 and Grb14. As an adaptor protein, Grb7 mediates signal transduction events initiated by tyrosine kinase receptors present in the plasma membrane or other cytosolic kinases through its Src homology 2 (SH2) domain. Our laboratory previously described the presence of a calmodulin (CaM)‐binding domain (CaM‐BD) located in the proximal region of the pleckstrin homology (PH) domain of Grb7 and described that Grb7 was able to bind CaM in a Ca2+‐dependent manner both in vitro and in living cells. In this Thesis, we first described that the other Grb7 family members, Grb10 and Grb14, are also CaM‐binding proteins (CaM‐BP). To investigate the role that CaM plays in Grb7 functionality, we generated a deletion mutant lacking the CaM‐BD named Grb7Δ. We report the differential behavior in vitro and in vivo of cells expressing Grb7Δ, as compared to those expressing either its wild type counterpart or cells lacking expression of this adaptor protein. Our results showed that deletion of the CaM‐BD resulted in a lower cell migration rate, impaired cell adhesion ability to the extracellular matrix, and a lower proliferation rate. Additionally, we demonstrated that Grb7Δ was unable to localize in the nuclear fraction and that inhibition of CaM favored the nuclear localization of wild type Grb7. Therefore, we proposed that the CaM‐BD of Grb7 overlapped its nuclear localization sequence (NLS). Besides, we performed in vivo experiments to study the role of the CaM‐BD of Grb7 in tumor growth and tumor‐associated angiogenesis by implanting glioma C6 cells stably expressing EYFP, EYFP‐Grb7 or EYFP‐Grb7Δ in the brain of rats and monitoring the tumor growth and its associated angiogenesis using Magnetic Resonance Imaging (MRI). We found that tumors generated by EYFP‐Grb7Δ‐expressing cells were smaller and less vascularized than those grown from cells expressing its wild type EYFPGrb7 counterpart or EYFP‐expressing control cells, suggesting an anti‐angiogenic effect of the mutant protein. Finally, using co‐immunoprecipitation and Mass Spectrometry‐based proteomics, we described novel Grb7 and/or Grb7Δ binding partners. We validated by Western blot Hsc70 (heat shock cognate 70 protein) and caprin‐1 (cytoplasmic activation/proliferation‐associated protein‐1) as Grb7 and Grb7Δ associated partners, and Nedd4 (neural precursor cell expressed developmentally down‐regulated protein 4) as a Grb7 binding partner.Este trabajo ha sido realizado en el Departamento de Biología del Cáncer del Instituto de Investigaciones Biomédicas “Alberto Sols” (CSICUAM), gracias a una Ayuda a la Formación del Profesorado Universitario (FPU) del Ministerio de Educación.Peer ReviewedUniversidad Autónoma de MadridVillalobo, Antonio2013201320122013info:eu-repo/semantics/doctoralThesishttp://purl.org/coar/resource_type/c_db06http://hdl.handle.net/10261/78198reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Inglésinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/781982026-05-22T06:33:51Z
dc.title.none.fl_str_mv Functional implication of the calmodulin-binding domain of the adaptor proteína Grb7
title Functional implication of the calmodulin-binding domain of the adaptor proteína Grb7
spellingShingle Functional implication of the calmodulin-binding domain of the adaptor proteína Grb7
García-Palmero, Irene
title_short Functional implication of the calmodulin-binding domain of the adaptor proteína Grb7
title_full Functional implication of the calmodulin-binding domain of the adaptor proteína Grb7
title_fullStr Functional implication of the calmodulin-binding domain of the adaptor proteína Grb7
title_full_unstemmed Functional implication of the calmodulin-binding domain of the adaptor proteína Grb7
title_sort Functional implication of the calmodulin-binding domain of the adaptor proteína Grb7
dc.creator.none.fl_str_mv García-Palmero, Irene
author García-Palmero, Irene
author_facet García-Palmero, Irene
author_role author
dc.contributor.none.fl_str_mv Villalobo, Antonio
description Tutor de la Tesis Doctoral: Mario Vallejo.
publishDate 2012
dc.date.none.fl_str_mv 2012
2013
2013
2013
dc.type.none.fl_str_mv info:eu-repo/semantics/doctoralThesis
http://purl.org/coar/resource_type/c_db06
format doctoralThesis
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/78198
url http://hdl.handle.net/10261/78198
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Universidad Autónoma de Madrid
publisher.none.fl_str_mv Universidad Autónoma de Madrid
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
_version_ 1869420327907360768
score 15.81155

Registos relacionados