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B Un Funct Binding Niversi Depar Ional I G Doma Idad a Rtamen Mplica Ain Universidad Autónoma de Madrid Departamento de Bioquímica Functional implication of the calmodulin‐ binding domain of the adaptor protein Grb7 Irene García Palmero Madrid, 2012 Departamento de Bioquímica Facultad de Medicina Universidad Autónoma de Madrid Functional implication of the calmodulinn‐binding domain of the adaptor protein Grb7 Memoria de Tesis Doctoral presentada por: Irene García Palmero Licenciada en Bioquímica, para optar al grado de Doctor por la Universidad Autónoma de Madrid Director de tesis: Dr. Antonio Villalobo Profesor de Investigación del CSIC INSTITUTO DE INVESTIGACIONES BIOMÉDICAS “ALBERTO SOLS” CONSEJO SUPERIOR DE INVESTIGACIONES CIENTÍFICAS UNIVERSIDAD AUTÓNOMA DE MADRID Antonio Villalobo, Profesor de Investigación del Consejo Superior de Investigaciones Científicas y Profesor Honorario de la Universidad Autónoma de Madrid, certifica que: Irene García Palmero, Licenciada en Bioquímica por la Universidad Autónoma de Madrid, ha realizado bajo mi dirección el trabajo de investigación titulado: “Functional implication of the calmodulin‐binding domain of the adaptor protein Grb7” en el Instituto de Investigaciones Biomédicas “Alberto Sols”. Considero que el mencionado trabajo es apto para poder optar al grado de Doctor por la Universidad Autónoma de Madrid. Y para que así conste a todos los efectos, firmo el presente certificado en Madrid, a 12 de Junio de 2012. Fdo.: Antonio Villalobo Vº Bº Tutor: Mario Vallejo Director de la Tesis Profesor de Investigación, CSIC Profesor de Investigación, CSIC Profesor Honorario del Departamento de Bioquímica de la Universidad Autónoma de Madrid Este trabajo ha sido realizado en el Departamento de Biología del Cáncer del Instituto de Investigaciones Biomédicas “Alberto Sols” (CSIC‐ UAM), gracias a una Ayuda a la Formación del Profesorado Universitario (FPU) del Ministerio de Educación. A mis padres A mi hermana A David "Logic will get you from A to B. Imagination will take you everywhere" A. Einstein AGRADECIMIENTOS Parece que fue ayer cuando llegué por primera vez al IIB y la verdad es que han pasado ya casi 6 años. Realizar una Tesis doctoral ha sido un proyecto duro, pero una experiencia muy gratificante tanto a nivel profesional como personal, que he compartido con muchas personas a las que me gustaría dar las gracias de manera muy especial. En primer lugar quiero dar las gracias al Prof. Antonio Villalobo, mi director de Tesis, por haberme dado la oportunidad de incorporarme a su laboratorio y haber puesto a mi disposición todos sus conocimientos, por la importante labor de dirección, y sobre todo por el apoyo recibido durante estos años. También quiero dar las gracias a mi tutor, el Prof. Mario Vallejo por su labor en la supervisión de esta Tesis. Quiero agradecer también la ayuda que he recibido para el desarrollo de mis experimentos de MRI. En primer lugar al Prof. Sebastián Cerdán y a todo el equipo del SIERMAC y muy especialmente a la Dra. Pilar López Larrubia y a Patricia Sánchez, por su gran ayuda y apoyo en mis experimentos y en el buen desarrollo de esta tesis, y sobre todo por esos inolvidables momentos que hemos pasado jornaleando. También quiero dar las gracias al Dr. Eduardo Villalobo por la preparación del mutante de delección del dominio de unión a calmodulina Grb14∆, y a la Dra. Mª Carmen Grijota por su ayuda con la parte histológica de la Tesis. I would like to thank Prof. Jacques Haiech and his laboratory for performing the fluorescent polarization analysis and calorimetric titration assays to test the binding of calmodulin with the calmodulin‐binding domain peptides for us. También quiero dar las gracias a los servicios del IIB, cultivos, imagen, informática, biblioteca, secretaria, almacén, lavado, mantenimiento, prevención de riesgos laborales, microscopia, y a los laboratorios de confocal y citometría del SIDI y a toda la gente que hay detrás de ellos por sus buenas palabras y por su inestimable ayuda. Por la que ha sido mi segunda casa durante estos años, el labo 1.8, han pasado muchas personas con las que he compartido más o menos tiempo, pero que me han aportado y me han ayudado mucho a crecer como persona. Espero no olvidarme de nadie. En primer lugar, y por orden de llegada, me gustaría acordarme de Óscar, y de cómo él me enseñó todo lo que sabía durante mis inicios y de cómo yo le enseñé a reciclar. De Carmen y Elisa, y de el cerdo que pensábamos que iba a hacer estallar el IIB. De Amandeep y de Jihene, de esa persona con tan buen corazón, Karim, y de Célia, la dulzura francesa en persona. Gracias a Patri, por “estar completa”, a Christian por nuestras charlas sobre el Atleti, a Lucía, por enseñarme que las pipetas Pasteur de plástico también existen, a Lara por su gran apoyo en el labo y por su ayuda con el dicroísmo circular, y por supuesto a Miguel, con quien tanto me reí. A Celia muchas gracias por los panetones y por esos zuecos rojos de Papá Noel. También me quiero acordar de aquel verano polaco que pasé con Kate y con Michael, gracias por ser cómo eres y por considerarme “your Lady Grb7”. Como no, a mi Martita, terremoto y locura hecha persona, “muchas grassiass mi amol”. Gracias a mi Paulita, a la que yo intenté enseñar y resultó que ella me enseñó mucho más. Y no me quiero olvidar de otras personas que también han pasado por el labo como Juan, María, Ana, Gonzalo, Silviya, Clara, Laura y Cristina, mi guardiola preferida. Especialmente me gustaría recordar a mi compañero de tesis, de laboratorio, de día a día, Pablo, y dar las gracias a Marlies, esa belga que llegó revolucionando el laboratorio, por ser una persona tan especial a la que echo mucho de menos y a la que me quiero parecer de mayor. Pero sobre todo me gustaría dar las gracias a esa personita que es el alma del laboratorio, que tiene una luz tan especial y con olor a azahar, mi Amparito. Gracias por estar siempre ahí, por tu ayuda, por tus consejos, por nuestros cafés, risas, y sobre todo por haberme tratado siempre, desde que llegué al labo, con una vocecita que no oías, como a una hija. Y saliendo del 1.8, me quiero acordar especialmente de mi segundo cobijo, el 1.4. Muchas gracias en especial a Alberto, mi amigo, apoyo y compañero de siempre, a Isa, por toda esa locura y cariño que desbordas porque saben los chinos que voy a echar mucho de menos tus chistes y nuestros cotilleos desayunando, a Yenny, por su tranquilidad y buenas palabras. Quiero dar las gracias también a la Dra. Carmela Calés, por preocuparse siempre por mí durante estos años. No me quiero olvidar de gente que ya no están por ahí, como David, Rafa e Irma, me lo pasé genial con vosotros, y también de María, Dani, Camino, Bárbara y Leti. Quería dar las gracias a todas esas personas con las que he compartido pasillos, sonrisas y buenos momentos durante estos años. Los vecinos del 1.9, a Jose, Marina, Sandra, Diana y Elena, por sus risas, canciones de cumpleaños delante de toda la sala de personal y por alegrarme cada tarde que estaba en mi mesa cansada y me daba la vuelta para veros hacer carreras de sillas por el pasillo. También quiero agradecer todo el apoyo y el cariño recibido de los niños del 1.3.1, Jose y María V. Gracias Jose por hacerme reír cuando más lo he necesitado y a ti, María muchas gracias por tu apoyo durante esos momentos difíciles que he pasado en la biblio. Gracias a Jorge, mi arbitro de tenis preferido. Gracias también a Carlos y a Diego por alegrarme todos los días la entrada y salida del centro. No me puedo olvidar de gente con la que he compartido horas y horas en el cuarto de cultivos, como Jaime, Pilar, Rosa, Sole, María P, Bea, Cris y Álvaro. Ni tampoco de otras personas con las que he convivido estos últimos meses en la biblio, Dani, Ana y María G con quien he compartido agobios, estreses y sobre Anabel, muchas gracias por vuestros sabios consejos, y de Laura, ha sido un placer compartir cafecitos y confidencias durante estos meses. I want to say a big thank you to everybody who helped me during my aussie adventure. First of all I would like to thank Prof. Roger Daly for housing me in his lab and making me feel part of it since the first day. I also want to thank all the members of the lab, Naveid, for helping me with all the MS experiments, Brigid, Ling, Brian, Luxi, Carole and Emily for all your support during my stay. And special thanks to Ruth for all your help and good advice. I also would like to say thank you to the rest of the members of the lab 1 and to the cancer department, especially to Dessi and Christa, I miss you so much girls. Y todavía “down under”, no sé como agradecer a David y Fátima toda su ayuda y su apoyo durante esos meses. Muchas gracias por todo chicos. Y la verdad es que este largo camino de la tesis empezó hace mucho tiempo, allá por 2002, cuando decidí empezar biología en la Autónoma de Madrid, y conocí a mucha gente que me han ayudado a llegar donde estoy ahora. Cris, mi amiga, mi compañera de viajes y no sólo de tren, Laura, mi amiga, que siempre está ahí cuando la necesito, Marta, mi compañera de tesis, de biblio y de aventura, gracias por todo, Tefi, no cambies nunca, gracias por tu alegría, Tere los lunares hechos sonrisa, Dani y su empanamiento, mi Albertito (ver arriba) y mis niños Nacho y Sali.
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