STRUCTURAL BASES OF THE ACTION OF EIF3 AND OTHER FACTORS OF INITIATION OF EUKARYOTE TRANSLATION (Q3134784)
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Project Q3134784 in Spain
| Language | Label | Description | Also known as |
|---|---|---|---|
| English | STRUCTURAL BASES OF THE ACTION OF EIF3 AND OTHER FACTORS OF INITIATION OF EUKARYOTE TRANSLATION |
Project Q3134784 in Spain |
Statements
88,269.5 Euro
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176,539.0 Euro
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50.0 percent
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1 January 2018
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31 December 2020
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AGENCIA CONSEJO SUPERIOR DE INVESTIGACIONES CIENTIFICAS
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39°28'10.96"N, 0°22'34.82"W
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46250
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EL PROCESO DE TRADUCCION SE PUEDE DIVIDIR EN TRES ETAPAS: INICIACION, ELONGACION Y TERMINACION. A SU VEZ, EN EUCARIOTAS, LA INICIACION EN SU FORMA CANONICA SE PUEDE SUBDIVIDIR EN OCHO ETAPAS QUE IMPLICAN VARIOS COMPLEJOS ENTRE LAS SUBUNIDADES RIBOSOMALES Y LOS DIFERENTES FACTORES DE INICIACION (¿EUKARYOTIC INITIATION FACTORS; EIFS). HAY AL MENOS UNA DOCENA DE EIFS, DE LOS CUALES LOS MAS COMPLEJOS SON EIF3, EIF2 Y EIF4F. MIENTRAS QUE EIF2 Y EIF4F SON PROTEINAS HETEROTRIMERICAS, EIF3 SE COMPONE DE 6 SUBUNIDADES DIFERENTES EN SACCHAROMYCES CEREVISIAE, Y DE 13 SUBUNIDADES EN MAMIFEROS. _x000D_ LA ETAPA DE INICIACION DE LA TRADUCCION ES UN PROCESO ALTAMENTE CONTROLADO CUYA DESREGULACION SE ASOCIA A ENFERMEDADES COMO EL CANCER. POR EJEMPLO, EL ENSAMBLAJE Y LA ACTIVIDAD DE EIF4F, ASI COMO DE EIF2 Y EIF3 ESTAN FUERTEMENTE REGULADAS POR VIAS DE SEÑALIZACION INVOLUCRADAS EN PROCESOS CANCEROSOS, LO QUE ESTA DE ACUERDO CON LA DIVERSIDAD DE FUNCIONES DE ESTOS EIFS Y CON EL HECHO DE SON IMPORTANTES DIANAS PARA EL CONTROL TRADUCCIONAL. _x000D_ DURANTE MI PERIODO POSTDOCTORAL EN EL LABORATORIO DE VENKI RAMAKRISHNAN EN EL MRC-LMB (CAMBRIDGE) HEMOS CONTRIBUIDO, JUNTO AL TRABAJO DE OTROS DOS GRUPOS, A UNA MEJOR COMPRENSION ESTRUCTURAL DE LA INICIACION DE LA TRADUCCION. LA CARACTERIZACION DE CADA UNA DE LAS DIFERENTES ETAPAS DEPENDIA DE ATRAPAR ESTADOS FUNCIONALES PARTICULARES DE UN PROCESO MUY DINAMICO. ESTOS COMPLEJOS MACROMOLECULARES SON GRANDES Y HETEROGENEOS, Y PARA PODER OBTENER INFORMACION ESTRUCTURAL DE ELLOS APRENDI Y USE CRIO-MICROSCOPIA ELECTRONICA (CRIO-ME). SIN EMBARGO, Y A PESAR DE DETERMINAR VARIAS ESTRUCTURAS EN DIFERENTES ESTADOS FUNCIONALES Y DE DISPONER DE ESTRUCTURAS 3-D DE ALGUNOS EIFS, TODAVIA NO SE COMPRENDEN BIEN LAS FUNCIONES DE ESTOS ULTIMOS, SOBRE TODO DEBIDO A LA FALTA DE INFORMACION ESTRUCTURAL DETALLADA DE PARTES IMPORTANTES DE DICHOS FACTORES Y DE LOS COMPLEJOS EN QUE PARTICIPAN. EN PARTICULAR, SERIA MUY IMPORTANTE DETERMINAR LA ESTRUCTURA COMPLETA DE EIF3, EL MAS COMPLEJO DE LOS FACTORES DE INICIACION Y UN DIANA IMPORTANTE PARA LA REGULACION, Y LAS ESTRUCTURAS DE LOS COMPLEJOS DE ESTE FACTOR CON OTRAS PROTEINAS ALTAMENTE REGULADAS COMO EIF4F, EIF4B, EIF2, EIF5 Y EIF1._x000D_ CON ESTE PROPOSITO, TENGO LA INTENCION DE COMBINAR CRISTALOGRAFIA DE RAYOS X Y CRIO-ME PARA DILUCIDAR ESTAS ESTRUCTURAS, LO QUE ARROJARIA LUZ SOBRE ALGUNOS DE LOS ASPECTOS PARTICULARES DE LA INICIACION DE LA TRADUCCION EN EUCARIOTAS. GUIADOS POR LOS MAPAS DE CRYO-ME A BAJA RESOLUCION DISEÑAREMOS VARIAS CONSTRUCCIONES DE VARIOS DOMINIOS DE EIF3 Y LOS SOBRE-EXPRESAREMOS O CO-EXPRESAREMOS CONJUNTAMENTE CON OTROS EIFS O PARTES DE ELLOS EN SISTEMAS HETEROLOGOS Y LLEVAREMOS A CABO ESTUDIOS DE CRISTALIZACION DE RAYOS X. ACTUAREMOS DE FORMA SIMILAR PARA ELUCIDAR LA ESTRUCTURA DEL COMPLEJO EIF4F. DADO EL GRAN TAMAÑO DEL COMPLEJO MULTIFACTORIAL (EIF3-EIF2-EIF5-EIF2) USAREMOS ME, PRIMERO USANDO TINCION NEGATIVA Y MAS TARDE, USANDO CRIO-ME. A SU VEZ, EXPLORAREMOS LA POSIBILIDAD DE ESTUDIAR LA INTERACCION EIF3-EIF4F MEDIANTE ME SI OBTENEMOS COMPLEJOS LO SUFICIENTEMENTE ESTABLES._x000D_ POR ULTIMO, LA TECNICA DE CRIO-ME PUEDE SER TAMBIEN DE GRAN AYUDA PARA LA DETERMINACION ESTRUCTURAL DE OTRO TIPO DE COMPLEJOS MACROMOLECULARES Y POR TANTO, ME IMPLICARE EN EL DESARROLLO Y PROMOCION DE ESTA TECNICA QUE, DADO SU POTENCIAL, DEBERIA INTRODUCIRSE MAS AMPLIAMENTE EN VALENCIA (Y EN ESPAÑA), Y QUE PUEDE RESULTAR CRUCIAL PARA EL PROGRESO DE MUCHOS PROYECTOS ACTUALMENTE EN MARCHA EN EL IBV. (Spanish)
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TRANSLATION CAN BE ROUGHLY DIVIDED INTO THREE STAGES: INITIATION, ELONGATION AND TERMINATION. CANONICAL TRANSLATION INITIATION IN EUKARYOTES CAN BE SUBDIVIDED AS WELL INTO EIGHT STAGES INVOLVING SEVERAL COMPLEXES BETWEEN THE RIBOSOMAL SUBUNITS AND THE DIFFERENT INITIATION FACTORS (EIFS). THERE ARE (AT LEAST) A DOZEN OF INITIATION FACTORS, OF WHICH THE MOST COMPLEX ARE EIF3, EIF4F AND EIF2. WHILE EIF4F AND EIF2 ARE HETEROTRIMERIC PROTEINS, EIF3 IS COMPOSED OF 6 DIFFERENT SUBUNITS IN SACCHAROMYCES CEREVISIAE AND OF 13 SUBUNITS IN MAMMALS. _x000D_ PROTEIN SYNTHESIS IS A HIGHLY CONTROLLED PROCESS. IN FACT, DEREGULATION OF TRANSLATION INITIATION IN HUMANS IN ASSOCIATED WITH MANY DISEASES, INCLUDING CANCER. FOR EXAMPLE, ASSEMBLY/ACTIVITY OF EIF4F, AND OF EIF2 AND EIF3 COMPLEXES IS STRONGLY REGULATED BY SIGNALING PATHWAYS THAT ARE INVOLVED IN CANCER DEVELOPMENT AND ARE OFTEN OVEREXPRESSED IN HUMAN CANCERS, GIVING PRACTICAL INTEREST TO THE UNDERSTANDING OF THESE EIFS FUNCTIONS. DURING MY POSTDOCTORAL PERIOD IN THE VENKI RAMAKRISHNAN LAB., AT THE MRC-LMB (CAMBRIDGE), I HAVE CONTRIBUTED TO A BETTER UNDERSTANDING OF TRANSLATION INITIATION. THE CHARACTERIZATION OF EVERY ONE OF THE DIFFERENT STAGES DEPENDED ON TRAPPING PARTICULAR TRANSIENT FUNCTIONAL STATES OF A HIGHLY DYNAMIC PROCESS. THESE MACROMOLECULAR ASSEMBLIES ARE BIG AND HETEROGENEOUS AND WE LEARNT AND USED SINGLE-PARTICLE CRYO-EM TO OBTAIN SOME STRUCTURAL INFORMATION OF THEM. HOWEVER AND DESPITE THE RECENT DETERMINATION OF THESE STRUCTURES AND OF THE AVAILABILITY OF STRUCTURAL INFORMATION FOR SOME EUKARYOTIC INITIATION FACTORS, THE FUNCTIONS OF ALL THESE INITIATION FACTORS ARE STILL NOT WELL UNDERSTOOD, MAINLY DUE TO THE LACK OF DETAILED STRUCTURAL INFORMATION FOR IMPORTANT PARTS OF THESE FACTORS AND OF THE COMPLEXES FORMED BETWEEN THEM. IN PARTICULAR, IT WOULD BE HIGHLY IMPORTANT TO DETERMINE THE STRUCTURE OF COMPLETE EIF3, THE MOST COMPLEX OF THE EIFS AND AN IMPORTANT TARGET FOR REGULATION, AND THE STRUCTURES OF COMPLEXES OF EIF3 WITH OTHER IMPORTANT TARGETS FOR REGULATION SUCH AS EIF4F, EIF2, EIF5 AND EIF1. IN ORDER TO DO IT, I INTEND TO COMBINE X-RAY CRYSTALLOGRAPHY AND CRYO-EM TO ELUCIDATE THESE STRUCTURES, WHICH WOULD SHED LIGHT ON SOME PARTICULAR ASPECTS OF TRANSLATION INITIATION IN EUKARYOTES, AND PAVE THE WAY FOR A MECHANISTIC UNDERSTANDING OF HOW THE PROCESS CAN BE REGULATED. THIS KNOWLEDGE CAN PROVIDE US WITH A STRATEGY FOR TARGETING SOME COMPLEXES THAT CAN BE CANDIDATES FOR DEVELOPMENT OF POSSIBLE ANTI-TUMORAL TREATMENTS, AS WELL AS FOR DEVELOPMENT OF ANTIBIOTICS AND ANTIVIRALS._x000D_ BASED ON OUR PREVIOUS CRYO-EM MAPS AT LOW RESOLUTION, WE WILL DESIGN SEVERAL CONSTRUCTS OF DIFFERENT DOMAINS OF EIF3 AND OVEREXPRESS OR CO-EXPRESS THEM TOGETHER WITH OTHER EIFS OR PARTS OF THEM IN HETEROLOGOUS SYSTEMS AND WE WILL CARRY OUT X-RAY CRYSTALLIZATION TRIALS WITH THEM. WE WILL FOLLOW A SIMILAR APPROACH TO OBTAIN STRUCTURAL INFORMATION OF EIF4F. GIVEN THE LARGE SIZE OF THE MULTIFACTOR COMPLEX (EIF3-EIF2-EIF5-EIF2) WE WILL USE SINGLE PARTICLE EM TO DETERMINE ITS STRUCTURE. WE WILL FIRST USE NEGATIVE STAIN TECHNIQUES AND LATER ON CRYO-EM. IN ADDITION WE WILL EXPLORE THE SUITABILITY OF THE EIF3-EIF4F COMPLEX AS A TARGET FOR STRUCTURAL DETERMINATION BY EM IF WE ARE ABLE TO FORM A STABLE COMPLEX. FINALLY, I WILL BE INVOLVED IN PROMOTING AND DEVELOPING SINGLE PARTICLE EM IN VALENCIA.THIS IS A VERY POWERFUL TECHNIQUE THAT CAN BE CRUCIAL FOR THE SUCCESS OF THE ON-GOING RESEARCH OF MANY OTHER DIFFERENT GROUPS (English)
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Valencia
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Identifiers
BFU2017-85814-P
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