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Evolución De Redes De Regulación De Micrornas De Primates”

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Evolución De Redes De Regulación De Micrornas De Primates” CENTRO DE INVESTIGACIÓN Y DE ESTUDIOS AVANZADOS DEL INSTITUTO POLITÉCNICO NACIONAL UNIDAD IRAPUATO Unidad de Genómica Avanzada. Departamento de Ingeniería Genética “Evolución de Redes de Regulación de microRNAs de Primates” TESIS Que presenta: IBT Gilberto Alejandro Álvarez Canales Para obtener el grado de: MAESTRO EN CIENCIAS en la especialidad de Biología Integrativa Directores de Tesis. Dr. Cei Abreu-Goodger Dr. Luis José Delaye Arredondo Irapuato, Guanajuato Enero, 2018 El presente trabajo fue realizado bajo la asesoría del Dr. Cei Abreu-Goodger y el Dr. Luis José Delaye Arredondo en los laboratorios de Genómica Computacional del RNA y de Genómica Evolutiva en la Unidad de Genómica Avanzada y el Departamento de Ingeniería Genética de CINVESTAV Unidad Irapuato en Guanajuato, México. Se agradece al Consejo Nacional de Ciencia y Tecnología, CONACYT por la beca otorgada durante el periodo de realización del presente trabajo. 2 AGRADECIMIENTOS Agradezco a los profesores Cei Abreu y Luis Delaye por su valiosa guía que me aportaron en este trabajo. Además por permitirme trabajar en un tema un poco alejado de su área de más experiencia, creo que eso hizo a esta etapa una de las experiencias más enriquecedoras de mi vida. Su asesoría fue excelente junto con el apoyo que me brindaron los profesores Sean Rovito y Moisés Santillán tanto en la parte evolutiva como en la parte de redes de regulación, respectivamente. El discutir estos temas junto con ellos me ha hecho aprender mucho sobre la manera de hacer ciencia. Agradezco al Posgrado de Biología Integrativa, a la Dra. Laila Partida y el Dr. Alfredo Herrero que fueron coordinadores del posgrado durante este periodo y a mis amigos de este posgrado Arely, los Danis, Diana, Fer, Gaby, Pavel y Víctor por todos esos momentos y buenas experiencias que pasamos juntos. Estoy muy feliz de conocerlos. También agradezco a todas las amistades que hice o mantuve aquí en CINVESTAV Irapuato, mis compañeros de casa, amigos del Fútbol, del Volley-ball y de reuniones para cenar y discutir sobre ciencia. A mis amigos de Clubes de ciencia Adrián, Manuel, Román, David, Diego, Andrés y Falcón que durante este período me volví a encontrar en distintos lugares, estoy muy feliz que sigamos creciendo como personas. A los grupos de los laboratorios en los que estuve. A Gaby, Roberto, Beto, Dago, Javo, Pablo, Sergio, Manu, Dani, Mich, Rosy y Obed por las valiosas discusiones durante las reuniones de grupo y por la asesoría durante la preparación del proyecto. A Araceli por todo el apoyo en el uso de Mazorka y la instalación de software. A mis amistades que hice en el MBL bajo la tutoría de mis grandes héroes durante la universidad: Rob Phillips y Hernán García. Gracias a todos ellos por esas buenas experiencias y estoy seguro que los volveré a encontrar en este camino de la ciencia. Muchas gracias a Alfred Simkin por su todo su apoyo y asesoría en la implementación y desarrollo de herramientas que utilicé. A mi familia que siempre estuvo apoyándome en todos momentos. Claramente la influencia de mis padres y hermanos sobre la curiosidad y respecto por la naturaleza me ha hecho llegar hasta aquí. Estoy muy agradecido con ellos por su ayuda para crecer como persona. A Tzitziki por este equipo que hemos formado, sigamos creciendo como lo hemos hecho. Gracias por todo tu gran apoyo en esta etapa de la vida, las buenas experiencias conociendo nuevos lugares y por los muchos retos que hemos superado. Doy muchas gracias al Consejo Nacional de Ciencia y Tecnología (CONACyT) por el apoyo de beca que me otorgaron durante la maestría durante el tiempo que realicé este proyecto. 3 Índice 1. Resumen .............................................................................................................................. 11 2. Introducción. ......................................................................................................................... 12 2.1 La importancia de los elementos reguladores en la evolución. ..................................... 12 2.2 Clases de elementos reguladores de la transcripción. ................................................. 22 2.3 ¿Cómo evolucionan las redes de regulación? Rasgos Generales................................ 34 2.4 Evolución de los primates ............................................................................................ 45 2.5 Preguntas para abordar mediante el estudio de la evolución de las redes de regulación genética ............................................................................................................ 49 3. Objetivos ............................................................................................................................... 50 3.1 Objetivo general ........................................................................................................... 50 3.2 Objetivos específicos ................................................................................................... 50 4 Hipótesis ................................................................................................................................ 51 5 Justificación ........................................................................................................................... 51 6 Métodos ................................................................................................................................. 51 6.1 Definición de las métricas de recableado ..................................................................... 51 6.2 Selección de las especies a analizar ............................................................................ 56 6.4 Selección de familias de microRNAs ............................................................................ 57 6.5 Análisis de recableado en 3’UTRs de Primates ............................................................ 58 6.6 Análisis de categorías funcionales. .............................................................................. 69 6.7 Conjunto de 3’UTRs de un alineamiento de mamíferos ............................................... 71 6.8 Complementación con datos de transcriptómica comparativa entre chimpancé y humano. ............................................................................................................................. 72 6.9 Uso de familias no conservadas de microRNAs ........................................................... 73 6.10 Radiación adaptativa de los cíclidos de Malawi. ......................................................... 73 6.11 Esquema Representativo de los métodos .................................................................. 76 6.12 Resumen general de la metodología .......................................................................... 77 7. Resultados ............................................................................................................................ 78 7.1 Evaluación de las métricas de recableado de redes, mediante simulación .................. 78 7.2 Análisis de las 3’ UTR de la superfamilia de los Hominoides ....................................... 82 4 7.3 Análisis de un conjunto alternativo de 3’ UTRs de los primates obtenidas de un alineamiento de mamíferos ................................................................................................ 94 7.4 Familias de microRNAs con distintos niveles de conservación .................................. 105 7.5 Conjunto de secuencias de regiones 3’UTRs del grupo de los Cíclidos del Lago Malawi ............................................................................................................................. 107 8. Discusión ............................................................................................................................ 113 8.1 Efecto en los valores de recableado por las secuencias de regiones 3’UTR utlizadas .......................................................................................................................... 113 8.2 Implicaciones de los métodos de predicción de blancos ............................................ 116 8.3 Implicaciones de los controles usados ....................................................................... 117 8. 4. Método con microRNA maduro en comparación con el método de k-meros ............ 118 8.5. Métrica de recableado de Shou en comparación con la métrica de Simkin ............... 118 8.6 Conservación en primates.......................................................................................... 120 8.7 Familias de microRNAs con tasas de recableado altas. ............................................. 120 8.8 Familias de microRNA con tasas de recableado bajas. .............................................. 123 8.9 Blancos con evidencia de selección. .......................................................................... 123 8.10 Discrepancia entre categorías funcionales obtenidas con el recableado por microRNAs y recableado por blanco ................................................................................ 126 8.11 Efecto de otros elementos de regulación en la evolución fenotípica ......................... 127 8.12 Aplicación en cíclidos ............................................................................................... 127 9. Conclusiones ...................................................................................................................... 129 10 Perspectivas .....................................................................................................................
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