Systematic functional analyses of spliceosomal components reveal novel mechanisme of alternative splicing regulation

Alternative splicing is an essential regulatory layer of gene expression that expands the coding potential of the genome in multicellular organisms. The spliceosome -the sophisticated machinery involved in intron removal- allows versatile regulation of gene expression programs. The splicing process...

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Detalles Bibliográficos
Autor: Tejedor Vaquero, Juan Ramón
Tipo de recurso: tesis doctoral
Estado:Versión publicada
Fecha de publicación:2014
País:España
Institución:CBUC, CESCA
Repositorio:TDR. Tesis Doctorales en Red
OAI Identifier:oai:www.tdx.cat:10803/385718
Acceso en línea:http://hdl.handle.net/10803/385718
Access Level:acceso abierto
Palabra clave:Alternative splicing
Anti-tumor drugs
Fas/CD95
Pre-mRNA splicing
Spliceosome
Regulatory networks
RNA binding
SR proteins
Zinc Finger
Empalmament alternatiu
Medicaments antitumorals
RNA missatger
Espliceosoma
Dit de zinc
Procesamiento alternativo
575
Descripción
Sumario:Alternative splicing is an essential regulatory layer of gene expression that expands the coding potential of the genome in multicellular organisms. The spliceosome -the sophisticated machinery involved in intron removal- allows versatile regulation of gene expression programs. The splicing process relies on the dynamic interplay between hundreds of components of the spliceosome, and the steps at which the complex process of the splicing reaction can be regulated remain largely unknown. The main objective of this thesis has been to develop high- throughput approaches to systematically identify novel regulators of alternative splicing, as well as to study the mechanisms by which they modulate splice site choice. We have identified a variety of regulators of Fas/CD95 alternative splicing within and outside of the splicing machinery and provide novel insights into connections between iron homeostasis and alternative splicing regulation. Using computational networks, we carried out a systematic functional analysis of the spliceosome components and their regulatory potential. Our results reveal the extensive regulatory plasticity of core spliceosome components throughout its assembly process. They also identified links between alternative splicing and iron homeostasis, providing a mechanism by which iron modulates alternative splicing through regulation of the RNA binding properties of a Zinc knuckle domain in the SR regulatory protein SRSF7. The results of this thesis highlight the value of high throughput technologies and network analyses to study complex molecular mechanisms, and unveils novel functional connections between the splicing machinery and other cellular processes.