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Regulación dinámica del empalme alternativo mediante silenciadores que modulan la competencia en el sitio de empalme
Yang Yu1, Patricia A Maroney, John A Denker
1Center for RNA Molecular Biology, Case Western Reserve University, Cleveland, OH 44106, USA.
Cell
|December 27, 2008
Resumen
Los investigadores identificaron nuevos motivos de secuencia que controlan el empalme alternativo al silenciar sitios específicos de empalme. Estos hallazgos mejoran nuestra comprensión de la regulación de la expresión génica y la diversidad proteómica.
Área de la Ciencia:
- Biología Molecular Biología Molecular
- Genética La genética.
- La bioquímica es la bioquímica.
Sus antecedentes:
- El empalme alternativo expande significativamente la diversidad proteómica en los eucariotas, con ~70% de los genes que producen múltiples isoformas.
- Los mecanismos moleculares precisos que rigen la selección del sitio de empalme permanecen en gran medida sin aclarar.
Objetivo del estudio:
- Para identificar elementos de secuencia que pueden regular la elección del sitio de empalme silenciando un sitio de empalme aguas abajo.
- Para investigar los mecanismos in vitro e in vivo del silenciamiento del sitio de empalme.
Principales métodos:
- Empleó un enfoque de selección por aleatorización in vitro para descubrir elementos de secuencia.
- Se probaron motivos identificados por su capacidad para silenciar un sitio de empalme proximal fuerte de 5'.
- Validaron los hallazgos in vivo e investigaron la base bioquímica del silenciamiento.
Principales resultados:
- Se identificaron seis nuevos motivos de secuencia (dos exónicos, cuatro intrónicos) que silencian efectivamente un sitio de empalme proximal de 5'.
- La actividad de silenciamiento dependía de la presencia de un sitio de empalme competidor aguas arriba.
- Los análisis bioquímicos sugieren que los motivos interfieren con la formación del complejo del sitio de empalme U1 snRNP/5', deteriorando el emparejamiento del sitio de empalme.
Conclusiones:
- Descubrió motivos de secuencia que actúan como reguladores del empalme alternativo mediante la modulación de la selección del sitio de empalme.
- Demostró que las perturbaciones en los pasos de empalme que no limitan la velocidad pueden influir profundamente en la elección del sitio de empalme.
- Proporciona nuevos conocimientos sobre los mecanismos que controlan la expresión génica y la diversidad proteómica.
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