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Generation and Purification of Human INO80 Chromatin Remodeling Complexes and Subcomplexes
Published on: October 23, 2014
Estructura y topología de subunidades del remodelador de cromatina INO80 y su complejo de nucleosomas
Alessandro Tosi1, Caroline Haas, Franz Herzog
1Department of Biochemistry, Ludwig-Maximilian University, 81377 Munich, Germany; Gene Center, Ludwig-Maximilian University, 81377 Munich, Germany.
Cell
|September 17, 2013
Resumen
Determinamos la estructura del remodelador de cromatina INO80, revelando su arquitectura de múltiples subunidades y cómo se une a los nucleosomas. Esto proporciona un marco para comprender su papel en la regulación génica y la reparación del ADN.
Área de la Ciencia:
- Biología Molecular Biología Molecular
- Biología Estructural Biología Estructural
- La bioquímica es la bioquímica.
Sus antecedentes:
- Los remodeladores de cromatina INO80/SWR1 son grandes complejos de múltiples subunidades cruciales para la transcripción y el mantenimiento del genoma.
- Estos complejos facilitan los procesos celulares esenciales mediante el intercambio de variantes de histona como H2A y H2A.Z.
Objetivo del estudio:
- Para dilucidar la arquitectura tridimensional del complejo Saccharomyces cerevisiae INO80.
- Comprender las bases estructurales de la unión nucleosómica y las implicaciones funcionales para la remodelación de la cromatina.
Principales métodos:
- Un enfoque integrador de biología estructural que combina la microscopía crioectrónica, el enlace cruzado y la espectrometría de masas.
- Análisis bioquímicos para caracterizar las interacciones de las subunidades y la dinámica de unión de nucleosomas.
Principales resultados:
- Determinó la arquitectura general "en forma de embrión" de INO80, que comprende distintos módulos de cabeza, cuello, cuerpo y pies.
- Identificó la localización de subunidades clave, incluido el heterododecamero Rvb1/Rvb2 en la cabeza y las proteínas relacionadas con la actina (Arp) en el pie.
- Reveló conformaciones dinámicas abiertas y cerradas y mapeó el sitio de unión del nucleosoma a una superficie cóncava cerca de la región del cuello.
Conclusiones:
- Estableció un marco estructural detallado para el remodelador de cromatina INO80.
- Proporcionó información sobre el mecanismo de reconocimiento y remodelación del nucleosoma por este complejo proteico esencial.
- Estableció las bases para futuros estudios sobre la familia más amplia INO80 / SWR1 de remodeladores de la cromatina.
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Human DNA is almost two meters long. However, it is compressed inside a tiny nucleus measuring only a few microns in diameter. To make this degree of compaction possible, DNA is organized into several sequential levels so that it can fit into such a tiny space. The most compact form of DNA is a chromosome that can be seen under a microscope in a dividing cell.
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Nucleosomes are the DNA-histone complex, where the DNA strand is wound around the histone core. The histone core is an octamer containing two copies of H2A, H2B, H3, and H4 histone proteins.
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