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Generation and Purification of Human INO80 Chromatin Remodeling Complexes and Subcomplexes
Published on: October 23, 2014
Un complejo de remodelación de la cromatina que carga la cohesión en los cromosomas humanos
Mohamed-Ali Hakimi1, Daniel A Bochar, John A Schmiesing
1The Wistar Institute, 3601 Spruce Street, Philadelphia, Pennsylvania 19104, USA.
Nature
|August 29, 2002
Resumen
Este estudio identifica un nuevo complejo de remodelación de la cromatina humana que contiene SNF2h y componentes de cohesión. Este complejo facilita el acceso al ADN nucleosómico, crucial para las interacciones ADN-proteína y la segregación cromosómica.
Área de la Ciencia:
- Biología Molecular Biología Molecular
- La epigenética es la epigenética.
- Biología de la cromatina Biología de la cromatina
Sus antecedentes:
- La estructura de orden superior del ADN nucleosómico impide las interacciones proteína-ADN.
- El complejo de cohesión requiere acceso al ADN nucleosómico para la segregación cromosómica.
- Los factores de remodelación de la cromatina mejoran la accesibilidad del ADN.
Objetivo del estudio:
- Para aislar y caracterizar un complejo de remodelación de la cromatina humana.
- Para investigar la interacción entre el complejo de cohesina y los remodeladores de cromatina.
- Para aclarar el papel de este complejo en la accesibilidad y regulación del ADN.
Principales métodos:
- Aislamiento de un complejo humano que contiene ISWI (SNF2h).
- Co-inmunoprecipitación para estudiar las interacciones de las proteínas (hRAD21-SNF2h).
- Inmunoprecipitación de la cromatina para mapear los sitios de unión (hRAD21, SNF2h, Mi2) en las secuencias de Alu.
- Análisis de las modificaciones de la cola de histona y los efectos de la metilación del ADN.
Principales resultados:
- Se aisló un nuevo complejo compuesto por componentes de SNF2h, cohesin (hRAD21) y NuRD.
- Se confirmó la interacción directa entre hRAD21 y SNF2h.
- SNF2h, hRAD21 y Mi2 se unen específicamente a los elementos del ADN humano que contienen secuencias de Alu.
- Las modificaciones de la cola de histona y la metilación del ADN se correlacionan con la asociación del complejo SNF2h/cohesina con la cromatina.
- La evidencia sugiere que la actividad de la ATPasa SNF2h está involucrada en la carga de hRAD21 en la cromatina.
Conclusiones:
- Un nuevo complejo SNF2h/cohesina juega un papel en la accesibilidad de la cromatina.
- Este complejo está involucrado en la regulación de la asociación de la cohesión con la cromatina, influenciada por modificaciones epigenéticas.
- La actividad de la ATPasa SNF2h es crítica para la carga de la cromatina hRAD21, lo que afecta las interacciones ADN-proteína y la segregación cromosómica.
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