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In Vitro Ubiquitination and Deubiquitination Assays of Nucleosomal Histones
Published on: July 25, 2019
Estabilidad de los nucleosomas que contienen H2A y H2B ubicuitados homogéneamente preparados mediante semisíntesis
Beat Fierz1, Sinan Kilic, Aaron R Hieb
1Department of Chemistry, Princeton University, Princeton, New Jersey 08544, United States.
Journal of the American Chemical Society
|November 21, 2012
Resumen
Los investigadores desarrollaron un método para crear la histona H2A (uH2A) ubicuita para el estudio de la cromatina. Las histonas ubiquituladas tienen un modesto impacto directo en la estabilidad de los nucleosomas, al contrario de lo que se pensaba anteriormente.
Área de la Ciencia:
- La bioquímica es la bioquímica.
- Biología Molecular Biología Molecular
- La epigenética es la epigenética.
Sus antecedentes:
- Las modificaciones post-traducionales (PTM) de las histonas son cruciales para la regulación dinámica de la cromatina.
- Se supone que la ubicuidad de la histona afecta la estabilidad del nucleosoma, la unidad fundamental de la cromatina.
Objetivo del estudio:
- Desarrollar un método para la síntesis específica del sitio de la histona H2A ubicuitada (uH2A).
- Para investigar el impacto directo de la ubicuidad de histonas en la estabilidad de los nucleosomas.
Principales métodos:
- Semi-síntesis de uH2A utilizando un enfoque de ligación de proteínas expresadas en tres piezas.
- Incorporación de uH2A en los nucleosomas.
- Ensayos de estabilidad de nucleosomas acoplados a chaperonas usando uH2A y H2B. ubicuidad.
Principales resultados:
- Se ha generado con éxito una histona ubicuita específica del sitio H2A (uH2A) a través de la ligadura de proteínas expresadas.
- uH2A se puede incorporar de manera eficiente en los nucleosomas para futuros estudios.
- Las histonas ubiquituladas (H2A y H2B) exhiben un modesto efecto directo en la estabilidad nucleosómica.
Conclusiones:
- La estrategia de semisíntesis desarrollada permite estudios bioquímicos y biofísicos detallados de la ubicuidad de la histona.
- El impacto directo de la ubicuidad de histonas en la estabilidad de los nucleosomas es menos pronunciado de lo que se sugería anteriormente.
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02:10The Nucleosome Core Particle
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The paradox
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02:38Heterochromatin
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01:19The Nucleosome
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02:33The Nucleosome
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