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In Vitro Ubiquitination and Deubiquitination Assays of Nucleosomal Histones
Published on: July 25, 2019
Regulación de la polimerización de actina dependiente de WASH y el tráfico de proteínas por ubiquitinación
Yi-Heng Hao1, Jennifer M Doyle, Saumya Ramanathan
1Department of Physiology, UT Southwestern Dallas, TX 75390, USA.
Cell
|March 5, 2013
Resumen
La ubiquitinación regula el transporte endosómico de proteínas. La ligasa MAGE-L2-TRIM27 se dirige a WASH, activando la nucleación de actina y permitiendo el transporte mediado por retrómero.
Área de la Ciencia:
- Biología celular Biología celular.
- Biología Molecular Biología Molecular
- La bioquímica es la bioquímica.
Sus antecedentes:
- El tráfico de proteínas endosómicas es crucial para la función celular y está implicado en la enfermedad.
- Los patógenos a menudo secuestran las vías endosómicas para su propio beneficio.
- El complejo retromero y el complejo regulador WASH son actores clave en el transporte endosómico.
Objetivo del estudio:
- Para aclarar el papel de la ubiquitinación en el tráfico de proteínas endosómicas.
- Para identificar ligasas específicas de ubiquitina involucradas en el transporte mediado por retrómero.
- Comprender los mecanismos moleculares que vinculan la ubiquitinación con la nucleación de actina en los endosomas.
Principales métodos:
- Investigó la localización de MAGE-L2-TRIM27 utilizando coinmunoprecipitación y microscopía.
- Se utilizó el knockdown de siRNA para evaluar el impacto de MAGE-L2-TRIM27 y Ube2O en el transporte de retrómeros.
- Se examinó el efecto de MAGE-L2-TRIM27 en la ubiquitinación de WASH y la nucleación de F-actina.
- Se analizaron las consecuencias de interrumpir la ubiquitinación WASH en el transporte endosómico.
Principales resultados:
- MAGE-L2-TRIM27 se localiza en los endosomas a través de la interacción con el complejo retrómero.
- La desactivación de MAGE-L2-TRIM27 o Ube2O perjudica el transporte mediado por retrómero.
- La actividad de la ubiquitina ligasa MAGE-L2-TRIM27 es esencial para la nucleación de F-actina mediada por WASH.
- MAGE-L2-TRIM27 media la ubiquitinación ligada a K63 de WASH en K220, que es crítica para la nucleación y el transporte de F-actina.
Conclusiones:
- MAGE-L2-TRIM27 juega un papel vital en el transporte endosómico retrógrado.
- La ubiquitinación ligada a K63 de WASH por MAGE-L2-TRIM27 es un paso regulador clave para la nucleación de actina endosómica.
- Este estudio identifica una nueva señal de activación para el complejo regulador WASH.
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