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Reclutamiento de un subcomplejo del proteasoma 19S a un promotor activado
Fernando Gonzalez1, Agnes Delahodde, Thomas Kodadek
1Center for Biomedical Inventions, University of Texas-Southwestern Medical Center, 5323 Harry Hines Boulevard, Dallas, TX 75390-8573, USA.
Science (New York, N.Y.)
|April 20, 2002
Resumen
El complejo proteasómico 19S, conocido por la descomposición de proteínas, también ayuda a la transcripción. Un subconjunto específico de proteínas 19S se une al promotor GAL1-10, lo que indica un papel no proteolítico en la transcripción de la ARN polimerasa II.
Área de la Ciencia:
- Biología Molecular Biología Molecular
- Biología celular Biología celular.
- La bioquímica es la bioquímica.
Sus antecedentes:
- La partícula reguladora del proteosoma 19S es conocida principalmente por su papel en la proteólisis celular.
- La evidencia emergente sugiere funciones no proteolíticas para las proteínas 19S en la reparación y transcripción del ADN.
Objetivo del estudio:
- Para investigar el papel de las proteínas del proteosoma 19S en la transcripción de la ARN polimerasa II.
- Para determinar si las proteínas 19S específicas son reclutadas para las regiones promotoras independientemente del complejo proteasómico completo.
Principales métodos:
- Se emplearon ensayos de inmunoprecipitación de cromatina (ChIP, por sus siglas en inglés) para detectar la unión de proteínas al promotor GAL1-10.
- El transactivador Gal4 se utilizó para inducir el reclutamiento de proteínas 19S.
- Se realizaron ensayos de retención de proteínas utilizando el dominio de activación Gal4 en extractos celulares.
Principales resultados:
- Las proteínas del complejo 19S son reclutadas para el promotor GAL1-10 por el transactivador Gal4.
- El complejo reclutado carece de la partícula proteolítica 20S y comprende un subconjunto de proteínas 19S.
- Este subconjunto específico de proteínas 19S es retenido por el dominio de activación Gal4 de extractos celulares.
Conclusiones:
- La base del complejo 19S puede funcionar independientemente de la maquinaria proteolítica más grande.
- Las proteínas 19S específicas juegan un papel directo y no proteolítico en la regulación de la transcripción de la ARN polimerasa II in vivo.
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