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La hidroxilación de la asparagina del dominio de transactivación HIF es un interruptor hipoxico

David Lando¹, Daniel J Peet, Dean A Whelan

¹Department of Molecular Biosciences (Biochemistry), Adelaide University, SA 5005, Australia.

Science (New York, N.Y.)

|February 2, 2002

Resumen

Los factores inducibles a la hipoxia (HIF) ganan estabilidad y potencia en bajos niveles de oxígeno. Este estudio revela que el bloqueo de la hidroxilación de la asparagina, junto con la hidroxilación de la prolina, activa completamente los HIF al permitir la interacción con p300.

Área de la Ciencia:

Biología Molecular Biología Molecular
Respuesta Celular al Estrés Respuesta Celular al Estrés
Regulación genética Reglamento genético.

Sus antecedentes:

Los factores inducibles a la hipoxia (HIF) son factores de transcripción cruciales que regulan la adaptación celular a condiciones de bajo oxígeno.
La estabilidad y la actividad de HIF se controlan estrictamente, principalmente a través de la hidroxilación de prolina dirigida a HIF para la degradación a través del complejo de ligasa VHL-ubiquitina bajo normoxia.

Objetivo del estudio:

Para investigar el mecanismo de inducción hipóxica del dominio de transactivación terminal COOH (CAD) de HIF.
Determinar el papel de la hidroxilación de la asparagina dentro del HIF CAD en la regulación de la actividad transcripcional.

Principales métodos:

Se utilizan inhibidores de las dioxigenasas dependientes de Fe (II) y 2-oxoglutarato para bloquear la hidroxilación de la asparagina.
Mutagénesis dirigida por el sitio empleada, reemplazando la asparagina conservada (Asn) por alanina (Ala) en el CAD HIF.
Se evaluó la interacción entre HIF CAD y el coactivador de transcripción p300.

Principales resultados:

La inducción hipóxica de HIF CAD está mediada por la abrogación de la hidroxilación de la asparagina.
Los inhibidores dirigidos a las dioxigenasas impidieron la hidroxilación de Asn, facilitando la interacción de CAD con p300.
Se observó una interacción p300 constitutiva y una fuerte actividad transcripcional en el reemplazo de Asn por Ala.

Conclusiones:

La inducción completa de HIF-1alfa y HIF-2alfa requiere la abrogación simultánea de la hidroxilación de prolina y asparagina.
La hidroxilación de prolina regula la degradación de HIF, mientras que la hidroxilación de asparagina controla la actividad del dominio de transactivación bajo normoxia.

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