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Necesidad de la prolyl isomerasa Pin1 para el punto de control de replicación

K E Winkler¹, K I Swenson, S Kornbluth

¹Department of Pharmacology and Cancer Biology, Duke University Medical Center, Box 3813, Durham, NC 27710, USA.

Science (New York, N.Y.)

|March 4, 2000

Resumen

La peptidil-prolyl isomerasa Pin1 es crucial para el punto de control de la replicación del ADN. Sin Pin1, las células entran prematuramente en mitosis, lo que indica su papel en retrasar la división celular hasta que se complete la replicación del ADN.

Área de la Ciencia:

Biología celular Biología celular.
Biología Molecular Biología Molecular
La bioquímica es la bioquímica.

Sus antecedentes:

La peptidil-prolyl isomerasa Pin1 juega un papel en la regulación del ciclo celular.
La función específica de Pin1 en los puntos de control de la replicación del ADN no se entiende completamente.

Objetivo del estudio:

Para investigar el papel de Pin1 en el punto de control de replicación del ADN en extractos de huevo de Xenopus laevis.
Para determinar si se requiere Pin1 para evitar la entrada mitótica prematura durante el estrés de replicación.

Principales métodos:

Los extractos de huevo de Xenopus laevis estaban agotados de Pin1.
La replicación del ADN fue inhibida usando aphidicolina.
Se agregaron Pin1 recombinante de tipo silvestre y un mutante inactivo para evaluar el rescate de la función de punto de control.
Se analizaron los niveles de fosforilación de Cdc25, la actividad de Cdc2 / ciclina B y los epítopos de MPM-2.

Principales resultados:

Los extractos empobrecidos con pin1 entraron en mitosis prematuramente cuando la replicación del ADN fue inhibida.
La adición de Pin1 de tipo salvaje restauró el punto de control de replicación, evitando la entrada mitótica prematura.
Un mutante Pin1 inactivo de la isomerasa no pudo rescatar el defecto del punto de control.
La mitosis prematura se asoció con la hiperfosforilación de Cdc25 y la activación de Cdc2 / ciclina B.

Conclusiones:

Pin1 es esencial para el punto de control de la replicación del ADN en Xenopus.
La actividad de Pin1 es necesaria para retrasar el inicio de la mitosis en respuesta a la replicación incompleta del ADN.
Pin1 probablemente funciona mediante la regulación de proteínas clave del ciclo celular como Cdc25.

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