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J. Biol. Chem., Vol. 278, Issue 33, 30597-30604, August 15, 2003

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RLIP, an Effector of the Ral GTPases, Is a Platform for Cdk1 to Phosphorylate Epsin during the Switch Off of Endocytosis in Mitosis^*

Carine Rossé  , Sébastien L'Hoste  , Nicolas Offner  ¶, André Picard ¶ and Jacques Camonis  ||

From the Institut Curie, INSERM U528, 26 rue d'Ulm, 75248 Paris cedex 05 and ^¶UMR 7628, Banyuls sur mer cedex 66651, France

The Ral signaling pathway is critically involved in Ras-dependent oncogenesis. One of its key actors, RLIP/RalBP1, which participates in receptor endocytosis during interphase, is also involved in mitotic processes when endocytosis is switched off. During mitosis, RLIP76 is located on the duplicated centrosomes and is required for their proper separation and movement to the poles. We have looked for actors that associate with RLIP during mitosis. We show here that RLIP/RalBP1 interacts with an active p34cdc2·cyclinB1 (cdk1) enzyme and that this interaction is crucial for the mitotic phosphorylation of Epsin that, once phosphorylated, is no longer competent for endocytosis. We show also that this latter phosphorylation is dependent on Ral signaling. We propose that RLIP/RalBP1 is used as a platform by the mitotic cdk1 to facilitate the phosphorylation of Epsin, which makes Epsin incompetent for endocytosis during mitosis, when endocytosis is switched off.

Received for publication, March 3, 2003 , and in revised form, May 28, 2003.

^* This work was supported in part by Grant CT-99-00875 from the European Union and Grant 5440 from the Association de Recherche sur le Cancer. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Supported by fellowships from the Ministere de l'Education Nationale et de la Recherche and from the Ligue Nationale contre le Cancer.

^|| To whom correspondence should be addressed. Tel.: 33-01-42-34-66-54; Fax: 33-01-42-34-66-50; E-mail: Jacques.Camonis{at}curie.fr.

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