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RalA Interacts Directly with the Arf-Responsive, PIP2-Dependent Phospholipase D1

https://doi.org/10.1006/bbrc.1997.6793Get rights and content

Abstract

RalA GTPase associates with a phospholipase D (PLD) that is activated in v-Src- and v-Ras-transformed cells. Two mammalian PLDs were recently cloned: PLD1, which is activated by Arf family GTPases and dependent upon phosphatidylinositol-4,5-bisphosphate (PIP2), and PLD2, which is also dependent upon PIP2, but not stimulated by Arf. Another PLD has been described that is stimulated by oleate. Evidence is provided that the RalA-assiciated PLD is PLD1. First, the PLD precipitated by RalA from murine fibroblasts was stimulated by Arf, dependent upon PIP2, and inhibited by oleate. Second, immobilized RalA precipitated PLD1 from sf9 insect cells overexpressing PLD1. Third, a series of RalA mutants precipitated PLD activity from both PLD1-expressing insect cells and murine fibroblasts with the same efficiency. And finally, immobilized RalA precipitated PLD1 from a purified PLD1 preparation. These data argue that RalA associates directly with the Arf-responsive, PIP2-dependent PLD1.

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    DG, diacylglycerolPEt, phosphatidylethanol; PA, phosphatidic acid; PC, phosphatidylcholine; PIP2, phosphatidylinositol-4,5-bisphosphate; PLD, phospholipase D;

    1

    Present address: The Rockefeller University, New York, NY 10021.

    2

    To whom correspondence should be addressed at the Department of Biological Sciences, Hunter College of The City University of New York, 695 Park Avenue, New York, NY 10021. Fax: (212) 772-5227. E-mail: [email protected].

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