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Originally published In Press as doi:10.1074/jbc.M108508200 on October 3, 2001

J. Biol. Chem., Vol. 276, Issue 50, 47361-47370, December 14, 2001
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Inhibition of Transferrin Recycling and Endosome Tubulation by Phospholipase A2 Antagonists*

Paul de Figueiredo, Anne Doody, Renée S. Polizotto, Daniel Drecktrah, Salli Wood, Melanie Banta, Marian S. Strang, and William J. BrownDagger

From the Department of Molecular Biology and Genetics, Cornell University, Ithaca, New York 14853

We report here that a broad spectrum of phospholipase A2 (PLA2) antagonists produce a concentration-dependent, differential block in the endocytic recycling pathway of transferrin (Tf) and Tf receptors (TfRs) but have no acute affect on Tf uptake from the cell surface. At low concentrations of antagonists (~1 µM), Tf and TfR accumulated in centrally located recycling endosomes, whereas at higher concentrations (~10 µM), Tf-TfR accumulated in peripheral sorting endosomes. Several independent lines of evidence suggest that this inhibition of recycling may result from the inhibition of tubule formation. First, BFA-stimulated endosome tubule formation was similarly inhibited by PLA2 antagonists. Second, endocytosed tracers were found in larger spherical endosomes in the presence of PLA2 antagonists. And third, endosome tubule formation in a cell-free, cytosol-dependent reconstitution system was equally sensitive PLA2 antagonists. These results are consistent with the conclusion that endosome membrane tubules are formed by the action of a cytoplasmic PLA2 and that PLA2-dependent tubules are involved in intracellular recycling of Tf and TfR. When taken together with previous studies on the Golgi complex, these results also indicate that an intracellular PLA2 activity provides a novel molecular mechanism for inducing tubule formation from multiple organelles.

* This work was supported by National Institutes of Health Grant GM 60373 (to W. J. B.).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Dagger To whom correspondence should be addressed. Tel.: 607-255-2444; Fax: 607-255-2428; E-mail: wjb5@cornell.edu.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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