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