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Vol. 12, Issue 11, 3703-3715, November 2001

Rab4 Regulates Formation of Synaptic-like Microvesicles from Early Endosomes in PC12 Cells

Heidi de Wit, Yael Lichtenstein,dagger Regis B. Kelly,dagger Hans J. Geuze, Judith Klumperman,Dagger §|| and Peter van der Sluijs§||

 Department of Cell Biology and Institute of Biomembranes, University Medical Center, 3584 CX Utrecht, The Netherlands and  Dagger Center for Biomedical Genetics, The Netherlands; and  dagger Department of Biochemistry and Biophysics, Hormone Research Institute, University of California, San Francisco, California 94143-0534

Early endosomes in PC12 cells are an important site for the formation of synaptic-like microvesicles and constitutive recycling vesicles. By immunogold electron microscopy, the small GTPase rab4 was localized to early endosomes and numerous small vesicles in the cell periphery and Golgi area of PC12 cells. Overexpression of GTPase-deficient Q67Lrab4 increased the number of early endosome-associated and cytoplasmic vesicles, whereas expression of GDP-bound S22Nrab4 significantly increased the length of early endosomal tubules. In parallel, Q67Lrab4 induced a shift in rab4, VAMP2, and TfR label from early endosomes to peripheral vesicles, whereas S22Nrab4 increased early endosome labeling of all three proteins. These observations were corroborated by early endosome budding assays. Together, our data document a thus far unrecognized role for rab4 in the formation of synaptic-like microvesicles and add to our understanding of the formation of constitutive recycling vesicles from early endosomes.

|| Corresponding authors. E-mail addresses: pvander{at}knoware.nl or j.klumperman{at}lab.azu.nl.

§ J.K. and P.vdS. contributed equally to this work.

Molecular Biology of the Cell
Vol. 12, 3703-3715, November 2001
Copyright © 2001 by The American Society for Cell Biology


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