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Published online 8 August 2005. doi:10.1083/jcb.200503054
The Rockefeller University Press, 0021-9525 $8.00
JCB, Volume 170, Number 4, 627-636
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Article

 An AP-1/clathrin coat plays a novel and essential role in forming the Weibel-Palade bodies of endothelial cells

Winnie W.Y. Lui-Roberts, Lucy M. Collinson, Lindsay J. Hewlett, Grégoire Michaux, and Daniel F. Cutler

Medical Research Council Laboratory of Molecular Cell Biology, Cell Biology Unit and Department of Biochemistry and Molecular Biology, University College London, London, WC1E 6BT, England, UK

Correspondence to D.F. Cutler: d.cutler{at}ucl.ac.uk

Clathrin provides an external scaffold to form small 50–100-nm transport vesicles. In contrast, formation of much larger dense-cored secretory granules is driven by selective aggregation of internal cargo at the trans-Golgi network; the only known role of clathrin in dense-cored secretory granules formation is to remove missorted proteins by small, coated vesicles during maturation of these spherical organelles. The formation of Weibel-Palade bodies (WPBs) is also cargo driven, but these are cigar-shaped organelles up to 5 µm long. We hypothesized that a cytoplasmic coat might be required to make these very different structures, and we found that new and forming WPBs are extensively, sometimes completely, coated. Overexpression of an AP-180 truncation mutant that prevents clathrin coat formation or reduced AP-1 expression by small interfering RNA both block WPB formation. We propose that, in contrast to other secretory granules, cargo aggregation alone is not sufficient to form immature WPBs and that an external scaffold that contains AP-1 and clathrin is essential.

L.J. Hewlett's present address is Division of Molecular Neuroendocrinology, National Institute for Medical Research, London, NW7 1AA, England, UK.

Abbreviations used in this paper: BFA, brefeldin A; CCV, clathrin-coated vesicle; HEK, human embryonic kidney; HUVEC, human umbilical vein endothelial cell; ISG, immature secretory granule; RNAi, RNA interference; siRNA, small interfering RNA; ssHRP, signal sequence HRP; VWF, von Willebrand factor; WPB, Weibel-Palade body.


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