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Caenorhabditis elegans auxilin: a J-domain protein essential for clathrin-mediated endocytosis in vivo

Nature Cell Biology volume 3, pages 215–219 (2001)Cite this article

Abstract

The budding of clathrin-coated vesicles is essential for protein transport. After budding, clathrin must be uncoated before the vesicles can fuse with other membranous structures. In vitro, the molecular chaperone Hsc70 uncoats clathrin-coated vesicles in an ATP-dependent process that requires a specific J-domain protein such as auxilin. However, there is little evidence that either Hsc70 or auxilin is essential in vivo. Here we show that C. elegans has a single auxilin homologue that is identical to mammalian auxilin in its in vitro activity. When RNA-mediated interference (RNAi) is used to inhibit auxilin expression in C. elegans, oocytes show markedly reduced receptor-mediated endocytosis of yolk protein tagged with green fluorescent protein (GFP). In addition, most of these worms arrest during larval development, exhibit defective distribution of GFP–clathrin in many cell types, and show a marked change in clathrin dynamics, as determined by fluorescence recovery after photobleaching (FRAP). We conclude that auxilin is required for in vivo clathrin-mediated endocytosis and development in C. elegans.

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Figure 1: CeAUX-53 polymerizes clathrin and supports Hsc70–clathrin interaction.
Figure 2: C. elegans auxilin(RNAi) phenotypes.
Figure 3: GFP–CHC fluorescence in wild-type and auxilin(RNAi) worms.
Figure 4: Different dynamics of GFP–clathrin in coelomocytes from wild-type and auxilin(RNAi) worms.

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Acknowledgements

We thank M. Krause for valuable assistance, Y. Xu for electron micrographs of the baskets, A. Fire for GFP plasmids, Y. Kohara for cDNA clones of C. elegans auxilin, and W. Przylecki for technical support. This work was supported in part by March of Dimes grant FY99-583 (to D.H.).

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  1. Tsvika Greener and Barth Grant: These authors contributed equally to this work

Authors and Affiliations

  1. Laboratory of Cell Biology, NHLBI, NIH, Bethesda, 20892-0301, Md

    Tsvika Greener, Xufeng Wu, Lois E. Greene & Evan Eisenberg

  2. Columbia University College of Physicians and Surgeons, Dept. of Biochemistry and Molecular Biophysics, NY, 10032, NY

    Barth Grant, Yinhua Zhang & David Hirsh

Authors
  1. Tsvika Greener
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  2. Barth Grant
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  3. Yinhua Zhang
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  4. Xufeng Wu
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  5. Lois E. Greene
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  7. Evan Eisenberg
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Correspondence to Evan Eisenberg.

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Greener, T., Grant, B., Zhang, Y. et al. Caenorhabditis elegans auxilin: a J-domain protein essential for clathrin-mediated endocytosis in vivo. Nat Cell Biol 3, 215–219 (2001). https://doi.org/10.1038/35055137

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