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Interactions of GGA3 with the ubiquitin sorting machinery

Nature Cell Biology volume 6, pages 244–251 (2004)Cite this article

Abstract

The Golgi-localized, γ-ear-containing, Arf-binding (GGA) proteins constitute a family of clathrin adaptors that are mainly associated with the trans-Golgi network (TGN)1,2,3 and mediate the sorting of mannose 6-phosphate receptors4,5,6. This sorting is dependent on the interaction of the VHS domain of the GGAs with acidic-cluster-dileucine signals in the cytosolic tails of the receptors4,5. Here we demonstrate the existence of another population of GGAs that are associated with early endosomes. RNA interference (RNAi) of GGA3 expression results in accumulation of the cation-independent mannose 6-phosphate receptor and internalized epidermal growth factor (EGF) within enlarged early endosomes. This perturbation impairs the degradation of internalized EGF, a process that is normally dependent on the sorting of ubiquitinated EGF receptors (EGFRs) to late endosomes. Protein interaction analyses show that the GGAs bind ubiquitin. The VHS and GAT domains of GGA3 are responsible for this binding, as well as for interactions with TSG101, a component of the ubiquitin-dependent sorting machinery. Thus, GGAs may have additional roles in sorting of ubiquitinated cargo.

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Figure 1: Localization of the GGAs to endosomes.
Figure 2: Depletion of GGA3 affects the localization of the CI-MPR.
Figure 3: Depletion of GGA3 inhibits EGF degradation.
Figure 4: Interaction of GGAs with ubiquitin and TSG101.
Figure 5: Interaction of GGA3 with ubiquitin is required for endosomal sorting.

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Acknowledgements

We thank X. Zhu and A. San Miguel for excellent technical assistance, and S. Cohen, P. Woodman, R. Kahn, and M. S. Robinson for kind gifts of reagents.

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  1. Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, 20892, Maryland, USA

    Rosa Puertollano & Juan S. Bonifacino

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Correspondence to Juan S. Bonifacino.

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The authors declare no competing financial interests.

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Puertollano, R., Bonifacino, J. Interactions of GGA3 with the ubiquitin sorting machinery. Nat Cell Biol 6, 244–251 (2004). https://doi.org/10.1038/ncb1106

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