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Yeast epsin-related proteins required for Golgi–endosome traffic define a γ-adaptin ear-binding motif

Nature Cell Biology volume 5, pages 77–81 (2003)Cite this article

An Erratum to this article was published on 01 March 2003

Abstract

Clathrin-coated vesicles (CCVs) are a central component of endocytosis and traffic between the trans-Golgi network (TGN) and endosomes. Although endocytic CCV formation is well characterized, much less is known about CCV formation at internal membranes. Here we describe two epsin amino-terminal homology (ENTH) domain-containing proteins, Ent3p and Ent5p, that are intimately involved in clathrin function at the Golgi. Both proteins associate with the clathrin adaptor Gga2p in vivo; Ent5p also interacts with the clathrin adaptor complex AP-1 and clathrin. A novel, conserved motif that mediates the interaction of Ent3p and Ent5p with γ-ear domains of Gga2p and AP-1 is defined. Ent3p and Ent5p colocalize with clathrin, and cells lacking both Ent proteins exhibit defects in clathrin localization and traffic between the Golgi and endosomes. The findings suggest that Ent3p and Ent5p constitute a functionally related pair that co-operate with Gga proteins and AP-1 to recruit clathrin and promote formation of clathrin coats at the Golgi/endosomes. On the basis of our results and the established roles of epsin and epsin-related proteins in endocytosis, we propose that ENTH-domain-containing proteins are a universal component of CCV formation.

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Figure 1: Ent3p and Ent5p represent a conserved family of ENTH-domain-containing proteins.
Figure 2: Ent3p and Ent5p interact with the ear domains of Gga2p and AP-1-γ.
Figure 3: In vivo interactions of Ent3p and Ent5p with clathrin and adaptors.
Figure 4: Clathrin defects in ent3Δ ent5Δ cells.

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Acknowledgements

We are grateful to A. van der Bliek and D. Rube for help with the microscope, to S. Emr, C. Hirshberg, V. Kickhoeffer, and L. Silveira for antibodies, plasmids and strains. We thank B. Wendland and members of the Payne and van der Bliek lab for helpful discussions. This work was supported by National Institutes of Health grants GM39040 (G.S.P) and a fellowship from the Jonsson Cancer Center Foundation (M.D).

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  1. Mara C. Duncan and Giancarlo Costaguta: These authors contributed equally to this work

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  1. Department of Biological Chemistry, UCLA School of Medicine, Los Angeles, 90095, CA, USA

    Mara C. Duncan, Giancarlo Costaguta & Gregory S. Payne

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Correspondence to Gregory S. Payne.

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Duncan, M., Costaguta, G. & Payne, G. Yeast epsin-related proteins required for Golgi–endosome traffic define a γ-adaptin ear-binding motif. Nat Cell Biol 5, 77–81 (2003). https://doi.org/10.1038/ncb901

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