ANTH domains within CALM, HIP1R, and Sla2 recognize ubiquitin internalization signals
- University of Iowa, United States
Abstract
Attachment of ubiquitin (Ub) to cell surface proteins serves as a signal for internalization via clathrin-mediated endocytosis (CME). How ubiquitinated membrane proteins engage the internalization apparatus remains unclear. The internalization apparatus contains proteins such as Epsin and Eps15, which bind Ub, potentially acting as adaptors for Ub-based internalization signals. Here we show that additional components of the endocytic machinery including CALM, HIP1R, and Sla2 bind Ub via their N-terminal ANTH domain, a domain belonging to the superfamily of ENTH and VHS domains. Structural studies revealed that Ub binds with µM affinity to a unique C-terminal region within the ANTH domain not found in ENTH domains. Functional studies showed that combined loss of Ub-binding by ANTH-domain proteins and other Ub-binding domains within the yeast internalization apparatus caused defects in the Ub-dependent internalization of the GPCR Ste2 that was engineered to rely exclusively on Ub as an internalization signal. In contrast, these mutations had no effect on the internalization of Ste2 engineered to use an alternate Ub-independent internalization signal. These studies define new components of the internalization machinery that work collectively with Epsin and Eps15 to specify recognition of Ub as an internalization signal.
Data availability
Diffraction data have been deposited in PDB under the accession code 7JXV.
Article and author information
Author details
Nicholas J Schnicker
Funding
National Institutes of Health (GM058202)
- Robert C Piper
National Institutes of Health (GM058202)
- Robert C Piper
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Reviewing Editor
- Wade Harper, Harvard Medical School, United States
Version history
- Received: July 28, 2021
- Accepted: November 24, 2021
- Accepted Manuscript published: November 25, 2021 (version 1)
- Version of Record published: December 6, 2021 (version 2)
- Version of Record updated: December 16, 2021 (version 3)
Copyright
© 2021, Pashkova et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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ANTH domains within CALM, HIP1R, and Sla2 recognize ubiquitin internalization signals
eLife 10:e72583.
https://doi.org/10.7554/eLife.72583
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