CCDC32 stabilizes clathrin-coated pits and drives their invagination
Open accessPreprint — published as 10.7554/elife.107039 · Paper page
Abstract
Clathrin-mediated endocytosis (CME) is essential for maintaining homeostasis in mammalian cells. Previous studies have reported more than 50 CME accessory proteins; however, the mechanism driving the invagination of clathrin-coated pits (CCPs) remains elusive. We show by quantitative live cell imaging that siRNA-mediated knockdown of CCDC32, a poorly characterized endocytic accessory protein, leads to the accumulation of unstable flat clathrin assemblies. CCDC32 interacts with the α-appendage domain (AD) of AP2 in vitro and with full-length AP2 complexes in cells. Deletion of aa78-98 in CCDC32, corresponding to a predicted α-helix, abrogates AP2 binding and CCDC32’s early function in CME. Furthermore, clinically observed nonsense mutations in CCDC32, which result in C-terminal truncations that lack aa78-98, are linked to the development of cardio-facio-neuro-developmental syndrome (CFNDS). Overall, our data demonstrate the function of a novel endocytic accessory protein, CCDC32, in regulating CCP stabilization and invagination, critical early stages of CME.
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- Authors
- Yang, Z., Yang, C., Huang, Z., Xu, P., Li, Y., Han, L., Peng, L., Wei, X., Pak, J. E., Svitkina, T., Schmid, S. L., & Chen, Z.
- Affiliations
- University of South China (CN)
- University of Pennsylvania (US)
- Minjiang University (CN)
- The University of Texas Southwestern Medical Center (US)
- Corresponding author
- Chen, Zhiming
- Venue
- eLife
- Publisher
- eLife Sciences Publications, Ltd
- Published
- 2026-01-05
- Volume
- 14
- ISSN
- 2050-084X (ISSN-L)
- PMC
- PMC11230434
- OpenAlex
- W4400028319
- Cited by
- 3
- References
- 70
- Citations by year
- 2026: 2 · 2025: 1
- FWCI
- 6.73
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- eLife (publishedVersion, cc-by)
- bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion)
- PubMed (publishedVersion)
- PubMed Central (submittedVersion)
- Funders
- National Natural Science Foundation of China (32200564)
- Natural Science Foundation of Hunan Province (2024JJ2045)
- National Institutes of Health (R35 GM140832, GM73165)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
- Health Sciences › Medicine › Epidemiology › Autophagy in Disease and Therapy
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Caveolin-1 and cellular processes
- Keywords
- clathrin-mediated endocytosis, clathrin-coated pits, AP-2
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