[{"data":1,"prerenderedAt":-1},["ShallowReactive",2],{"detail-sidebar-cat-0-en-105":3,"doc-seo-440090-105":59,"doc-detail-440090-en":130},{"code":4,"msg":5,"data":6},0,"success",[7,13,18,23,28,33,38,43,48,51,55],{"id":8,"doc_module":4,"doc_module_name":9,"category_name":10,"show_sort_weight":11,"slug":12},1,"Document","Story & Novel",90,"story-novel",{"id":14,"doc_module":4,"doc_module_name":9,"category_name":15,"show_sort_weight":16,"slug":17},2,"Literature",80,"literature",{"id":19,"doc_module":4,"doc_module_name":9,"category_name":20,"show_sort_weight":21,"slug":22},4,"Exam",70,"exam",{"id":24,"doc_module":4,"doc_module_name":9,"category_name":25,"show_sort_weight":26,"slug":27},5,"Comic",60,"comic",{"id":29,"doc_module":4,"doc_module_name":9,"category_name":30,"show_sort_weight":31,"slug":32},6,"Technology",50,"technology",{"id":34,"doc_module":4,"doc_module_name":9,"category_name":35,"show_sort_weight":36,"slug":37},7,"Healthcare",40,"healthcare",{"id":39,"doc_module":4,"doc_module_name":9,"category_name":40,"show_sort_weight":41,"slug":42},8,"Research & Report",30,"research-report",{"id":44,"doc_module":4,"doc_module_name":9,"category_name":45,"show_sort_weight":46,"slug":47},9,"Religion & Spirituality",20,"religion-spirituality",{"id":46,"doc_module":4,"doc_module_name":9,"category_name":49,"show_sort_weight":46,"slug":50},"World Cup","world-cup",{"id":52,"doc_module":4,"doc_module_name":9,"category_name":53,"show_sort_weight":52,"slug":54},10,"Lifestyle","lifestyle",{"id":56,"doc_module":4,"doc_module_name":9,"category_name":57,"show_sort_weight":24,"slug":58},19,"General","general",{"code":4,"msg":60,"data":61},"ok",{"site_id":62,"language":63,"slug":64,"title":65,"keywords":66,"description":67,"schema_data":68,"social_meta":123,"head_meta":125,"extra_data":127,"updated_unix":129},105,"en","ccdc32-stabilizes-clathrin-coated-pits-and-drives-their-invagination-research-article","CCDC32 stabilizes clathrin-coated pits and drives their invagination - Research article","","Clathrin-mediated endocytosis (CME) is essential for mammalian cellular homeostasis, yet the mechanism that drives clathrin-coated pit invagination remains unclear. Quantitative live-cell imaging shows that siRNA knockdown of CCDC32 causes accumulation of unstable, flat clathrin assemblies. CCDC32 binds AP2 via the α-appendage domain in vitro and interacts with full-length AP2 complexes in cells. Deleting residues aa78-98 disrupts AP2 binding and early CME function, while C-terminal truncating nonsense mutations link CCDC32 loss to CFNDS.",{"@graph":69,"@context":122},[70,84,105],{"@type":71,"itemListElement":72},"BreadcrumbList",[73,77,79,82],{"item":74,"name":75,"@type":76,"position":8},"https://docshare.wps.com","Home","ListItem",{"item":78,"name":9,"@type":76,"position":14},"https://docshare.wps.com/document/",{"item":80,"name":40,"@type":76,"position":81},"https://docshare.wps.com/document/research-report/",3,{"item":83,"name":65,"@type":76,"position":19},"https://docshare.wps.com/document/ccdc32-stabilizes-clathrin-coated-pits-and-drives-their-invagination-research-article/440090/",{"url":83,"name":65,"@type":85,"image":86,"author":91,"headline":65,"publisher":94,"fileFormat":97,"inLanguage":63,"description":67,"dateModified":98,"datePublished":99,"encodingFormat":97,"isAccessibleForFree":100,"interactionStatistic":101},"DigitalDocument",{"url":87,"@type":88,"width":89,"height":90},"https://docshare.wps.com/thumbnails/ccdc32-stabilizes-clathrin-coated-pits-and-drives-their-invagination-research-article/440090.png","ImageObject",300,407,{"name":92,"@type":93},"Ben ","Person",{"url":74,"name":95,"@type":96},"DocShare","Organization","application/pdf","2026-09-30","2026-09-29",true,{"@type":102,"interactionType":103,"userInteractionCount":8},"InteractionCounter",{"@type":104},"ViewAction",{"@type":106,"mainEntity":107},"FAQPage",[108,114,118],{"name":109,"@type":110,"acceptedAnswer":111},"What effect does CCDC32 knockdown have on clathrin-coated pits?","Question",{"text":112,"@type":113},"siRNA-mediated CCDC32 knockdown leads to the accumulation of unstable, flat clathrin assemblies, indicating impaired progression toward productive invagination.","Answer",{"name":115,"@type":110,"acceptedAnswer":116},"How does CCDC32 interact with AP2?",{"text":117,"@type":113},"CCDC32 interacts with the α-appendage domain of AP2 in vitro and with full-length AP2 complexes in cells.",{"name":119,"@type":110,"acceptedAnswer":120},"Which CCDC32 region is essential for AP2 binding and early CME function?",{"text":121,"@type":113},"Deletion of amino acids 78–98 (a predicted α-helix) abolishes AP2 binding and eliminates CCDC32’s early role in clathrin-mediated endocytosis.","https://schema.org",{"og:url":83,"og:type":124,"og:title":65,"og:site_name":95,"og:description":67},"article",{"robots":126,"canonical":83},"index,follow",{"doc_id":128,"site_id":62},440090,1790793229,{"code":4,"msg":5,"data":131},{"doc_id":128,"user_id":132,"nickname":92,"user_avatar":133,"doc_module":4,"category_id":39,"category_name":40,"doc_title":65,"doc_description":67,"doc_content":134,"file_id":135,"file_url":136,"file_type":137,"file_size":138,"view_count":8,"is_deleted":4,"is_public":8,"is_downloadable":8,"audit_status":8,"page_count":139,"language":140,"language_code":63,"site_id":62,"html_lang":63,"table_of_contents":141,"faqs":142,"seo_title":143,"seo_description":67,"update_tm":144,"read_time":145},2336478951081,"https://ap-avatar.wpscdn.com/davatar_276721f389ce27ea32af1340a28f341c","RESEARCH ARTICLE  \n*For correspondence:  \n[sandra.schmid@czbiohub.org](sandra.schmid@czbiohub.org)[ ](sandra.schmid@czbiohub.org)(SLS);  \n[zhiming.chen@usc.edu.cn](zhiming.chen@usc.edu.cn) (ZC)  \nCompeting interest: The authors declare that no competing interests exist.  \nFunding: See page 19  \nSent for Review  \n25 April 2025  \nPreprint posted  \n29 April 2025  \nReviewed preprint posted  \n11 June 2025  \nReviewed preprint revised  \n15 August 2025  \nVersion of Record published  \n05 January 2026  \nReviewing Editor: Lei Lu, Nanyang Technological University, Singapore  \n Copyright Yang et al. This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use andredistribution provided that the original author and source are credited.  \nCCDC32 stabilizes clathrin-coated pitsand drives their invagination  \nZiyan Yang1, Changsong Yang2, Zheng Huang1, Peiliu Xu1, Yueping Li1, Lu Han1, Linyuan Peng1, Xiangying Wei3, John E Pak4, Tatyana Svitkina2,  \nSandra L Schmid4,5*, Zhiming Chen1*  \n1 NHC Key Laboratory of Birth Defect Research and Prevention, MOE Key Laboratory of Rare Pediatric Diseases, Institute of Cytology and Genetics of School of Basic Medical Sciences & Department of Clinical Laboratory of The First Affiliated Hospital, Hengyang Medical School, University of South China, Hengyang, China; 2 Department of Biology, University of Pennsylvania, Philadelphia, United States; 3 Fuzhou Institute of Oceanography, College of Geography and Oceanography, Minjiang University, Fuzhou, China; 4Chan Zuckerberg Biohub, San Francisco, United States; 5 Department of Cell Biology, University of Texas Southwestern Medical Center, Dallas, United States  \n\n| eLife Assessment\u003Cbr>The manuscript presents a valuable finding that CCDC32, beyond its reported role in AP2 assembly, follows AP2 to the plasma membrane and regulates clathrin-coated pit assembly and dynamics. The authors further identify an alpha-helical region within CCDC32 that is essential for its interaction with AP2 and its cellular function. While live-cell and ultrastructural imaging data are solid, future biochemical studies will be needed to confirm the proposed CCDC32-AP2 interaction.\u003Cbr>[Editors' note: this paper was reviewed by Review Commons.] |\n| --- |\n| 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. |\n\nIntroduction  \nClathrin-mediated endocytosis (CME) regulates nutrient uptake and maintains the activity of transmembrane transporters and is thus essential for maintaining cellular homeostasis (Kaksonen and Roux, 2018; Kirchhausen et al., 2014; McMahon and Boucrot, 2011; Mettlen et al., 2018) . Malfunctions of CME are strongly associated with neurological diseases, cardiovascular diseases, and cancers  \nYang et al. eLife 2025;14:RP107039. DOI: [https://doi.org/10.7554/eLife.107039](https://doi.org/10.7554/eLife.107039) 1 of 28  \n Research article Cell Biology  \n(Blue et al., 2018; DeMari et al., 2016; Elkin et al","cbCaikoxMaXyGRqF","https://ap.wps.com/l/cbCaikoxMaXyGRqF","pdf",10525738,28,"English","# Introduction\n## Clathrin-mediated endocytosis and clathrin-coated pit invagination\n## CCDC32 and cardio-facio-neuro-developmental syndrome (CFNDS)\n## Proposed CCDC32–AP2 functional connection","[{\"question\":\"What effect does CCDC32 knockdown have on clathrin-coated pits?\",\"answer\":\"siRNA-mediated CCDC32 knockdown leads to the accumulation of unstable, flat clathrin assemblies, indicating impaired progression toward productive invagination.\"},{\"question\":\"How does CCDC32 interact with AP2?\",\"answer\":\"CCDC32 interacts with the α-appendage domain of AP2 in vitro and with full-length AP2 complexes in cells.\"},{\"question\":\"Which CCDC32 region is essential for AP2 binding and early CME function?\",\"answer\":\"Deletion of amino acids 78–98 (a predicted α-helix) abolishes AP2 binding and eliminates CCDC32’s early role in clathrin-mediated endocytosis.\"}]","CCDC32 stabilizes clathrin-coated pits and drives their invagination - Research article | PDF",1790691068,71]