{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T14:55:45Z","timestamp":1790780145042,"version":"4.1.0"},"reference-count":40,"publisher":"Rockefeller University Press","issue":"12","license":[{"start":{"date-parts":[[2019,10,3]],"date-time":"2019-10-03T00:00:00Z","timestamp":1570060800000},"content-version":"vor","delay-in-days":736,"URL":"http:\/\/www.rupress.org\/terms\/"},{"start":{"date-parts":[[2018,6,4]],"date-time":"2018-06-04T00:00:00Z","timestamp":1528070400000},"content-version":"vor","delay-in-days":250,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc-sa\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100000265","name":"Medical Research Council","doi-asserted-by":"publisher","award":["MR\/J003964\/1"],"award-info":[{"award-number":["MR\/J003964\/1"]}],"id":[{"id":"10.13039\/501100000265","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003151","name":"Fonds de Recherche du Qu\u00e9bec - Nature et Technologies","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100003151","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["rupress.org"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2017,12,4]]},"abstract":"<jats:p>Clathrin-mediated endocytosis (CME) is the major route of receptor internalization at the plasma membrane. Analysis of constitutive CME is difficult because the initiation of endocytic events is unpredictable. When and where a clathrin-coated pit will form and what cargo it will contain are difficult to foresee. Here we describe a series of genetically encoded reporters that allow the initiation of CME on demand. A clathrin-binding protein fragment (\u201chook\u201d) is inducibly attached to an \u201canchor\u201d protein at the plasma membrane, which triggers the formation of new clathrin-coated vesicles. Our design incorporates temporal and spatial control by the use of chemical and optogenetic methods for inducing hook\u2013anchor attachment. Moreover, the cargo is defined. Because several steps in vesicle creation are bypassed, we term it \u201chot-wiring.\u201d We use hot-wired endocytosis to describe the functional interactions between clathrin and AP2. Two distinct sites on the \u03b22 subunit, one on the hinge and the other on the appendage, are necessary and sufficient for functional clathrin engagement.<\/jats:p>","DOI":"10.1083\/jcb.201702188","type":"journal-article","created":{"date-parts":[[2017,9,27]],"date-time":"2017-09-27T10:45:24Z","timestamp":1506509124000},"page":"4351-4365","update-policy":"https:\/\/doi.org\/10.1083\/jcb.crossmarkpolicy","source":"Crossref","is-referenced-by-count":55,"title":["New tools for \u201chot-wiring\u201d clathrin-mediated endocytosis with temporal and spatial precision"],"prefix":"10.1083","volume":"216","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-7450-2220","authenticated-orcid":true,"given":"Laura A.","family":"Wood","sequence":"first","affiliation":[{"name":"Centre for Mechanochemical Cell Biology, Warwick Medical School, University of Warwick, Coventry, England, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8295-9378","authenticated-orcid":true,"given":"Gabrielle","family":"Larocque","sequence":"additional","affiliation":[{"name":"Centre for Mechanochemical Cell Biology, Warwick Medical School, University of Warwick, Coventry, England, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3297-8604","authenticated-orcid":true,"given":"Nicholas I.","family":"Clarke","sequence":"additional","affiliation":[{"name":"Centre for Mechanochemical Cell Biology, Warwick Medical School, University of Warwick, Coventry, England, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0011-6700","authenticated-orcid":true,"given":"Sourav","family":"Sarkar","sequence":"additional","affiliation":[{"name":"Centre for Mechanochemical Cell Biology, Warwick Medical School, University of Warwick, Coventry, England, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8927-6967","authenticated-orcid":true,"given":"Stephen J.","family":"Royle","sequence":"additional","affiliation":[{"name":"Centre for Mechanochemical Cell Biology, Warwick Medical School, University of Warwick, Coventry, England, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"291","published-online":{"date-parts":[[2017,9,27]]},"reference":[{"key":"2023072309062837400_bib1","doi-asserted-by":"publisher","first-page":"279","DOI":"10.1016\/j.devcel.2013.06.019","article-title":"Advances in analysis of low signal-to-noise images link dynamin and AP2 to the functions of an endocytic checkpoint","volume":"26","author":"Aguet","year":"2013","journal-title":"Dev. Cell."},{"key":"2023072309062837400_bib2","doi-asserted-by":"publisher","first-page":"141","DOI":"10.1083\/jcb.201111049","article-title":"Multivariate proteomic profiling identifies novel accessory proteins of coated vesicles","volume":"197","author":"Borner","year":"2012","journal-title":"J. Cell Biol."},{"key":"2023072309062837400_bib3","doi-asserted-by":"publisher","first-page":"2102","DOI":"10.1242\/jcs.124834","article-title":"Specific removal of TACC3-ch-TOG-clathrin at metaphase deregulates kinetochore fiber tension","volume":"126","author":"Cheeseman","year":"2013","journal-title":"J. Cell Sci."},{"key":"2023072309062837400_bib4","doi-asserted-by":"publisher","first-page":"634","DOI":"10.1038\/ncb2478","article-title":"Reconstitution of clathrin-coated bud and vesicle formation with minimal components","volume":"14","author":"Dannhauser","year":"2012","journal-title":"Nat. Cell Biol."},{"key":"2023072309062837400_bib5","doi-asserted-by":"publisher","first-page":"431","DOI":"10.1126\/science.280.5362.431","article-title":"Association of the AP-3 adaptor complex with clathrin","volume":"280","author":"Dell\u2019Angelica","year":"1998","journal-title":"Science."},{"key":"2023072309062837400_bib6","doi-asserted-by":"publisher","first-page":"329","DOI":"10.1016\/j.devcel.2006.01.016","article-title":"Molecular switches involving the AP-2 beta2 appendage regulate endocytic cargo selection and clathrin coat assembly","volume":"10","author":"Edeling","year":"2006","journal-title":"Dev. Cell."},{"key":"2023072309062837400_bib7","doi-asserted-by":"publisher","first-page":"591","DOI":"10.1016\/j.cell.2004.08.017","article-title":"Endocytosis by random initiation and stabilization of clathrin-coated pits","volume":"118","author":"Ehrlich","year":"2004","journal-title":"Cell."},{"key":"2023072309062837400_bib8","doi-asserted-by":"publisher","first-page":"6572","DOI":"10.1073\/pnas.1117401109","article-title":"Clathrin-mediated endocytosis is inhibited during mitosis","volume":"109","author":"Fielding","year":"2012","journal-title":"Proc. Natl. Acad. Sci. USA."},{"key":"2023072309062837400_bib9","doi-asserted-by":"publisher","first-page":"773","DOI":"10.1016\/j.neuron.2006.08.029","article-title":"Clathrin-mediated endocytosis is the dominant mechanism of vesicle retrieval at hippocampal synapses","volume":"51","author":"Granseth","year":"2006","journal-title":"Neuron."},{"key":"2023072309062837400_bib10","doi-asserted-by":"publisher","first-page":"2185","DOI":"10.1242\/jcs.046177","article-title":"Functional equivalence of the clathrin heavy chains CHC17 and CHC22 in endocytosis and mitosis","volume":"122","author":"Hood","year":"2009","journal-title":"J. Cell Sci."},{"key":"2023072309062837400_bib11","doi-asserted-by":"publisher","first-page":"463","DOI":"10.1083\/jcb.201211127","article-title":"Coordination of adjacent domains mediates TACC3-ch-TOG-clathrin assembly and mitotic spindle binding","volume":"202","author":"Hood","year":"2013","journal-title":"J. Cell Biol."},{"key":"2023072309062837400_bib12","doi-asserted-by":"publisher","first-page":"739","DOI":"10.1083\/jcb.200911091","article-title":"Quantitative proteomics combined with BAC TransgeneOmics reveals in vivo protein interactions","volume":"189","author":"Hubner","year":"2010","journal-title":"J. Cell Biol."},{"key":"2023072309062837400_bib13","doi-asserted-by":"publisher","first-page":"1220","DOI":"10.1016\/j.cell.2010.05.006","article-title":"A large-scale conformational change couples membrane recruitment to cargo binding in the AP2 clathrin adaptor complex","volume":"141","author":"Jackson","year":"2010","journal-title":"Cell."},{"key":"2023072309062837400_bib14","doi-asserted-by":"publisher","first-page":"e00829","DOI":"10.7554\/eLife.00829","article-title":"An unmet actin requirement explains the mitotic inhibition of clathrin-mediated endocytosis","volume":"3","author":"Kaur","year":"2014","journal-title":"eLife."},{"key":"2023072309062837400_bib15","doi-asserted-by":"publisher","first-page":"976","DOI":"10.1038\/nature07422","article-title":"A structural explanation for the binding of endocytic dileucine motifs by the AP2 complex","volume":"456","author":"Kelly","year":"2008","journal-title":"Nature."},{"key":"2023072309062837400_bib16","doi-asserted-by":"publisher","first-page":"459","DOI":"10.1126\/science.1254836","article-title":"Clathrin adaptors. AP2 controls clathrin polymerization with a membrane-activated switch","volume":"345","author":"Kelly","year":"2014","journal-title":"Science."},{"key":"2023072309062837400_bib17","doi-asserted-by":"publisher","first-page":"5309","DOI":"10.1091\/mbc.E08-07-0712","article-title":"The AP-2 adaptor beta2 appendage scaffolds alternate cargo endocytosis","volume":"19","author":"Keyel","year":"2008","journal-title":"Mol. Biol. Cell."},{"key":"2023072309062837400_bib18","doi-asserted-by":"publisher","first-page":"a016725","DOI":"10.1101\/cshperspect.a016725","article-title":"Molecular structure, function, and dynamics of clathrin-mediated membrane traffic","volume":"6","author":"Kirchhausen","year":"2014","journal-title":"Cold Spring Harb. Perspect. Biol."},{"key":"2023072309062837400_bib19","doi-asserted-by":"publisher","first-page":"843","DOI":"10.1111\/j.1600-0854.2010.01056.x","article-title":"A screen for endocytic motifs","volume":"11","author":"Kozik","year":"2010","journal-title":"Traffic."},{"key":"2023072309062837400_bib20","doi-asserted-by":"publisher","first-page":"3070","DOI":"10.1091\/mbc.E14-06-1112","article-title":"Dual single-scission event analysis of constitutive transferrin receptor (TfR) endocytosis and ligand-triggered \u03b22-adrenergic receptor (\u03b22AR) or Mu-opioid receptor (MOR) endocytosis","volume":"25","author":"Lampe","year":"2014","journal-title":"Mol. Biol. Cell."},{"key":"2023072309062837400_bib21","doi-asserted-by":"publisher","first-page":"e57","DOI":"10.1371\/journal.pbio.1000057","article-title":"Cargo and dynamin regulate clathrin-coated pit maturation","volume":"7","author":"Loerke","year":"2009","journal-title":"PLoS Biol."},{"key":"2023072309062837400_bib22","doi-asserted-by":"publisher","first-page":"3251","DOI":"10.1091\/mbc.E09-03-0256","article-title":"Endocytic accessory proteins are functionally distinguished by their differential effects on the maturation of clathrin-coated pits","volume":"20","author":"Mettlen","year":"2009","journal-title":"Mol. Biol. Cell."},{"key":"2023072309062837400_bib23","doi-asserted-by":"publisher","first-page":"909","DOI":"10.1083\/jcb.200305145","article-title":"Clathrin-mediated endocytosis in AP-2-depleted cells","volume":"162","author":"Motley","year":"2003","journal-title":"J. Cell Biol."},{"key":"2023072309062837400_bib24","doi-asserted-by":"crossref","first-page":"10850","DOI":"10.1016\/S0021-9258(19)50096-5","article-title":"Recognition sites for clathrin-associated proteins AP-2 and AP-3 on clathrin triskelia","volume":"267","author":"Murphy","year":"1992","journal-title":"J. Biol. Chem."},{"key":"2023072309062837400_bib25","doi-asserted-by":"publisher","first-page":"4216","DOI":"10.1093\/emboj\/19.16.4216","article-title":"The structure and function of the beta 2-adaptin appendage domain","volume":"19","author":"Owen","year":"2000","journal-title":"EMBO J."},{"key":"2023072309062837400_bib26","doi-asserted-by":"publisher","first-page":"327","DOI":"10.1083\/jcb.200107140","article-title":"Assembly and function of AP-3 complexes in cells expressing mutant subunits","volume":"156","author":"Peden","year":"2002","journal-title":"J. Cell Biol."},{"key":"2023072309062837400_bib27","doi-asserted-by":"publisher","first-page":"988","DOI":"10.1016\/j.cell.2007.09.006","article-title":"Endocytosis of neurotransmitter receptors: location matters","volume":"130","author":"Puthenveedu","year":"2007","journal-title":"Cell."},{"key":"2023072309062837400_bib28","doi-asserted-by":"publisher","first-page":"40","DOI":"10.1111\/tra.12343","article-title":"Endocytosis of ligand-activated sphingosine 1-phosphate receptor 1 mediated by the clathrin-pathway","volume":"17","author":"Reeves","year":"2016","journal-title":"Traffic."},{"key":"2023072309062837400_bib29","doi-asserted-by":"publisher","first-page":"788","DOI":"10.1002\/embj.201386357","article-title":"Synaptic vesicle recycling: steps and principles","volume":"33","author":"Rizzoli","year":"2014","journal-title":"EMBO J."},{"key":"2023072309062837400_bib30","doi-asserted-by":"publisher","first-page":"1210","DOI":"10.1111\/tra.12335","article-title":"Forty years of clathrin-coated vesicles","volume":"16","author":"Robinson","year":"2015","journal-title":"Traffic."},{"key":"2023072309062837400_bib31","doi-asserted-by":"publisher","first-page":"31083","DOI":"10.1074\/jbc.270.52.31083","article-title":"A clathrin-binding site in the hinge of the beta 2 chain of mammalian AP-2 complexes","volume":"270","author":"Shih","year":"1995","journal-title":"J. Biol. Chem."},{"key":"2023072309062837400_bib32","doi-asserted-by":"publisher","first-page":"379","DOI":"10.1038\/nmeth.1904","article-title":"TULIPs: tunable, light-controlled interacting protein tags for cell biology","volume":"9","author":"Strickland","year":"2012","journal-title":"Nat. Methods."},{"key":"2023072309062837400_bib33","doi-asserted-by":"publisher","first-page":"21097","DOI":"10.1073\/pnas.0909204106","article-title":"Photoactivation mechanism of PAmCherry based on crystal structures of the protein in the dark and fluorescent states","volume":"106","author":"Subach","year":"2009","journal-title":"Proc. Natl. Acad. Sci. USA."},{"key":"2023072309062837400_bib34","doi-asserted-by":"publisher","first-page":"e1000604","DOI":"10.1371\/journal.pbio.1000604","article-title":"A high precision survey of the molecular dynamics of mammalian clathrin-mediated endocytosis","volume":"9","author":"Taylor","year":"2011","journal-title":"PLoS Biol."},{"key":"2023072309062837400_bib35","doi-asserted-by":"publisher","first-page":"1096","DOI":"10.1073\/pnas.97.3.1096","article-title":"Peptide-in-groove interactions link target proteins to the beta-propeller of clathrin","volume":"97","author":"ter Haar","year":"2000","journal-title":"Proc. Natl. Acad. Sci. USA."},{"key":"2023072309062837400_bib36","doi-asserted-by":"publisher","first-page":"583","DOI":"10.1038\/nrm2751","article-title":"Tickets to ride: selecting cargo for clathrin-regulated internalization","volume":"10","author":"Traub","year":"2009","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"2023072309062837400_bib37","doi-asserted-by":"publisher","first-page":"111","DOI":"10.1038\/nature14128","article-title":"Optogenetic control of organelle transport and positioning","volume":"518","author":"van Bergeijk","year":"2015","journal-title":"Nature."},{"key":"2023072309062837400_bib38","doi-asserted-by":"publisher","first-page":"641","DOI":"10.1083\/jcb.201603025","article-title":"Local RhoA activation induces cytokinetic furrows independent of spindle position and cell cycle stage","volume":"213","author":"Wagner","year":"2016","journal-title":"J. Cell Biol."},{"key":"2023072309062837400_bib39","doi-asserted-by":"publisher","first-page":"70","DOI":"10.1111\/j.1600-0854.2011.01289.x","article-title":"Functional analysis of interaction sites on the N-terminal domain of clathrin heavy chain","volume":"13","author":"Willox","year":"2012","journal-title":"Traffic."},{"key":"2023072309062837400_bib40","doi-asserted-by":"publisher","first-page":"2378","DOI":"10.1091\/mbc.E12-12-0860","article-title":"Chemical-genetic disruption of clathrin function spares adaptor complex 3-dependent endosome vesicle biogenesis","volume":"24","author":"Zlatic","year":"2013","journal-title":"Mol. Biol. Cell."}],"container-title":["Journal of Cell Biology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/rupress.org\/jcb\/article-pdf\/216\/12\/4351\/1597310\/jcb_201702188.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/rupress.org\/jcb\/article-pdf\/216\/12\/4351\/1597310\/jcb_201702188.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,7,23]],"date-time":"2023-07-23T05:09:34Z","timestamp":1690088974000},"score":1,"resource":{"primary":{"URL":"https:\/\/rupress.org\/jcb\/article\/216\/12\/4351\/38841\/New-tools-for-hot-wiring-clathrin-mediated"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2017,9,27]]},"references-count":40,"journal-issue":{"issue":"12","published-print":{"date-parts":[[2017,12,4]]}},"URL":"https:\/\/doi.org\/10.1083\/jcb.201702188","relation":{"has-preprint":[{"id-type":"doi","id":"10.1101\/061986","asserted-by":"object"}]},"ISSN":["0021-9525","1540-8140"],"issn-type":[{"value":"0021-9525","type":"print"},{"value":"1540-8140","type":"electronic"}],"subject":[],"published":{"date-parts":[[2017,9,27]]}}}