Molecularly Distinct Clathrin-Coated Pits Differentially Impact EGFR Fate and Signaling
Summary: Adaptor protein 2 (AP2) is a major constituent of clathrin-coated pits (CCPs). Whether it is essential for all forms of clathrin-mediated endocytosis (CME) in mammalian cells is an open issue. Here, we demonstrate, by live TIRF microscopy, the existence of a subclass of relatively short-liv...
| Main Authors: | , , , , , , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2019-06-01
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| Series: | Cell Reports |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S221112471930628X |
| _version_ | 1818502908468002816 |
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| author | Roberta Pascolutti Veronica Algisi Alexia Conte Andrea Raimondi Mithun Pasham Srigokul Upadhyayula Raphael Gaudin Tanja Maritzen Elisa Barbieri Giusi Caldieri Chiara Tordonato Stefano Confalonieri Stefano Freddi Maria Grazia Malabarba Elena Maspero Simona Polo Carlo Tacchetti Volker Haucke Tom Kirchhausen Pier Paolo Di Fiore Sara Sigismund |
| author_facet | Roberta Pascolutti Veronica Algisi Alexia Conte Andrea Raimondi Mithun Pasham Srigokul Upadhyayula Raphael Gaudin Tanja Maritzen Elisa Barbieri Giusi Caldieri Chiara Tordonato Stefano Confalonieri Stefano Freddi Maria Grazia Malabarba Elena Maspero Simona Polo Carlo Tacchetti Volker Haucke Tom Kirchhausen Pier Paolo Di Fiore Sara Sigismund |
| author_sort | Roberta Pascolutti |
| collection | DOAJ |
| description | Summary: Adaptor protein 2 (AP2) is a major constituent of clathrin-coated pits (CCPs). Whether it is essential for all forms of clathrin-mediated endocytosis (CME) in mammalian cells is an open issue. Here, we demonstrate, by live TIRF microscopy, the existence of a subclass of relatively short-lived CCPs lacking AP2 under physiological, unperturbed conditions. This subclass is retained in AP2-knockout cells and is able to support the internalization of epidermal growth factor receptor (EGFR) but not of transferrin receptor (TfR). The AP2-independent internalization mechanism relies on the endocytic adaptors eps15, eps15L1, and epsin1. The absence of AP2 impairs the recycling of the EGFR to the cell surface, thereby augmenting its degradation. Accordingly, under conditions of AP2 ablation, we detected dampening of EGFR-dependent AKT signaling and cell migration, arguing that distinct classes of CCPs could provide specialized functions in regulating EGFR recycling and signaling. : EGFR signaling controls different cell physiological processes, including proliferation and migration. Pascolutti et al. describe an additional layer of regulation of EGFR signaling, relying on the sequestration of receptors into molecularly distinct clathrin-coated vesicles that regulate receptor fate toward recycling versus degradation, with impact on the final cellular output. Keywords: EGFR, endocytosis, clathrin-coated pits, AP2, eps15, epsin, endocytic adaptors, signaling, receptor degradation, recycling, transcription |
| first_indexed | 2024-12-10T21:16:41Z |
| format | Article |
| id | doaj.art-621ad877eaa14894b7e9f36e39fad5bd |
| institution | Directory Open Access Journal |
| issn | 2211-1247 |
| language | English |
| last_indexed | 2024-12-10T21:16:41Z |
| publishDate | 2019-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Cell Reports |
| spelling | doaj.art-621ad877eaa14894b7e9f36e39fad5bd2022-12-22T01:33:17ZengElsevierCell Reports2211-12472019-06-01271030493061.e6Molecularly Distinct Clathrin-Coated Pits Differentially Impact EGFR Fate and SignalingRoberta Pascolutti0Veronica Algisi1Alexia Conte2Andrea Raimondi3Mithun Pasham4Srigokul Upadhyayula5Raphael Gaudin6Tanja Maritzen7Elisa Barbieri8Giusi Caldieri9Chiara Tordonato10Stefano Confalonieri11Stefano Freddi12Maria Grazia Malabarba13Elena Maspero14Simona Polo15Carlo Tacchetti16Volker Haucke17Tom Kirchhausen18Pier Paolo Di Fiore19Sara Sigismund20IFOM, Fondazione Istituto FIRC di Oncologia Molecolare, Via Adamello 16, 20139 Milan, ItalyIFOM, Fondazione Istituto FIRC di Oncologia Molecolare, Via Adamello 16, 20139 Milan, ItalyIFOM, Fondazione Istituto FIRC di Oncologia Molecolare, Via Adamello 16, 20139 Milan, Italy; Istituto Europeo di Oncologia IRCCS, Via Ripamonti 435, 20141 Milan, ItalyExperimental Imaging Centre, Istituto di Ricovero e Cura a Carattere Scientifico (IRCCS), San Raffaele Scientific Institute, via Olgettina 58, 20132 Milan, ItalyProgram in Cellular and Molecular Medicine, Boston Children’s Hospital, Boston, MA 02115, USAProgram in Cellular and Molecular Medicine, Boston Children’s Hospital, Boston, MA 02115, USA; Department of Pediatrics, Harvard Medical School, Boston, MA 02115, USA; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20147, USAProgram in Cellular and Molecular Medicine, Boston Children’s Hospital, Boston, MA 02115, USA; Department of Cell Biology, Harvard Medical School, Boston, MA 02115, USA; Institut de Recherche en Infectiologie de Montpellier, UMR 9004, CNRS/UM, 1919 route de Mende, 34293 Montpellier cedex 5, FranceLeibniz-Forschungsinstitut für Molekulare Pharmakologie (FMP), Robert-Roessle-Straße 10, 13125 Berlin, GermanyIFOM, Fondazione Istituto FIRC di Oncologia Molecolare, Via Adamello 16, 20139 Milan, Italy; Istituto Europeo di Oncologia IRCCS, Via Ripamonti 435, 20141 Milan, ItalyIFOM, Fondazione Istituto FIRC di Oncologia Molecolare, Via Adamello 16, 20139 Milan, Italy; Istituto Europeo di Oncologia IRCCS, Via Ripamonti 435, 20141 Milan, Italy; Università degli Studi di Milano, Dipartimento di Oncologia ed Emato-oncologia, Via Santa Sofia 9/1, 20122 Milan, ItalyIstituto Europeo di Oncologia IRCCS, Via Ripamonti 435, 20141 Milan, ItalyIFOM, Fondazione Istituto FIRC di Oncologia Molecolare, Via Adamello 16, 20139 Milan, Italy; Istituto Europeo di Oncologia IRCCS, Via Ripamonti 435, 20141 Milan, ItalyIstituto Europeo di Oncologia IRCCS, Via Ripamonti 435, 20141 Milan, ItalyIFOM, Fondazione Istituto FIRC di Oncologia Molecolare, Via Adamello 16, 20139 Milan, Italy; Istituto Europeo di Oncologia IRCCS, Via Ripamonti 435, 20141 Milan, Italy; Università degli Studi di Milano, Dipartimento di Oncologia ed Emato-oncologia, Via Santa Sofia 9/1, 20122 Milan, ItalyIFOM, Fondazione Istituto FIRC di Oncologia Molecolare, Via Adamello 16, 20139 Milan, ItalyIFOM, Fondazione Istituto FIRC di Oncologia Molecolare, Via Adamello 16, 20139 Milan, Italy; Università degli Studi di Milano, Dipartimento di Oncologia ed Emato-oncologia, Via Santa Sofia 9/1, 20122 Milan, ItalyExperimental Imaging Centre, Istituto di Ricovero e Cura a Carattere Scientifico (IRCCS), San Raffaele Scientific Institute, via Olgettina 58, 20132 Milan, Italy; Università Vita-Salute San Raffaele, Milan, ItalyLeibniz-Forschungsinstitut für Molekulare Pharmakologie (FMP), Robert-Roessle-Straße 10, 13125 Berlin, GermanyProgram in Cellular and Molecular Medicine, Boston Children’s Hospital, Boston, MA 02115, USA; Department of Pediatrics, Harvard Medical School, Boston, MA 02115, USA; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20147, USA; Department of Cell Biology, Harvard Medical School, Boston, MA 02115, USAIFOM, Fondazione Istituto FIRC di Oncologia Molecolare, Via Adamello 16, 20139 Milan, Italy; Istituto Europeo di Oncologia IRCCS, Via Ripamonti 435, 20141 Milan, Italy; Università degli Studi di Milano, Dipartimento di Oncologia ed Emato-oncologia, Via Santa Sofia 9/1, 20122 Milan, ItalyIFOM, Fondazione Istituto FIRC di Oncologia Molecolare, Via Adamello 16, 20139 Milan, Italy; Istituto Europeo di Oncologia IRCCS, Via Ripamonti 435, 20141 Milan, Italy; Università degli Studi di Milano, Dipartimento di Oncologia ed Emato-oncologia, Via Santa Sofia 9/1, 20122 Milan, Italy; Corresponding authorSummary: Adaptor protein 2 (AP2) is a major constituent of clathrin-coated pits (CCPs). Whether it is essential for all forms of clathrin-mediated endocytosis (CME) in mammalian cells is an open issue. Here, we demonstrate, by live TIRF microscopy, the existence of a subclass of relatively short-lived CCPs lacking AP2 under physiological, unperturbed conditions. This subclass is retained in AP2-knockout cells and is able to support the internalization of epidermal growth factor receptor (EGFR) but not of transferrin receptor (TfR). The AP2-independent internalization mechanism relies on the endocytic adaptors eps15, eps15L1, and epsin1. The absence of AP2 impairs the recycling of the EGFR to the cell surface, thereby augmenting its degradation. Accordingly, under conditions of AP2 ablation, we detected dampening of EGFR-dependent AKT signaling and cell migration, arguing that distinct classes of CCPs could provide specialized functions in regulating EGFR recycling and signaling. : EGFR signaling controls different cell physiological processes, including proliferation and migration. Pascolutti et al. describe an additional layer of regulation of EGFR signaling, relying on the sequestration of receptors into molecularly distinct clathrin-coated vesicles that regulate receptor fate toward recycling versus degradation, with impact on the final cellular output. Keywords: EGFR, endocytosis, clathrin-coated pits, AP2, eps15, epsin, endocytic adaptors, signaling, receptor degradation, recycling, transcriptionhttp://www.sciencedirect.com/science/article/pii/S221112471930628X |
| spellingShingle | Roberta Pascolutti Veronica Algisi Alexia Conte Andrea Raimondi Mithun Pasham Srigokul Upadhyayula Raphael Gaudin Tanja Maritzen Elisa Barbieri Giusi Caldieri Chiara Tordonato Stefano Confalonieri Stefano Freddi Maria Grazia Malabarba Elena Maspero Simona Polo Carlo Tacchetti Volker Haucke Tom Kirchhausen Pier Paolo Di Fiore Sara Sigismund Molecularly Distinct Clathrin-Coated Pits Differentially Impact EGFR Fate and Signaling Cell Reports |
| title | Molecularly Distinct Clathrin-Coated Pits Differentially Impact EGFR Fate and Signaling |
| title_full | Molecularly Distinct Clathrin-Coated Pits Differentially Impact EGFR Fate and Signaling |
| title_fullStr | Molecularly Distinct Clathrin-Coated Pits Differentially Impact EGFR Fate and Signaling |
| title_full_unstemmed | Molecularly Distinct Clathrin-Coated Pits Differentially Impact EGFR Fate and Signaling |
| title_short | Molecularly Distinct Clathrin-Coated Pits Differentially Impact EGFR Fate and Signaling |
| title_sort | molecularly distinct clathrin coated pits differentially impact egfr fate and signaling |
| url | http://www.sciencedirect.com/science/article/pii/S221112471930628X |
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