Clathrin-Mediated Endocytosis Regulates a Balance between Opposing Signals to Maintain the Pluripotent State of Embryonic Stem Cells
Summary: Endocytosis is implicated in the maintenance of embryonic stem cell (ESC) pluripotency, although its exact role and the identity of molecular players remain poorly understood. Here, we show that the clathrin heavy chain (CLTC), involved in clathrin-mediated endocytosis (CME), is vital for m...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2019-01-01
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| Series: | Stem Cell Reports |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2213671118304879 |
| _version_ | 1818603295254511616 |
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| author | Yadavalli V. Narayana Chetan Gadgil Ridim D. Mote Raghav Rajan Deepa Subramanyam |
| author_facet | Yadavalli V. Narayana Chetan Gadgil Ridim D. Mote Raghav Rajan Deepa Subramanyam |
| author_sort | Yadavalli V. Narayana |
| collection | DOAJ |
| description | Summary: Endocytosis is implicated in the maintenance of embryonic stem cell (ESC) pluripotency, although its exact role and the identity of molecular players remain poorly understood. Here, we show that the clathrin heavy chain (CLTC), involved in clathrin-mediated endocytosis (CME), is vital for maintaining mouse ESC (mESC) pluripotency. Knockdown of Cltc resulted in a loss of pluripotency accompanied by reduced E-cadherin (E-CAD) levels and increased levels of transforming growth factor β (TGF-β) and extracellular signal-regulated kinase (ERK) signaling. We demonstrate that both E-CAD and TGF-β receptor type 1 (TGF-βR1) are internalized through CME in mESCs. While E-CAD is recycled, TGF-βR1 is targeted for lysosomal degradation thus maintaining inverse levels of these molecules. Finally, we show that E-CAD interacts with ERK, and that the decreased pluripotency upon CME loss can be rescued by inhibiting TGF-βR, MEK, and GSK3β, or overexpressing E-CAD. Our results demonstrate that CME is critical for balancing signaling outputs to regulate ESC pluripotency, and possibly cell fate choices in early development. : Subramanyam and colleagues demonstrate that clathrin-mediated endocytosis (CME) is necessary to maintain ESC pluripotency by regulating the trafficking of E-CAD and TGF-βR1. Loss of CME results in a loss of pluripotency characterized by decreased E-CAD and enhanced TGF-βR1 and MEK signaling. Expression of pluripotency markers in ESCs lacking Cltc can be restored by inhibiting TGF-βR1 or MEK or by overexpressing E-CAD. Keywords: embryonic stem cells, pluripotency, trafficking, E-cadherin, clathrin, recycling, TGF-β, epithelial |
| first_indexed | 2024-12-16T13:20:54Z |
| format | Article |
| id | doaj.art-f4126a6c03f648f0b20955a325543c6e |
| institution | Directory Open Access Journal |
| issn | 2213-6711 |
| language | English |
| last_indexed | 2024-12-16T13:20:54Z |
| publishDate | 2019-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Stem Cell Reports |
| spelling | doaj.art-f4126a6c03f648f0b20955a325543c6e2022-12-21T22:30:21ZengElsevierStem Cell Reports2213-67112019-01-01121152164Clathrin-Mediated Endocytosis Regulates a Balance between Opposing Signals to Maintain the Pluripotent State of Embryonic Stem CellsYadavalli V. Narayana0Chetan Gadgil1Ridim D. Mote2Raghav Rajan3Deepa Subramanyam4National Centre for Cell Science, SP Pune University, Ganeshkhind, Pune 411007, India; Savitribai Phule Pune University, Ganeshkhind, Pune 411007, IndiaChemical Engineering Department, National Chemical Laboratory, Dr. Homi Bhabha Road, Pune 411008, IndiaNational Centre for Cell Science, SP Pune University, Ganeshkhind, Pune 411007, IndiaIndian Institute of Science Education and Research, Dr. Homi Bhabha Road, NCL Colony, Pune 411008, IndiaNational Centre for Cell Science, SP Pune University, Ganeshkhind, Pune 411007, India; Corresponding authorSummary: Endocytosis is implicated in the maintenance of embryonic stem cell (ESC) pluripotency, although its exact role and the identity of molecular players remain poorly understood. Here, we show that the clathrin heavy chain (CLTC), involved in clathrin-mediated endocytosis (CME), is vital for maintaining mouse ESC (mESC) pluripotency. Knockdown of Cltc resulted in a loss of pluripotency accompanied by reduced E-cadherin (E-CAD) levels and increased levels of transforming growth factor β (TGF-β) and extracellular signal-regulated kinase (ERK) signaling. We demonstrate that both E-CAD and TGF-β receptor type 1 (TGF-βR1) are internalized through CME in mESCs. While E-CAD is recycled, TGF-βR1 is targeted for lysosomal degradation thus maintaining inverse levels of these molecules. Finally, we show that E-CAD interacts with ERK, and that the decreased pluripotency upon CME loss can be rescued by inhibiting TGF-βR, MEK, and GSK3β, or overexpressing E-CAD. Our results demonstrate that CME is critical for balancing signaling outputs to regulate ESC pluripotency, and possibly cell fate choices in early development. : Subramanyam and colleagues demonstrate that clathrin-mediated endocytosis (CME) is necessary to maintain ESC pluripotency by regulating the trafficking of E-CAD and TGF-βR1. Loss of CME results in a loss of pluripotency characterized by decreased E-CAD and enhanced TGF-βR1 and MEK signaling. Expression of pluripotency markers in ESCs lacking Cltc can be restored by inhibiting TGF-βR1 or MEK or by overexpressing E-CAD. Keywords: embryonic stem cells, pluripotency, trafficking, E-cadherin, clathrin, recycling, TGF-β, epithelialhttp://www.sciencedirect.com/science/article/pii/S2213671118304879 |
| spellingShingle | Yadavalli V. Narayana Chetan Gadgil Ridim D. Mote Raghav Rajan Deepa Subramanyam Clathrin-Mediated Endocytosis Regulates a Balance between Opposing Signals to Maintain the Pluripotent State of Embryonic Stem Cells Stem Cell Reports |
| title | Clathrin-Mediated Endocytosis Regulates a Balance between Opposing Signals to Maintain the Pluripotent State of Embryonic Stem Cells |
| title_full | Clathrin-Mediated Endocytosis Regulates a Balance between Opposing Signals to Maintain the Pluripotent State of Embryonic Stem Cells |
| title_fullStr | Clathrin-Mediated Endocytosis Regulates a Balance between Opposing Signals to Maintain the Pluripotent State of Embryonic Stem Cells |
| title_full_unstemmed | Clathrin-Mediated Endocytosis Regulates a Balance between Opposing Signals to Maintain the Pluripotent State of Embryonic Stem Cells |
| title_short | Clathrin-Mediated Endocytosis Regulates a Balance between Opposing Signals to Maintain the Pluripotent State of Embryonic Stem Cells |
| title_sort | clathrin mediated endocytosis regulates a balance between opposing signals to maintain the pluripotent state of embryonic stem cells |
| url | http://www.sciencedirect.com/science/article/pii/S2213671118304879 |
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