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Interaction of phosphorylated Rab11-FIP2 with Eps15 regulates apical junction composition
MARK2 regulates the establishment of polarity in Madin–Darby canine kidney (MDCK) cells in part through phosphorylation of serine 227 of Rab11-FIP2. We identified Eps15 as an interacting partner of phospho-S227-Rab11-FIP2 (pS227-FIP2). During recovery from low calcium, Eps15 localized to the lateral...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The American Society for Cell Biology
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5391185/ https://www.ncbi.nlm.nih.gov/pubmed/28228550 http://dx.doi.org/10.1091/mbc.E16-04-0214 |
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author | Lapierre, Lynne A. Manning, Elizabeth H. Mitchell, Kenya M. Caldwell, Cathy M. Goldenring, James R. |
author_facet | Lapierre, Lynne A. Manning, Elizabeth H. Mitchell, Kenya M. Caldwell, Cathy M. Goldenring, James R. |
author_sort | Lapierre, Lynne A. |
collection | PubMed |
description | MARK2 regulates the establishment of polarity in Madin–Darby canine kidney (MDCK) cells in part through phosphorylation of serine 227 of Rab11-FIP2. We identified Eps15 as an interacting partner of phospho-S227-Rab11-FIP2 (pS227-FIP2). During recovery from low calcium, Eps15 localized to the lateral membrane before pS227-FIP2 arrival. Later in recovery, Eps15 and pS227-FIP2 colocalized at the lateral membrane. In MDCK cells expressing the pseudophosphorylated FIP2 mutant FIP2(S227E), during recovery from low calcium, Eps15 was trapped and never localized to the lateral membrane. Mutation of any of the three NPF domains within GFP-FIP2(S227E) rescued Eps15 localization at the lateral membrane and reestablished single-lumen cyst formation in GFP-FIP2(S227E)–expressing cells in three-dimensional (3D) culture. Whereas expression of GFP-FIP2(S227E) induced the loss of E-cadherin and occludin, mutation of any of the NPF domains of GFP-FIP2(S227E) reestablished both proteins at the apical junctions. Knockdown of Eps15 altered the spatial and temporal localization of pS227-FIP2 and also elicited formation of multiple lumens in MDCK 3D cysts. Thus an interaction of Eps15 and pS227-FIP2 at the appropriate time and location in polarizing cells is necessary for proper establishment of epithelial polarity. |
format | Online Article Text |
id | pubmed-5391185 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | The American Society for Cell Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-53911852017-06-30 Interaction of phosphorylated Rab11-FIP2 with Eps15 regulates apical junction composition Lapierre, Lynne A. Manning, Elizabeth H. Mitchell, Kenya M. Caldwell, Cathy M. Goldenring, James R. Mol Biol Cell Articles MARK2 regulates the establishment of polarity in Madin–Darby canine kidney (MDCK) cells in part through phosphorylation of serine 227 of Rab11-FIP2. We identified Eps15 as an interacting partner of phospho-S227-Rab11-FIP2 (pS227-FIP2). During recovery from low calcium, Eps15 localized to the lateral membrane before pS227-FIP2 arrival. Later in recovery, Eps15 and pS227-FIP2 colocalized at the lateral membrane. In MDCK cells expressing the pseudophosphorylated FIP2 mutant FIP2(S227E), during recovery from low calcium, Eps15 was trapped and never localized to the lateral membrane. Mutation of any of the three NPF domains within GFP-FIP2(S227E) rescued Eps15 localization at the lateral membrane and reestablished single-lumen cyst formation in GFP-FIP2(S227E)–expressing cells in three-dimensional (3D) culture. Whereas expression of GFP-FIP2(S227E) induced the loss of E-cadherin and occludin, mutation of any of the NPF domains of GFP-FIP2(S227E) reestablished both proteins at the apical junctions. Knockdown of Eps15 altered the spatial and temporal localization of pS227-FIP2 and also elicited formation of multiple lumens in MDCK 3D cysts. Thus an interaction of Eps15 and pS227-FIP2 at the appropriate time and location in polarizing cells is necessary for proper establishment of epithelial polarity. The American Society for Cell Biology 2017-04-15 /pmc/articles/PMC5391185/ /pubmed/28228550 http://dx.doi.org/10.1091/mbc.E16-04-0214 Text en © 2017 Lapierre et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0). “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society for Cell Biology. |
spellingShingle | Articles Lapierre, Lynne A. Manning, Elizabeth H. Mitchell, Kenya M. Caldwell, Cathy M. Goldenring, James R. Interaction of phosphorylated Rab11-FIP2 with Eps15 regulates apical junction composition |
title | Interaction of phosphorylated Rab11-FIP2 with Eps15 regulates apical junction composition |
title_full | Interaction of phosphorylated Rab11-FIP2 with Eps15 regulates apical junction composition |
title_fullStr | Interaction of phosphorylated Rab11-FIP2 with Eps15 regulates apical junction composition |
title_full_unstemmed | Interaction of phosphorylated Rab11-FIP2 with Eps15 regulates apical junction composition |
title_short | Interaction of phosphorylated Rab11-FIP2 with Eps15 regulates apical junction composition |
title_sort | interaction of phosphorylated rab11-fip2 with eps15 regulates apical junction composition |
topic | Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5391185/ https://www.ncbi.nlm.nih.gov/pubmed/28228550 http://dx.doi.org/10.1091/mbc.E16-04-0214 |
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