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Retromer Binding to FAM21 and the WASH Complex Is Perturbed by the Parkinson Disease-Linked VPS35(D620N) Mutation
Retromer is a protein assembly that plays a central role in orchestrating export of transmembrane-spanning cargo proteins from endosomes into retrieval pathways destined for the Golgi apparatus and the plasma membrane [1]. Recently, a specific mutation in the retromer component VPS35, VPS35(D620N),...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Cell Press
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4110399/ https://www.ncbi.nlm.nih.gov/pubmed/24980502 http://dx.doi.org/10.1016/j.cub.2014.06.024 |
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author | McGough, Ian J. Steinberg, Florian Jia, Da Barbuti, Peter A. McMillan, Kirsty J. Heesom, Kate J. Whone, Alan L. Caldwell, Maeve A. Billadeau, Daniel D. Rosen, Michael K. Cullen, Peter J. |
author_facet | McGough, Ian J. Steinberg, Florian Jia, Da Barbuti, Peter A. McMillan, Kirsty J. Heesom, Kate J. Whone, Alan L. Caldwell, Maeve A. Billadeau, Daniel D. Rosen, Michael K. Cullen, Peter J. |
author_sort | McGough, Ian J. |
collection | PubMed |
description | Retromer is a protein assembly that plays a central role in orchestrating export of transmembrane-spanning cargo proteins from endosomes into retrieval pathways destined for the Golgi apparatus and the plasma membrane [1]. Recently, a specific mutation in the retromer component VPS35, VPS35(D620N), has linked retromer dysfunction to familial autosomal dominant and sporadic Parkinson disease [2, 3]. However, the effect of this mutation on retromer function remains poorly characterized. Here we established that in cells expressing VPS35(D620N) there is a perturbation in endosome-to-TGN transport but not endosome-to-plasma membrane recycling, which we confirm in patient cells harboring the VPS35(D620N) mutation. Through comparative stable isotope labeling by amino acids in cell culture (SILAC)-based analysis of wild-type VPS35 versus the VPS35(D620N) mutant interactomes, we establish that the major defect of the D620N mutation lies in the association to the actin-nucleating Wiskott-Aldrich syndrome and SCAR homolog (WASH) complex. Moreover, using isothermal calorimetry, we establish that the primary defect of the VPS35(D620N) mutant is a 2.2 ± 0.5-fold decrease in affinity for the WASH complex component FAM21. These data define the primary molecular defect in retromer assembly that arises from the VPS35(D620N) mutation and, by revealing functional effects on retromer-mediated endosome-to-TGN transport, provide new insight into retromer deregulation in Parkinson disease. |
format | Online Article Text |
id | pubmed-4110399 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Cell Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-41103992014-07-25 Retromer Binding to FAM21 and the WASH Complex Is Perturbed by the Parkinson Disease-Linked VPS35(D620N) Mutation McGough, Ian J. Steinberg, Florian Jia, Da Barbuti, Peter A. McMillan, Kirsty J. Heesom, Kate J. Whone, Alan L. Caldwell, Maeve A. Billadeau, Daniel D. Rosen, Michael K. Cullen, Peter J. Curr Biol Report Retromer is a protein assembly that plays a central role in orchestrating export of transmembrane-spanning cargo proteins from endosomes into retrieval pathways destined for the Golgi apparatus and the plasma membrane [1]. Recently, a specific mutation in the retromer component VPS35, VPS35(D620N), has linked retromer dysfunction to familial autosomal dominant and sporadic Parkinson disease [2, 3]. However, the effect of this mutation on retromer function remains poorly characterized. Here we established that in cells expressing VPS35(D620N) there is a perturbation in endosome-to-TGN transport but not endosome-to-plasma membrane recycling, which we confirm in patient cells harboring the VPS35(D620N) mutation. Through comparative stable isotope labeling by amino acids in cell culture (SILAC)-based analysis of wild-type VPS35 versus the VPS35(D620N) mutant interactomes, we establish that the major defect of the D620N mutation lies in the association to the actin-nucleating Wiskott-Aldrich syndrome and SCAR homolog (WASH) complex. Moreover, using isothermal calorimetry, we establish that the primary defect of the VPS35(D620N) mutant is a 2.2 ± 0.5-fold decrease in affinity for the WASH complex component FAM21. These data define the primary molecular defect in retromer assembly that arises from the VPS35(D620N) mutation and, by revealing functional effects on retromer-mediated endosome-to-TGN transport, provide new insight into retromer deregulation in Parkinson disease. Cell Press 2014-07-21 /pmc/articles/PMC4110399/ /pubmed/24980502 http://dx.doi.org/10.1016/j.cub.2014.06.024 Text en © 2014 The Authors http://creativecommons.org/licenses/by/3.0/ This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/3.0/). |
spellingShingle | Report McGough, Ian J. Steinberg, Florian Jia, Da Barbuti, Peter A. McMillan, Kirsty J. Heesom, Kate J. Whone, Alan L. Caldwell, Maeve A. Billadeau, Daniel D. Rosen, Michael K. Cullen, Peter J. Retromer Binding to FAM21 and the WASH Complex Is Perturbed by the Parkinson Disease-Linked VPS35(D620N) Mutation |
title | Retromer Binding to FAM21 and the WASH Complex Is Perturbed by the Parkinson Disease-Linked VPS35(D620N) Mutation |
title_full | Retromer Binding to FAM21 and the WASH Complex Is Perturbed by the Parkinson Disease-Linked VPS35(D620N) Mutation |
title_fullStr | Retromer Binding to FAM21 and the WASH Complex Is Perturbed by the Parkinson Disease-Linked VPS35(D620N) Mutation |
title_full_unstemmed | Retromer Binding to FAM21 and the WASH Complex Is Perturbed by the Parkinson Disease-Linked VPS35(D620N) Mutation |
title_short | Retromer Binding to FAM21 and the WASH Complex Is Perturbed by the Parkinson Disease-Linked VPS35(D620N) Mutation |
title_sort | retromer binding to fam21 and the wash complex is perturbed by the parkinson disease-linked vps35(d620n) mutation |
topic | Report |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4110399/ https://www.ncbi.nlm.nih.gov/pubmed/24980502 http://dx.doi.org/10.1016/j.cub.2014.06.024 |
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