RNF13 Dileucine Motif Variants L311S and L312P Interfere with Endosomal Localization and AP-3 Complex Association
Developmental and epileptic encephalopathies (DEE) are rare and serious neurological disorders characterized by severe epilepsy with refractory seizures and a significant developmental delay. Recently, DEE73 was linked to genetic alterations of the RNF13 gene, which convert positions 311 or 312 in t...
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MDPI AG
2021-11-01
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| author | Valérie C. Cabana Antoine Y. Bouchard Audrey M. Sénécal Kim Ghilarducci Saïd Kourrich Laurent Cappadocia Marc P. Lussier |
| author_facet | Valérie C. Cabana Antoine Y. Bouchard Audrey M. Sénécal Kim Ghilarducci Saïd Kourrich Laurent Cappadocia Marc P. Lussier |
| author_sort | Valérie C. Cabana |
| collection | DOAJ |
| description | Developmental and epileptic encephalopathies (DEE) are rare and serious neurological disorders characterized by severe epilepsy with refractory seizures and a significant developmental delay. Recently, DEE73 was linked to genetic alterations of the RNF13 gene, which convert positions 311 or 312 in the RNF13 protein from leucine to serine or proline, respectively (L311S and L312P). Using a fluorescence microscopy approach to investigate the molecular and cellular mechanisms affected by RNF13 protein variants, the current study shows that wild-type RNF13 localizes extensively with endosomes and lysosomes, while L311S and L312P do not extensively colocalize with the lysosomal marker Lamp1. Our results show that RNF13 L311S and L312P proteins affect the size of endosomal vesicles along with the temporal and spatial progression of fluorescently labeled epidermal growth factor, but not transferrin, in the endolysosomal system. Furthermore, GST-pulldown and co-immunoprecipitation show that RNF13 variants disrupt association with AP-3 complex. Knockdown of AP-3 complex subunit AP3D1 alters the lysosomal localization of wild-type RNF13 and similarly affects the size of endosomal vesicles. Importantly, our study provides a first step toward understanding the cellular and molecular mechanism altered by DEE73-associated genetic variations of RNF13. |
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| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-10T05:36:02Z |
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| spelling | doaj.art-19041a9cefa4484d8aa32a40fe48c8682023-11-22T22:50:46ZengMDPI AGCells2073-44092021-11-011011306310.3390/cells10113063RNF13 Dileucine Motif Variants L311S and L312P Interfere with Endosomal Localization and AP-3 Complex AssociationValérie C. Cabana0Antoine Y. Bouchard1Audrey M. Sénécal2Kim Ghilarducci3Saïd Kourrich4Laurent Cappadocia5Marc P. Lussier6Département de Chimie, Université du Québec à Montréal, Montréal, QC H2X 2J6, CanadaDépartement de Chimie, Université du Québec à Montréal, Montréal, QC H2X 2J6, CanadaDépartement de Chimie, Université du Québec à Montréal, Montréal, QC H2X 2J6, CanadaDépartement de Chimie, Université du Québec à Montréal, Montréal, QC H2X 2J6, CanadaCentre d’Excellence en Recherche sur les Maladies Orphelines—Fondation Courtois (CERMO-FC), Université du Québec à Montréal, Montréal, QC H2X 3Y7, CanadaDépartement de Chimie, Université du Québec à Montréal, Montréal, QC H2X 2J6, CanadaDépartement de Chimie, Université du Québec à Montréal, Montréal, QC H2X 2J6, CanadaDevelopmental and epileptic encephalopathies (DEE) are rare and serious neurological disorders characterized by severe epilepsy with refractory seizures and a significant developmental delay. Recently, DEE73 was linked to genetic alterations of the RNF13 gene, which convert positions 311 or 312 in the RNF13 protein from leucine to serine or proline, respectively (L311S and L312P). Using a fluorescence microscopy approach to investigate the molecular and cellular mechanisms affected by RNF13 protein variants, the current study shows that wild-type RNF13 localizes extensively with endosomes and lysosomes, while L311S and L312P do not extensively colocalize with the lysosomal marker Lamp1. Our results show that RNF13 L311S and L312P proteins affect the size of endosomal vesicles along with the temporal and spatial progression of fluorescently labeled epidermal growth factor, but not transferrin, in the endolysosomal system. Furthermore, GST-pulldown and co-immunoprecipitation show that RNF13 variants disrupt association with AP-3 complex. Knockdown of AP-3 complex subunit AP3D1 alters the lysosomal localization of wild-type RNF13 and similarly affects the size of endosomal vesicles. Importantly, our study provides a first step toward understanding the cellular and molecular mechanism altered by DEE73-associated genetic variations of RNF13.https://www.mdpi.com/2073-4409/10/11/3063RNF13AP-3 complexendosomeslysosomesintracellular traffickinggenetic mutations |
| spellingShingle | Valérie C. Cabana Antoine Y. Bouchard Audrey M. Sénécal Kim Ghilarducci Saïd Kourrich Laurent Cappadocia Marc P. Lussier RNF13 Dileucine Motif Variants L311S and L312P Interfere with Endosomal Localization and AP-3 Complex Association Cells RNF13 AP-3 complex endosomes lysosomes intracellular trafficking genetic mutations |
| title | RNF13 Dileucine Motif Variants L311S and L312P Interfere with Endosomal Localization and AP-3 Complex Association |
| title_full | RNF13 Dileucine Motif Variants L311S and L312P Interfere with Endosomal Localization and AP-3 Complex Association |
| title_fullStr | RNF13 Dileucine Motif Variants L311S and L312P Interfere with Endosomal Localization and AP-3 Complex Association |
| title_full_unstemmed | RNF13 Dileucine Motif Variants L311S and L312P Interfere with Endosomal Localization and AP-3 Complex Association |
| title_short | RNF13 Dileucine Motif Variants L311S and L312P Interfere with Endosomal Localization and AP-3 Complex Association |
| title_sort | rnf13 dileucine motif variants l311s and l312p interfere with endosomal localization and ap 3 complex association |
| topic | RNF13 AP-3 complex endosomes lysosomes intracellular trafficking genetic mutations |
| url | https://www.mdpi.com/2073-4409/10/11/3063 |
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