Endosomal chloride/proton exchangers need inhibitory TMEM9 β-subunits for regulation and prevention of disease-causing overactivity

Abstract The function of endosomes critically depends on their ion homeostasis. A crucial role of luminal Cl−, in addition to that of H+, is increasingly recognized. Both ions are transported by five distinct endolysosomal CLC chloride/proton exchangers. Dysfunction of each of these transporters ent...

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Main Authors: Rosa Planells-Cases, Viktoriia Vorobeva, Sumanta Kar, Franziska W. Schmitt, Uwe Schulte, Marina Schrecker, Richard K. Hite, Bernd Fakler, Thomas J. Jentsch
Format: Article
Language:English
Published: Nature Portfolio 2025-04-01
Series:Nature Communications
Online Access:https://doi.org/10.1038/s41467-025-58546-3
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author Rosa Planells-Cases
Viktoriia Vorobeva
Sumanta Kar
Franziska W. Schmitt
Uwe Schulte
Marina Schrecker
Richard K. Hite
Bernd Fakler
Thomas J. Jentsch
author_facet Rosa Planells-Cases
Viktoriia Vorobeva
Sumanta Kar
Franziska W. Schmitt
Uwe Schulte
Marina Schrecker
Richard K. Hite
Bernd Fakler
Thomas J. Jentsch
author_sort Rosa Planells-Cases
collection DOAJ
description Abstract The function of endosomes critically depends on their ion homeostasis. A crucial role of luminal Cl−, in addition to that of H+, is increasingly recognized. Both ions are transported by five distinct endolysosomal CLC chloride/proton exchangers. Dysfunction of each of these transporters entails severe disease. Here we identified TMEM9 and TMEM9B as obligatory β-subunits for endosomal ClC-3, ClC-4, and ClC-5. Mice lacking both β-subunits displayed severely reduced levels of all three CLCs and died embryonically or shortly after birth. TMEM9 proteins regulate trafficking of their partners. Surprisingly, they also strongly inhibit CLC ion transport. Tonic inhibition enables the regulation of CLCs and prevents toxic Cl− accumulation and swelling of endosomes. Inhibition requires a carboxy-terminal TMEM9 domain that interacts with CLCs at multiple sites. Disease-causing CLCN mutations that weaken inhibition by TMEM9 proteins cause a pathogenic gain of ion transport. Our work reveals the need to suppress, in a regulated manner, endolysosomal chloride/proton exchange. Several aspects of endosomal ion transport must be revised.
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spelling doaj-art-c2b325bfc39f4a33ae9caa89bf02789b2025-08-20T03:07:43ZengNature PortfolioNature Communications2041-17232025-04-0116111710.1038/s41467-025-58546-3Endosomal chloride/proton exchangers need inhibitory TMEM9 β-subunits for regulation and prevention of disease-causing overactivityRosa Planells-Cases0Viktoriia Vorobeva1Sumanta Kar2Franziska W. Schmitt3Uwe Schulte4Marina Schrecker5Richard K. Hite6Bernd Fakler7Thomas J. Jentsch8Leibniz-Forschungsinstitut für Molekulare Pharmakologie (FMP)Leibniz-Forschungsinstitut für Molekulare Pharmakologie (FMP)Leibniz-Forschungsinstitut für Molekulare Pharmakologie (FMP)Leibniz-Forschungsinstitut für Molekulare Pharmakologie (FMP)Institute of Physiology, Faculty of Medicine, University of FreiburgStructural Biology Program, Memorial Sloan Kettering Cancer CenterStructural Biology Program, Memorial Sloan Kettering Cancer CenterInstitute of Physiology, Faculty of Medicine, University of FreiburgLeibniz-Forschungsinstitut für Molekulare Pharmakologie (FMP)Abstract The function of endosomes critically depends on their ion homeostasis. A crucial role of luminal Cl−, in addition to that of H+, is increasingly recognized. Both ions are transported by five distinct endolysosomal CLC chloride/proton exchangers. Dysfunction of each of these transporters entails severe disease. Here we identified TMEM9 and TMEM9B as obligatory β-subunits for endosomal ClC-3, ClC-4, and ClC-5. Mice lacking both β-subunits displayed severely reduced levels of all three CLCs and died embryonically or shortly after birth. TMEM9 proteins regulate trafficking of their partners. Surprisingly, they also strongly inhibit CLC ion transport. Tonic inhibition enables the regulation of CLCs and prevents toxic Cl− accumulation and swelling of endosomes. Inhibition requires a carboxy-terminal TMEM9 domain that interacts with CLCs at multiple sites. Disease-causing CLCN mutations that weaken inhibition by TMEM9 proteins cause a pathogenic gain of ion transport. Our work reveals the need to suppress, in a regulated manner, endolysosomal chloride/proton exchange. Several aspects of endosomal ion transport must be revised.https://doi.org/10.1038/s41467-025-58546-3
spellingShingle Rosa Planells-Cases
Viktoriia Vorobeva
Sumanta Kar
Franziska W. Schmitt
Uwe Schulte
Marina Schrecker
Richard K. Hite
Bernd Fakler
Thomas J. Jentsch
Endosomal chloride/proton exchangers need inhibitory TMEM9 β-subunits for regulation and prevention of disease-causing overactivity
Nature Communications
title Endosomal chloride/proton exchangers need inhibitory TMEM9 β-subunits for regulation and prevention of disease-causing overactivity
title_full Endosomal chloride/proton exchangers need inhibitory TMEM9 β-subunits for regulation and prevention of disease-causing overactivity
title_fullStr Endosomal chloride/proton exchangers need inhibitory TMEM9 β-subunits for regulation and prevention of disease-causing overactivity
title_full_unstemmed Endosomal chloride/proton exchangers need inhibitory TMEM9 β-subunits for regulation and prevention of disease-causing overactivity
title_short Endosomal chloride/proton exchangers need inhibitory TMEM9 β-subunits for regulation and prevention of disease-causing overactivity
title_sort endosomal chloride proton exchangers need inhibitory tmem9 β subunits for regulation and prevention of disease causing overactivity
url https://doi.org/10.1038/s41467-025-58546-3
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