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Pharmacological chaperone-rescued cystic fibrosis CFTR-F508del mutant overcomes PRAF2-gated access to endoplasmic reticulum exit sites

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Abstract

The endoplasmic reticulum exit of some polytopic plasma membrane proteins (PMPs) is controlled by arginin-based retention motifs. PRAF2, a gatekeeper which recognizes these motifs, was shown to retain the GABAB-receptor GB1 subunit in the ER. We report that PRAF2 can interact on a stoichiometric basis with both wild type and mutant F508del Cystic Fibrosis (CF) Transmembrane Conductance Regulator (CFTR), preventing the access of newly synthesized cargo to ER exit sites. Because of its lower abundance, compared to wild-type CFTR, CFTR-F508del recruitment into COPII vesicles is suppressed by the ER-resident PRAF2. We also demonstrate that some pharmacological chaperones that efficiently rescue CFTR-F508del loss of function in CF patients target CFTR-F508del retention by PRAF2 operating with various mechanisms. Our findings open new therapeutic perspectives for diseases caused by the impaired cell surface trafficking of mutant PMPs, which contain RXR-based retention motifs that might be recognized by PRAF2.

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The datasets generated during and/or analyzed during the current study are available as supplemental tables and figures.

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Acknowledgements

The authors thank the IMAG’IC microscopy facility of the Institut Cochin, supported by the National Infrastructure France Bio-Imaging (grant ANR-10-INBS-04) and IBISA consortium, for its assistance in imaging experiments.

Funding

This work was supported by institutional grants from the Institut National de Santé et de la Recherché Médicale (INSERM), the Centre National de la Recherche Scientifique (CNRS) the Université Paris Cité, by the Agence Nationale pour la Recherche (ANR): ANR-18-CE14-0004-02 grant to SM and AH, the “Vaincre la Mucoviscidose” RF20180502279 grant to SM and AE and the “Association pour l’Aide à la Recherche contre la Mucoviscidose (AARM)” to IS-G.

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  1. Kusumika Saha

    Present address: Division of Cardiovascular Medicine, Brigham and Women’s Hospital, Boston, MA, 02115, USA

  2. Stéphane Doly

    Present address: Université Clermont Auvergne, INSERM U1107, NEURO-DOL, 63000, Clermont-Ferrand, France

  3. Alexandre Hinzpeter and Stefano Marullo are equally contributing senior authors.

Authors and Affiliations

  1. Université Paris Cité, Institut Cochin, INSERM U1016, CNRS UMR 8104, 75014, Paris, France

    Kusumika Saha, Stéphane Doly, Thomas Guilbert, Mark G. H. Scott, Hervé Enslen & Stefano Marullo

  2. Institut-Necker-Enfants-Malades, Université Paris Cité, INSERM U1151, CNRS UMR 8253, 75015, Paris, France

    Benoit Chevalier, Nesrine Baatallah, Iwona Pranke, Aleksander Edelman, Isabelle Sermet-Gaudelus & Alexandre Hinzpeter

  3. Proteomic Facility, Institut-Necker-Enfants-Malades, Université Paris Cité, 75015, Paris, France

    Chiara Guerrera & Cérina Chuon

  4. Centre de Référence Maladies Rares Mucoviscidose et Maladies de CFTR, Hôpital Necker Enfants Malades, European Reference National (ERN) Lung Center, Paris, France

    Isabelle Sermet-Gaudelus

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Contributions

SM, KS and AH contributed to the study conception and design. Material preparation, data collection and/or analysis were performed by KS, BC, SD, NB, TG, IP, MGHS, HE, CG, CC, AE, IS-G, AH and SM. The first draft of the manuscript was written by SM, and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

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Correspondence to Stefano Marullo.

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Saha, K., Chevalier, B., Doly, S. et al. Pharmacological chaperone-rescued cystic fibrosis CFTR-F508del mutant overcomes PRAF2-gated access to endoplasmic reticulum exit sites. Cell. Mol. Life Sci. 79, 530 (2022). https://doi.org/10.1007/s00018-022-04554-1

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