{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,6]],"date-time":"2026-07-06T17:25:31Z","timestamp":1783358731345,"version":"3.54.6"},"reference-count":48,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2023,6,6]],"date-time":"2023-06-06T00:00:00Z","timestamp":1686009600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001871","name":"Portuguese Funda\u00e7\u00e3o para a Ci\u00eancia e a Tecnologia","doi-asserted-by":"publisher","award":["PTDC\/BIA-CEL\/28408\/2017"],"award-info":[{"award-number":["PTDC\/BIA-CEL\/28408\/2017"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001871","name":"Portuguese Funda\u00e7\u00e3o para a Ci\u00eancia e a Tecnologia","doi-asserted-by":"publisher","award":["UID\/MULTI\/04046\/2019"],"award-info":[{"award-number":["UID\/MULTI\/04046\/2019"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biomolecules"],"abstract":"<jats:p>Recent developments in CFTR modulator drugs have had a significant transformational effect on the treatment of individuals with Cystic Fibrosis (CF) who carry the most frequent F508del-CFTR mutation in at least one allele. However, the clinical effects of these revolutionary drugs remain limited by their inability to fully restore the plasma membrane (PM) stability of the rescued mutant channels. Here, we shed new light on the molecular mechanisms behind the reduced half-life of rescued F508del-CFTR at the PM of airway cells. We describe that YES1 protein kinase is enriched in F508del-CFTR protein PM complexes, and that its interaction with rescued channels is mediated and dependent on the adaptor protein YAP1. Moreover, we show that interference with this complex, either by depletion of one of these components or inhibiting YES1 activity, is sufficient to significantly improve the abundance and stability of modulator-rescued F508del-CFTR at the surface of airway cells. In addition, we found that this effect was mediated by a decreased phosphorylation of the scaffold protein SHC1, a key regulator of MAPK pathway activity. In fact, we showed that depletion of SHC1 or inhibition of MAPK pathway signaling was sufficient to improve rescued F508del-CFTR surface levels, whereas an ectopic increase in pathway activation downstream of SHC1, through the use of a constitutively active H-RAS protein, abrogated the stabilizing effect of YES1 inhibition on rescued F508del-CFTR. Taken together, our findings not only provide new mechanistic insights into the regulation of modulator-rescued F508del-CFTR membrane stability, but also open exciting new avenues to be further explored in CF research and treatment.<\/jats:p>","DOI":"10.3390\/biom13060949","type":"journal-article","created":{"date-parts":[[2023,6,7]],"date-time":"2023-06-07T02:28:45Z","timestamp":1686104925000},"page":"949","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["YES1 Kinase Mediates the Membrane Removal of Rescued F508del-CFTR in Airway Cells by Promoting MAPK Pathway Activation via SHC1"],"prefix":"10.3390","volume":"13","author":[{"given":"Patr\u00edcia","family":"Barros","sequence":"first","affiliation":[{"name":"Departamento de Gen\u00e9tica Humana, Instituto Nacional de Sa\u00fade Doutor Ricardo Jorge, 1649-016 Lisboa, Portugal"},{"name":"BioISI\u2014Biosystems & Integrative Sciences Institute, Faculty of Sciences, University of Lisboa, 1749-016 Lisboa, Portugal"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0160-256X","authenticated-orcid":false,"given":"Ana M.","family":"Matos","sequence":"additional","affiliation":[{"name":"Departamento de Gen\u00e9tica Humana, Instituto Nacional de Sa\u00fade Doutor Ricardo Jorge, 1649-016 Lisboa, Portugal"},{"name":"BioISI\u2014Biosystems & Integrative Sciences Institute, Faculty of Sciences, University of Lisboa, 1749-016 Lisboa, Portugal"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9379-9696","authenticated-orcid":false,"given":"Paulo","family":"Matos","sequence":"additional","affiliation":[{"name":"Departamento de Gen\u00e9tica Humana, Instituto Nacional de Sa\u00fade Doutor Ricardo Jorge, 1649-016 Lisboa, Portugal"},{"name":"BioISI\u2014Biosystems & Integrative Sciences Institute, Faculty of Sciences, University of Lisboa, 1749-016 Lisboa, Portugal"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1425-9211","authenticated-orcid":false,"given":"Peter","family":"Jordan","sequence":"additional","affiliation":[{"name":"Departamento de Gen\u00e9tica Humana, Instituto Nacional de Sa\u00fade Doutor Ricardo Jorge, 1649-016 Lisboa, Portugal"},{"name":"BioISI\u2014Biosystems & Integrative Sciences Institute, Faculty of Sciences, University of Lisboa, 1749-016 Lisboa, Portugal"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,6,6]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"701","DOI":"10.1146\/annurev.biochem.75.103004.142532","article-title":"CFTR Function and Prospects for Therapy","volume":"77","author":"Riordan","year":"2008","journal-title":"Annu. Rev. Biochem."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"23","DOI":"10.1891\/NN-2022-0007","article-title":"Cystic Fibrosis: Back to the Basics","volume":"42","author":"Myer","year":"2023","journal-title":"Neonatal Netw."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"5212","DOI":"10.1021\/acs.jmedchem.1c01897","article-title":"Proteostasis Regulators in Cystic Fibrosis: Current Development and Future Perspectives","volume":"65","author":"Brusa","year":"2022","journal-title":"J. Med. Chem."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"35825","DOI":"10.1074\/jbc.M110.131623","article-title":"The Cystic Fibrosis-Causing Mutation DeltaF508 Affects Multiple Steps in Cystic Fibrosis Transmembrane Conductance Regulator Biogenesis","volume":"285","author":"Thibodeau","year":"2010","journal-title":"J. Biol. Chem."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"426","DOI":"10.1038\/nrm1949","article-title":"New Insights into Cystic Fibrosis: Molecular Switches That Regulate CFTR","volume":"7","author":"Guggino","year":"2006","journal-title":"Nat. Rev. Mol. Cell. Biol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"2519","DOI":"10.1016\/S0140-6736(16)00576-6","article-title":"Cystic Fibrosis","volume":"388","author":"Elborn","year":"2016","journal-title":"Lancet"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"897","DOI":"10.1080\/17460441.2021.1912732","article-title":"Discovery of CFTR Modulators for the Treatment of Cystic Fibrosis","volume":"16","author":"Pedemonte","year":"2021","journal-title":"Expert. Opin. Drug. Discov."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1940","DOI":"10.1016\/S0140-6736(19)32597-8","article-title":"Efficacy and Safety of the Elexacaftor plus Tezacaftor plus Ivacaftor Combination Regimen in People with Cystic Fibrosis Homozygous for the F508del Mutation: A Double-Blind, Randomised, Phase 3 Trial","volume":"394","author":"Heijerman","year":"2019","journal-title":"Lancet"},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Purkayastha, D., Agtarap, K., Wong, K., Pereira, O., Co, J., Pakhale, S., and Kanji, S. (J. Cyst. Fibros. Off. J. Eur. Cyst. Fibros. Soc., 2023). Drug-Drug Interactions with CFTR Modulator Therapy in Cystic Fibrosis: Focus on Trikafta\u00ae\/Kaftrio\u00ae, J. Cyst. Fibros. Off. J. Eur. Cyst. Fibros. Soc., in press.","DOI":"10.1016\/j.jcf.2023.01.005"},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Capurro, V., Tomati, V., Sondo, E., Renda, M., Borrelli, A., Pastorino, C., Guidone, D., Venturini, A., Giraudo, A., and Mandrup Bertozzi, S. (2021). Partial Rescue of F508del-CFTR Stability and Trafficking Defects by Double Corrector Treatment. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22105262"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"e139983","DOI":"10.1172\/jci.insight.139983","article-title":"Allosteric Folding Correction of F508del and Rare CFTR Mutants by Elexacaftor-Tezacaftor-Ivacaftor (Trikafta) Combination","volume":"5","author":"Veit","year":"2020","journal-title":"JCI Insight"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"805","DOI":"10.1126\/science.1191542","article-title":"Peripheral Protein Quality Control Removes Unfolded CFTR from the Plasma Membrane","volume":"329","author":"Okiyoneda","year":"2010","journal-title":"Science"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1100","DOI":"10.3389\/fphar.2018.01100","article-title":"Peripheral Protein Quality Control as a Novel Drug Target for CFTR Stabilizer","volume":"9","author":"Fukuda","year":"2018","journal-title":"Front. Pharmacol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"ra48","DOI":"10.1126\/scisignal.aaa1580","article-title":"A Molecular Switch in the Scaffold NHERF1 Enables Misfolded CFTR to Evade the Peripheral Quality Control Checkpoint","volume":"8","author":"Loureiro","year":"2015","journal-title":"Sci. Signal."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"36762","DOI":"10.1074\/jbc.M508944200","article-title":"The Short Apical Membrane Half-Life of Rescued {Delta}F508-Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) Results from Accelerated Endocytosis of {Delta}F508-CFTR in Polarized Human Airway Epithelial Cells","volume":"280","author":"Brown","year":"2005","journal-title":"J. Biol. Chem."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"923","DOI":"10.1083\/jcb.200312018","article-title":"Misfolding Diverts CFTR from Recycling to Degradation: Quality Control at Early Endosomes","volume":"164","author":"Sharma","year":"2004","journal-title":"J. Cell. Biol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"555","DOI":"10.1042\/BJ20071420","article-title":"Enhanced Cell-Surface Stability of Rescued DeltaF508 Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) by Pharmacological Chaperones","volume":"410","author":"Varga","year":"2008","journal-title":"Biochem. J."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"73","DOI":"10.1091\/mbc.e09-03-0185","article-title":"Na+\/H+ Exchanger Regulatory Factor 1 Overexpression-Dependent Increase of Cytoskeleton Organization Is Fundamental in the Rescue of F508del Cystic Fibrosis Transmembrane Conductance Regulator in Human Airway CFBE41o- Cells","volume":"21","author":"Favia","year":"2010","journal-title":"Mol. Biol. Cell."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"4169","DOI":"10.1021\/bi401263h","article-title":"Stabilizing Rescued Surface-Localized \u0394F508 CFTR by Potentiation of Its Interaction with Na(+)\/H(+) Exchanger Regulatory Factor 1","volume":"53","author":"Arora","year":"2014","journal-title":"Biochemistry"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"4937","DOI":"10.1091\/mbc.e06-08-0670","article-title":"Tracking of Quantum Dot-Labeled CFTR Shows near Immobilization by C-Terminal PDZ Interactions","volume":"17","author":"Haggie","year":"2006","journal-title":"Mol. Biol. Cell."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"37634","DOI":"10.1074\/jbc.M502305200","article-title":"Ezrin Controls the Macromolecular Complexes Formed between an Adapter Protein Na+\/H+ Exchanger Regulatory Factor and the Cystic Fibrosis Transmembrane Conductance Regulator","volume":"280","author":"Li","year":"2005","journal-title":"J. Biol. Chem."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"2527","DOI":"10.1128\/MCB.01372-06","article-title":"NHERF1\/EBP50 Head-to-Tail Intramolecular Interaction Masks Association with PDZ Domain Ligands","volume":"27","author":"Morales","year":"2007","journal-title":"Mol. Cell. Biol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1527","DOI":"10.1038\/labinvest.2012.123","article-title":"NHERF1 and CFTR Restore Tight Junction Organisation and Function in Cystic Fibrosis Airway Epithelial Cells: Role of Ezrin and the RhoA\/ROCK Pathway","volume":"92","author":"Castellani","year":"2012","journal-title":"Lab. Investig. J. Tech. Methods Pathol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"13396","DOI":"10.1074\/jbc.RA119.008738","article-title":"Inhibition of Calpain 1 Restores Plasma Membrane Stability to Pharmacologically Rescued Phe508del-CFTR Variant","volume":"294","author":"Matos","year":"2019","journal-title":"J. Biol. Chem."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1371","DOI":"10.1158\/1535-7163.MCT-21-0958","article-title":"YES1: A Novel Therapeutic Target and Biomarker in Cancer","volume":"21","author":"Garmendia","year":"2022","journal-title":"Mol. Cancer Ther."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"249","DOI":"10.1007\/978-1-61779-120-8_15","article-title":"Functional Genomics Assays to Study CFTR Traffic and ENaC Function","volume":"742","author":"Appel","year":"2011","journal-title":"Methods Mol. Biol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"220","DOI":"10.1016\/S0014-5793(01)02561-3","article-title":"Green Fluorescent Protein-Based Halide Indicators with Improved Chloride and Iodide Affinities","volume":"499","author":"Galietta","year":"2001","journal-title":"FEBS Lett."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"9038","DOI":"10.1038\/srep09038","article-title":"Protein Traffic Disorders: An Effective High-Throughput Fluorescence Microscopy Pipeline for Drug Discovery","volume":"5","author":"Botelho","year":"2015","journal-title":"Sci. Rep."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"899","DOI":"10.1053\/j.gastro.2008.05.052","article-title":"B-Raf(V600E) Cooperates with Alternative Spliced Rac1b to Sustain Colorectal Cancer Cell Survival","volume":"135","author":"Matos","year":"2008","journal-title":"Gastroenterology"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"619","DOI":"10.3389\/fphar.2019.00619","article-title":"Network Biology Identifies Novel Regulators of CFTR Trafficking and Membrane Stability","volume":"10","author":"Loureiro","year":"2019","journal-title":"Front. Pharmacol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"13026","DOI":"10.1038\/s41598-018-31514-2","article-title":"Prolonged Co-Treatment with HGF Sustains Epithelial Integrity and Improves Pharmacological Rescue of Phe508del-CFTR","volume":"8","author":"Matos","year":"2018","journal-title":"Sci. Rep."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"837","DOI":"10.1016\/j.ceb.2009.09.010","article-title":"Herding Hippos: Regulating Growth in Flies and Man","volume":"21","author":"Badouel","year":"2009","journal-title":"Curr. Opin. Cell. Biol."},{"key":"ref_33","first-page":"2145","article-title":"Yes-Associated Protein (YAP65) Is a Proline-Rich Phosphoprotein That Binds to the SH3 Domain of the Yes Proto-Oncogene Product","volume":"9","author":"Sudol","year":"1994","journal-title":"Oncogene"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"879","DOI":"10.1083\/jcb.147.4.879","article-title":"Yes-Associated Protein 65 Localizes P62(c-Yes) to the Apical Compartment of Airway Epithelia by Association with EBP50","volume":"147","author":"Mohler","year":"1999","journal-title":"J. Cell. Biol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1224","DOI":"10.1016\/j.bbagen.2015.02.004","article-title":"Cigarette Smoke Exposure Reveals a Novel Role for the MEK\/ERK1\/2 MAPK Pathway in Regulation of CFTR","volume":"1850","author":"Xu","year":"2015","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Osrodek, M., and Wozniak, M. (2021). Targeting Genome Stability in Melanoma\u2014A New Approach to an Old Field. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22073485"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"292","DOI":"10.1016\/j.yexcr.2004.12.029","article-title":"Expression of Rac1b Stimulates NF-KappaB-Mediated Cell Survival and G1\/S Progression","volume":"305","author":"Matos","year":"2005","journal-title":"Exp. Cell. Res."},{"key":"ref_38","first-page":"1972","article-title":"YES Oncogenic Activity Is Specified by Its SH4 Domain and Regulates RAS\/MAPK Signaling in Colon Carcinoma Cells","volume":"5","author":"Dubois","year":"2015","journal-title":"Am. J. Cancer Res."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1435","DOI":"10.1016\/S0960-9822(96)00748-8","article-title":"The Shc Adaptor Protein Is Highly Phosphorylated at Conserved, Twin Tyrosine Residues (Y239\/240) That Mediate Protein-Protein Interactions","volume":"6","author":"Wiley","year":"1996","journal-title":"Curr. Biol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"155625","DOI":"10.1016\/j.cyto.2021.155625","article-title":"Epidermal Growth Factor Receptor and Integrins Meet Redox Signaling through P66shc and Rac1","volume":"146","author":"Mushtaq","year":"2021","journal-title":"Cytokine"},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Farinha, C.M., and Gentzsch, M. (2021). Revisiting CFTR Interactions: Old Partners and New Players. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms222413196"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"4076","DOI":"10.1128\/MCB.05152-11","article-title":"Antagonistic Regulation of Cystic Fibrosis Transmembrane Conductance Regulator Cell Surface Expression by Protein Kinases WNK4 and Spleen Tyrosine Kinase","volume":"31","author":"Mendes","year":"2011","journal-title":"Mol. Cell. Biol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"4997","DOI":"10.1007\/s00018-020-03448-4","article-title":"A SYK\/SHC1 Pathway Regulates the Amount of CFTR in the Plasma Membrane","volume":"77","author":"Loureiro","year":"2020","journal-title":"Cell. Mol. Life Sci."},{"key":"ref_44","first-page":"e22270","article-title":"Targeting the EGFR-ERK Axis Using the Compatible Solute Ectoine to Stabilize CFTR Mutant F508del","volume":"36","author":"Wellmerling","year":"2022","journal-title":"FASEB J. Off. Publ. Fed. Am. Soc. Exp. Biol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1042\/BCJ20220234","article-title":"Dynamic Regulation of RAS and RAS Signaling","volume":"480","author":"Kolch","year":"2023","journal-title":"Biochem. J."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"166","DOI":"10.1038\/nature12308","article-title":"Temporal Regulation of EGF Signalling Networks by the Scaffold Protein Shc1","volume":"499","author":"Zheng","year":"2013","journal-title":"Nature"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"13561","DOI":"10.1021\/acs.jmedchem.2c01244","article-title":"Targeting Extracellular Signal-Regulated Protein Kinase 1\/2 (ERK1\/2) in Cancer: An Update on Pharmacological Small-Molecule Inhibitors","volume":"65","author":"Fu","year":"2022","journal-title":"J. Med. Chem."},{"key":"ref_48","doi-asserted-by":"crossref","unstructured":"Vigan\u00f2, M., La Milia, M., Grassini, M.V., Pugliese, N., De Giorgio, M., and Fagiuoli, S. (2023). Hepatotoxicity of Small Molecule Protein Kinase Inhibitors for Cancer. Cancers, 15.","DOI":"10.3390\/cancers15061766"}],"container-title":["Biomolecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2218-273X\/13\/6\/949\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T19:49:14Z","timestamp":1760125754000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2218-273X\/13\/6\/949"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,6,6]]},"references-count":48,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2023,6]]}},"alternative-id":["biom13060949"],"URL":"https:\/\/doi.org\/10.3390\/biom13060949","relation":{},"ISSN":["2218-273X"],"issn-type":[{"value":"2218-273X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,6,6]]}}}