Keratin 8 is a scaffolding and regulatory protein of ERAD complexes
| Autor: | Iwona Maria Pranke, Benoit Chevalier, Aiswarya Premchandar, Nesrine Baatallah, Kamil F. Tomaszewski, Sara Bitam, Danielle Tondelier, Anita Golec, Jan Stolk, Gergely L. Lukacs, Pieter S. Hiemstra, Michal Dadlez, David A. Lomas, James A. Irving, Agnes Delaunay-Moisan, Eelco van Anken, Alexandre Hinzpeter, Isabelle Sermet-Gaudelus, Aleksander Edelman |
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| Jazyk: | angličtina |
| Rok vydání: | 2022 |
| Předmět: |
Pharmacology
Synthetic lethality Keratin-8 Ubiquitin-Protein Ligases Cystic Fibrosis Transmembrane Conductance Regulator Endoplasmic Reticulum-Associated Degradation Cell Biology Epithelium Cellular and Molecular Neuroscience Protein-protein interaction Intermediary filaments Humans Molecular Medicine Protein complexes fractionation Molecular Biology Cytoskeleton HeLa Cells Transcription Factors |
| Zdroj: | Cellular and Molecular Life Sciences, 79(9). SPRINGER BASEL AG |
| Popis: | Early recognition and enhanced degradation of misfolded proteins by the endoplasmic reticulum (ER) quality control and ER-associated degradation (ERAD) cause defective protein secretion and membrane targeting, as exemplified for Z-alpha 1 antitrypsin (Z-A1AT), responsible for alpha-1-antitrypsin deficiency (A1ATD) and F508del-CFTR (cystic fibrosis transmembrane conductance regulator) responsible for cystic fibrosis (CF).Prompted by our previous observation that decreasing Keratin 8 (K8) expression increased trafficking of F508del-CFTR to the plasma membrane, we investigated whether K8 impacts trafficking of soluble misfolded Z-A1AT protein. The subsequent goal of this study was to elucidate the mechanism underlying the K8-dependent regulation of protein trafficking, focusing on the ERAD pathway.The results show that diminishing K8 concentration in HeLa cells enhances secretion of both Z-A1AT and wild type (WT) A1AT with a 13-fold and 4-fold increase, respectively. K8 down-regulation triggers ER failure and cellular apoptosis when ER stress is jointly elicited by conditional expression of the μs heavy chains, as previously shown for Hrd1 knock-out. Simultaneous K8 silencing and Hrd1 knock-out did not show any synergistic effect, consistent with K8 acting in the Hrd1-governed ERAD step. Fractionation experiments reveal that K8 is recruited to ERAD complexes containing Derlin2, Sel1 and Hrd1 proteins upon expression of Z/WT-A1AT and F508del-CFTR. Treatment of the cells with c407, a small molecule inhibiting K8 interaction, decreases K8 and Derlin2 recruitment to high-order ERAD complexes. This was associated with increased Z-A1AT secretion in both HeLa and Z-homozygous A1ATD patients’ respiratory cells. Overall, we provide evidence that K8 acts as an ERAD modulator. It may play a scaffolding protein role for early-stage ERAD complexes, regulating Hrd1-governed retrotranslocation initiation/ubiquitination processes. Targeting K8-containing ERAD complexes is an attractive strategy for the pharmacotherapy of A1ATD. |
| Databáze: | OpenAIRE |
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