Loss of KEAP1 Causes an Accumulation of Nondegradative Organelles
KEAP1 is a cytoplasmic protein that functions as an adaptor for the Cullin-3-based ubiquitin E3 ligase system, which regulates the degradation of many proteins, including NFE2L2/NRF2 and p62/SQSTM1. Loss of KEAP1 leads to an accumulation of protein ubiquitin aggregates and defective autophagy. To be...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-07-01
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| Series: | Antioxidants |
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| Online Access: | https://www.mdpi.com/2076-3921/11/7/1398 |
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| author | Elisabet Uribe-Carretero Guadalupe Martinez-Chacón Sokhna M. S. Yakhine-Diop Gema Duque-González Mario Rodríguez-Arribas Eva Alegre-Cortés Marta Paredes-Barquero Saray Canales-Cortés Elisa Pizarro-Estrella Antonio Cuadrado Rosa Ana González-Polo José M. Fuentes Mireia Niso-Santano |
| author_facet | Elisabet Uribe-Carretero Guadalupe Martinez-Chacón Sokhna M. S. Yakhine-Diop Gema Duque-González Mario Rodríguez-Arribas Eva Alegre-Cortés Marta Paredes-Barquero Saray Canales-Cortés Elisa Pizarro-Estrella Antonio Cuadrado Rosa Ana González-Polo José M. Fuentes Mireia Niso-Santano |
| author_sort | Elisabet Uribe-Carretero |
| collection | DOAJ |
| description | KEAP1 is a cytoplasmic protein that functions as an adaptor for the Cullin-3-based ubiquitin E3 ligase system, which regulates the degradation of many proteins, including NFE2L2/NRF2 and p62/SQSTM1. Loss of KEAP1 leads to an accumulation of protein ubiquitin aggregates and defective autophagy. To better understand the role of KEAP1 in the degradation machinery, we investigated whether Keap1 deficiency affects the endosome-lysosomal pathway. We used KEAP1-deficient mouse embryonic fibroblasts (MEFs) and combined Western blot analysis and fluorescence microscopy with fluorometric and pulse chase assays to analyze the levels of lysosomal-endosomal proteins, lysosomal function, and autophagy activity. We found that the loss of keap1 downregulated the protein levels and activity of the cathepsin D enzyme. Moreover, KEAP1 deficiency caused lysosomal alterations accompanied by an accumulation of autophagosomes. Our study demonstrates that KEAP1 deficiency increases nondegradative lysosomes and identifies a new role for KEAP1 in lysosomal function that may have therapeutic implications. |
| first_indexed | 2024-03-09T10:23:31Z |
| format | Article |
| id | doaj.art-c5c362062c1b42fbbd0a0832c1eaaf6f |
| institution | Directory Open Access Journal |
| issn | 2076-3921 |
| language | English |
| last_indexed | 2024-03-09T10:23:31Z |
| publishDate | 2022-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Antioxidants |
| spelling | doaj.art-c5c362062c1b42fbbd0a0832c1eaaf6f2023-12-01T21:50:11ZengMDPI AGAntioxidants2076-39212022-07-01117139810.3390/antiox11071398Loss of KEAP1 Causes an Accumulation of Nondegradative OrganellesElisabet Uribe-Carretero0Guadalupe Martinez-Chacón1Sokhna M. S. Yakhine-Diop2Gema Duque-González3Mario Rodríguez-Arribas4Eva Alegre-Cortés5Marta Paredes-Barquero6Saray Canales-Cortés7Elisa Pizarro-Estrella8Antonio Cuadrado9Rosa Ana González-Polo10José M. Fuentes11Mireia Niso-Santano12Departamento de Bioquímica y Biología Molecular y Genética, Facultad de Enfermería y Terapia Ocupacional, Universidad de Extremadura, 10003 Cáceres, SpainDepartamento de Bioquímica y Biología Molecular y Genética, Facultad de Enfermería y Terapia Ocupacional, Universidad de Extremadura, 10003 Cáceres, SpainCentro de Investigación Biomédica en Red en Enfermedades Neurodegenerativas (CIBERNED), 28029 Madrid, SpainDepartamento de Bioquímica y Biología Molecular y Genética, Facultad de Enfermería y Terapia Ocupacional, Universidad de Extremadura, 10003 Cáceres, SpainDepartamento de Bioquímica y Biología Molecular y Genética, Facultad de Enfermería y Terapia Ocupacional, Universidad de Extremadura, 10003 Cáceres, SpainDepartamento de Bioquímica y Biología Molecular y Genética, Facultad de Enfermería y Terapia Ocupacional, Universidad de Extremadura, 10003 Cáceres, SpainDepartamento de Bioquímica y Biología Molecular y Genética, Facultad de Enfermería y Terapia Ocupacional, Universidad de Extremadura, 10003 Cáceres, SpainDepartamento de Bioquímica y Biología Molecular y Genética, Facultad de Enfermería y Terapia Ocupacional, Universidad de Extremadura, 10003 Cáceres, SpainDepartamento de Bioquímica y Biología Molecular y Genética, Facultad de Enfermería y Terapia Ocupacional, Universidad de Extremadura, 10003 Cáceres, SpainCentro de Investigación Biomédica en Red en Enfermedades Neurodegenerativas (CIBERNED), 28029 Madrid, SpainDepartamento de Bioquímica y Biología Molecular y Genética, Facultad de Enfermería y Terapia Ocupacional, Universidad de Extremadura, 10003 Cáceres, SpainDepartamento de Bioquímica y Biología Molecular y Genética, Facultad de Enfermería y Terapia Ocupacional, Universidad de Extremadura, 10003 Cáceres, SpainDepartamento de Bioquímica y Biología Molecular y Genética, Facultad de Enfermería y Terapia Ocupacional, Universidad de Extremadura, 10003 Cáceres, SpainKEAP1 is a cytoplasmic protein that functions as an adaptor for the Cullin-3-based ubiquitin E3 ligase system, which regulates the degradation of many proteins, including NFE2L2/NRF2 and p62/SQSTM1. Loss of KEAP1 leads to an accumulation of protein ubiquitin aggregates and defective autophagy. To better understand the role of KEAP1 in the degradation machinery, we investigated whether Keap1 deficiency affects the endosome-lysosomal pathway. We used KEAP1-deficient mouse embryonic fibroblasts (MEFs) and combined Western blot analysis and fluorescence microscopy with fluorometric and pulse chase assays to analyze the levels of lysosomal-endosomal proteins, lysosomal function, and autophagy activity. We found that the loss of keap1 downregulated the protein levels and activity of the cathepsin D enzyme. Moreover, KEAP1 deficiency caused lysosomal alterations accompanied by an accumulation of autophagosomes. Our study demonstrates that KEAP1 deficiency increases nondegradative lysosomes and identifies a new role for KEAP1 in lysosomal function that may have therapeutic implications.https://www.mdpi.com/2076-3921/11/7/1398autophagycathepsin DendosomesKEAP1LAMP1lysosomes |
| spellingShingle | Elisabet Uribe-Carretero Guadalupe Martinez-Chacón Sokhna M. S. Yakhine-Diop Gema Duque-González Mario Rodríguez-Arribas Eva Alegre-Cortés Marta Paredes-Barquero Saray Canales-Cortés Elisa Pizarro-Estrella Antonio Cuadrado Rosa Ana González-Polo José M. Fuentes Mireia Niso-Santano Loss of KEAP1 Causes an Accumulation of Nondegradative Organelles Antioxidants autophagy cathepsin D endosomes KEAP1 LAMP1 lysosomes |
| title | Loss of KEAP1 Causes an Accumulation of Nondegradative Organelles |
| title_full | Loss of KEAP1 Causes an Accumulation of Nondegradative Organelles |
| title_fullStr | Loss of KEAP1 Causes an Accumulation of Nondegradative Organelles |
| title_full_unstemmed | Loss of KEAP1 Causes an Accumulation of Nondegradative Organelles |
| title_short | Loss of KEAP1 Causes an Accumulation of Nondegradative Organelles |
| title_sort | loss of keap1 causes an accumulation of nondegradative organelles |
| topic | autophagy cathepsin D endosomes KEAP1 LAMP1 lysosomes |
| url | https://www.mdpi.com/2076-3921/11/7/1398 |
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