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Knock-in reconstitution studies reveal an unexpected role of Cys-65 in regulating APE1/Ref-1 subcellular trafficking and function
Apurinic/apyrimidinic endonuclease 1/redox factor-1 (APE1) protects cells from oxidative stress via the base excision repair pathway and as a redox transcriptional coactivator. It is required for tumor progression/metastasis, and its up-regulation is associated with cancer resistance. Loss of APE1 e...
Autores principales: | , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The American Society for Cell Biology
2011
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3192867/ https://www.ncbi.nlm.nih.gov/pubmed/21865600 http://dx.doi.org/10.1091/mbc.E11-05-0391 |
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author | Vascotto, Carlo Bisetto, Elena Li, Mengxia Zeef, Leo A. H. D'Ambrosio, Chiara Domenis, Rossana Comelli, Marina Delneri, Daniela Scaloni, Andrea Altieri, Fabio Mavelli, Irene Quadrifoglio, Franco Kelley, Mark R. Tell, Gianluca |
author_facet | Vascotto, Carlo Bisetto, Elena Li, Mengxia Zeef, Leo A. H. D'Ambrosio, Chiara Domenis, Rossana Comelli, Marina Delneri, Daniela Scaloni, Andrea Altieri, Fabio Mavelli, Irene Quadrifoglio, Franco Kelley, Mark R. Tell, Gianluca |
author_sort | Vascotto, Carlo |
collection | PubMed |
description | Apurinic/apyrimidinic endonuclease 1/redox factor-1 (APE1) protects cells from oxidative stress via the base excision repair pathway and as a redox transcriptional coactivator. It is required for tumor progression/metastasis, and its up-regulation is associated with cancer resistance. Loss of APE1 expression causes cell growth arrest, mitochondrial impairment, apoptosis, and alterations of the intracellular redox state and cytoskeletal structure. A detailed knowledge of the molecular mechanisms regulating its different activities is required to understand the APE1 function associated with cancer development and for targeting this protein in cancer therapy. To dissect these activities, we performed reconstitution experiments by using wild-type and various APE1 mutants. Our results suggest that the redox function is responsible for cell proliferation through the involvement of Cys-65 in mediating APE1 localization within mitochondria. C65S behaves as a loss-of-function mutation by affecting the in vivo folding of the protein and by causing a reduced accumulation in the intermembrane space of mitochondria, where the import protein Mia40 specifically interacts with APE1. Treatment of cells with (E)-3-(2-[5,6-dimethoxy-3-methyl-1,4-benzoquinonyl])-2-nonyl propenoic acid, a specific inhibitor of APE1 redox function through increased Cys-65 oxidation, confirm that Cys-65 controls APE1 subcellular trafficking and provides the basis for a new role for this residue. |
format | Online Article Text |
id | pubmed-3192867 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2011 |
publisher | The American Society for Cell Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-31928672011-12-30 Knock-in reconstitution studies reveal an unexpected role of Cys-65 in regulating APE1/Ref-1 subcellular trafficking and function Vascotto, Carlo Bisetto, Elena Li, Mengxia Zeef, Leo A. H. D'Ambrosio, Chiara Domenis, Rossana Comelli, Marina Delneri, Daniela Scaloni, Andrea Altieri, Fabio Mavelli, Irene Quadrifoglio, Franco Kelley, Mark R. Tell, Gianluca Mol Biol Cell Articles Apurinic/apyrimidinic endonuclease 1/redox factor-1 (APE1) protects cells from oxidative stress via the base excision repair pathway and as a redox transcriptional coactivator. It is required for tumor progression/metastasis, and its up-regulation is associated with cancer resistance. Loss of APE1 expression causes cell growth arrest, mitochondrial impairment, apoptosis, and alterations of the intracellular redox state and cytoskeletal structure. A detailed knowledge of the molecular mechanisms regulating its different activities is required to understand the APE1 function associated with cancer development and for targeting this protein in cancer therapy. To dissect these activities, we performed reconstitution experiments by using wild-type and various APE1 mutants. Our results suggest that the redox function is responsible for cell proliferation through the involvement of Cys-65 in mediating APE1 localization within mitochondria. C65S behaves as a loss-of-function mutation by affecting the in vivo folding of the protein and by causing a reduced accumulation in the intermembrane space of mitochondria, where the import protein Mia40 specifically interacts with APE1. Treatment of cells with (E)-3-(2-[5,6-dimethoxy-3-methyl-1,4-benzoquinonyl])-2-nonyl propenoic acid, a specific inhibitor of APE1 redox function through increased Cys-65 oxidation, confirm that Cys-65 controls APE1 subcellular trafficking and provides the basis for a new role for this residue. The American Society for Cell Biology 2011-10-15 /pmc/articles/PMC3192867/ /pubmed/21865600 http://dx.doi.org/10.1091/mbc.E11-05-0391 Text en © 2011 Vascotto et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0). “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society of Cell Biology. |
spellingShingle | Articles Vascotto, Carlo Bisetto, Elena Li, Mengxia Zeef, Leo A. H. D'Ambrosio, Chiara Domenis, Rossana Comelli, Marina Delneri, Daniela Scaloni, Andrea Altieri, Fabio Mavelli, Irene Quadrifoglio, Franco Kelley, Mark R. Tell, Gianluca Knock-in reconstitution studies reveal an unexpected role of Cys-65 in regulating APE1/Ref-1 subcellular trafficking and function |
title | Knock-in reconstitution studies reveal an unexpected role of Cys-65 in regulating APE1/Ref-1 subcellular trafficking and function |
title_full | Knock-in reconstitution studies reveal an unexpected role of Cys-65 in regulating APE1/Ref-1 subcellular trafficking and function |
title_fullStr | Knock-in reconstitution studies reveal an unexpected role of Cys-65 in regulating APE1/Ref-1 subcellular trafficking and function |
title_full_unstemmed | Knock-in reconstitution studies reveal an unexpected role of Cys-65 in regulating APE1/Ref-1 subcellular trafficking and function |
title_short | Knock-in reconstitution studies reveal an unexpected role of Cys-65 in regulating APE1/Ref-1 subcellular trafficking and function |
title_sort | knock-in reconstitution studies reveal an unexpected role of cys-65 in regulating ape1/ref-1 subcellular trafficking and function |
topic | Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3192867/ https://www.ncbi.nlm.nih.gov/pubmed/21865600 http://dx.doi.org/10.1091/mbc.E11-05-0391 |
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