Knock-in reconstitution studies reveal an unexpected role of Cys-65 in regulating APE1/Ref-1 subcellular trafficking and function

Carlo Vascotto, Elena Bisetto, Mengxia Li, Leo A.H. Zeef, Chiara D'Ambrosio, Rossana Domenis, Marina Comelli, Daniela Delneri, Andrea Scaloni, Fabio Altieri, Irene Mavelli, Franco Quadrifoglio, Mark R. Kelley, Gianluca Tell

Research output: Contribution to journalArticle

44 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)3887-3901
Number of pages15
JournalMolecular Biology of the Cell
Volume22
Issue number20
DOIs
StatePublished - Oct 15 2011

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ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Cite this

Vascotto, C., Bisetto, E., Li, M., Zeef, L. A. H., D'Ambrosio, C., Domenis, R., Comelli, M., Delneri, D., Scaloni, A., Altieri, F., Mavelli, I., Quadrifoglio, F., Kelley, M. R., & Tell, G. (2011). Knock-in reconstitution studies reveal an unexpected role of Cys-65 in regulating APE1/Ref-1 subcellular trafficking and function. Molecular Biology of the Cell, 22(20), 3887-3901. https://doi.org/10.1091/mbc.E11-05-0391