Apurinic/apyrimidinic endonuclease 1 (APE1) is an essential protein crucial for repair of oxidized DNA damage not only in genomic DNA but also in mitochondrial DNA. Parkin, a tumor suppressor and Parkinson's disease (PD) associated gene, is an E3 ubiquitin ligase crucial for mitophagy. Although DNA damage is known to induce mitochondrial stress, Parkin's role in regulating DNA repair proteins has not been elucidated. In this study, we examined the possibility of Parkin‐dependent ubiquitination of APE1. Ectopically expressed APE1 was degraded by Parkin and PINK1 via polyubiquitination in mouse embryonic fibroblast cells. PD‐causing mutations in Parkin and PINK1 abrogated APE1 ubiquitination. Interaction of APE1 with Parkin was observed by co‐immunoprecipitation, proximity ligation assay, and co‐localization in the cytoplasm. N‐terminal deletion of 41 amino acid residues in APE1 significantly reduced the Parkin‐dependent APE1 degradation. These results suggested that Parkin directly ubiquitinated N‐terminal Lys residues in APE1 in the cytoplasm. Modulation of Parkin and PINK1 activities under mitochondrial or oxidative stress caused moderate but statistically significant decrease of endogenous APE1 in human cell lines including SH‐SY5Y, HEK293, and A549 cells. Analyses of glioblastoma tissues showed an inverse relation between the expression levels of APE1 and Parkin. These results suggest that degradation of endogenous APE1 by Parkin occur when cells are stressed to activate Parkin, and imply a role of Parkin in maintaining the quality of APE1, and loss of Parkin may contribute to elevated APE1 levels in glioblastoma.
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This study was supported by NCI grant CA098664 (T.I.) and Cancer Center Supporting Grant (P30CA17758) by the Markey Cancer Center at University of Kentucky.
Additional Supporting Information may be found in the online version of this article.
Scott, Timothy L.; Wicker, Christina A.; Suganya, Rangaswamy; Dhar, Bithika; Pittman, Thomas A.; Horbinski, Craig; and Izumi, Tadahide, "Polyubiquitination of Apurinic/Apyrimidinic Endonuclease 1 by Parkin" (2017). Toxicology and Cancer Biology Faculty Publications. 90.