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; D’Ambrosio, Chiara; Domenis, Rossana; Comelli, Marina; Delneri, Daniela; Scaloni, Andrea; Altieri, Fabio; Mavelli, Irene; Quadrifoglio, Franco; Kelley, Mark R.; Tell, Gianluca

doi:10.1091/mbc.e11-05-0391

Cited by 67 publications

(89 citation statements)

References 77 publications

(155 reference statements)

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“…Although the KD cells eventually die, exhibiting the typical growth arrest and apoptosis, this model has been a valuable tool for delineating the contribution of specific APE1 functions. Consistent with the previous work, KD cells complemented with a nuclease-deficient human APE1 cDNA harboring an H309N mutation did not survive, confirming the essential nature of the DNA repair activities of the enzyme (213). However, in contrast to what the Mitra laboratory observed (see above), the C65S redox regulatory mutant did not rescue the inviability of the KD cells; whereas the non-acetylatable K6R/K7R mutant provided near wild-type complementation for cell growth.…”

supporting

confidence: 84%

“…Moreover, a recent study found that full-length APE1 resides mainly in the mitochondrial intermembrane space (213), a localization pattern that appears distinct from other mtBER proteins, which are associated with mtDNA in the matrix (198). In this work (213), APE1 was shown to interact, in a proximity ligation assay, with the oxidoreductase Mia40, which plays a key role in oxidative protein folding within the mitochondrial inter-membrane space. Thus, the current data suggest a model in which a presently unknown PTM of full-length APE1 is necessary to reveal the ''masked'' MTS within the C-terminus and enable mitochondrial-specific protein interactions that direct mitochondrial localization.…”

Section: Intra-cellular Targeting and Mitochondrial Functionmentioning

confidence: 58%

“…As noted earlier, Vascotto et al showed that the redox mutant C65S is unable to rescue the inviability of the KD HeLa cell line (213). However, they also found that this mutant was unable to counteract the decreased mitochondrial membrane potential, that is, the mitochondrial dysfunction, of the APE1-deficient cells.…”

Section: Intra-cellular Targeting and Mitochondrial Functionmentioning

confidence: 82%

“…For example, as described in the previous section, Vascotto et al found that the redox state of C65 can control the mitochondrial localization of APE1 (213). In addition, one study showed that S-nitrosoglutathion, a nitric oxide donor and S-nitrosating agent, can induce APE1 nuclear export, with this nuclear to cytoplasmic translocation being mediated by S-nitrosation of residues C93 and C310 (165).…”

Section: Intra-cellular Traffickingmentioning

confidence: 87%

“…2), via a site-specific cleavage event that might engage a mitochondrial matrix peptidase. However, given the many studies which have reported full-length APE1 in the mitochondrial fraction [see for instance (203,213)] and the evidence that the N-terminal portion of APE1 is susceptible to degradation (182,184), it seems likely that non-specific proteolysis occurred during the fractionation steps, leading to the creation of the 33 amino-acid N-terminal truncated form. Since there is no consensus mitochondrial targeting sequence (MTS) in APE1 based on bioinformatics scrutiny, Li et al (118) screened for interactions between a series of peptides that span the length of the APE1 protein and three translocases of the outer mitochondrial membrane.…”

Section: Intra-cellular Targeting and Mitochondrial Functionmentioning

confidence: 99%

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Human Apurinic/Apyrimidinic Endonuclease 1

2014

Antioxidants & Redox Signaling

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Significance: Human apurinic/apyrimidinic endonuclease 1 (APE1, also known as REF-1) was isolated based on its ability to cleave at AP sites in DNA or activate the DNA binding activity of certain transcription factors. We review herein topics related to this multi-functional DNA repair and stress-response protein. Recent Advances: APE1 displays homology to Escherichia coli exonuclease III and is a member of the divalent metal-dependent a/b fold-containing phosphoesterase superfamily of enzymes. APE1 has acquired distinct active site and loop elements that dictate substrate selectivity, and a unique N-terminus which at minimum imparts nuclear targeting and interaction specificity. Additional activities ascribed to APE1 include 3¢-5¢ exonuclease, 3¢-repair diesterase, nucleotide incision repair, damaged or site-specific RNA cleavage, and multiple transcription regulatory roles. Critical Issues: APE1 is essential for mouse embryogenesis and contributes to cell viability in a genetic background-dependent manner. Haploinsufficient APE1 +/ -mice exhibit reduced survival, increased cancer formation, and cellular/tissue hyper-sensitivity to oxidative stress, supporting the notion that impaired APE1 function associates with disease susceptibility. Although abnormal APE1 expression/localization has been seen in cancer and neuropathologies, and impaired-function variants have been described, a causal link between an APE1 defect and human disease remains elusive. Future Directions: Ongoing efforts aim at delineating the biological role(s) of the different APE1 activities, as well as the regulatory mechanisms for its intra-cellular distribution and participation in diverse molecular pathways. The determination of whether APE1 defects contribute to human disease, particularly pathologies that involve oxidative stress, and whether APE1 small-molecule regulators have clinical utility, is central to future investigations. Antioxid. Redox Signal. 20,

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supporting

confidence: 84%

Section: Intra-cellular Targeting and Mitochondrial Functionmentioning

confidence: 58%

Section: Intra-cellular Targeting and Mitochondrial Functionmentioning

confidence: 82%

Section: Intra-cellular Traffickingmentioning

confidence: 87%

Section: Intra-cellular Targeting and Mitochondrial Functionmentioning

confidence: 99%

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Human Apurinic/Apyrimidinic Endonuclease 1

2014

Antioxidants & Redox Signaling

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224

221

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Polyubiquitination of apurinic/apyrimidinic endonuclease 1 by Parkin

Scott

Wicker

Rangaswamy

et al. 2016

Molecular Carcinogenesis

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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. While 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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DNA Repair and Mutagenesis in Vertebrate Mitochondria: Evidence for Asymmetric DNA Strand Inheritance

Matkarimov

Saparbaev

2020

Advances in Experimental Medicine and Biology

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A variety of endogenous and exogenous factors induce chemical and structural alterations in cellular DNA in addition to the errors occurring throughout DNA synthesis. These types of DNA damage are cytotoxic, miscoding or both and are believed to be at the origin of cancer and other age-related diseases. A human cell, aside from nuclear DNA, contains thousands of copies of mitochondrial DNA (mtDNA), a double-stranded, circular molecule of 16,569 bp. It has been proposed that mtDNA is a critical target of reactive oxygen species: by-products of oxidative phosphorylation that are generated in the organelle during aerobic respiration. Indeed, oxidative damage to mtDNA is more extensive and persistent as compared to that to nuclear DNA. Although transversions are the hallmark of mutations induced by reactive oxygen species, paradoxically, the majority of mtDNA mutations that occur during ageing and cancer are transitions. Furthermore, these mutations show a striking strand orientation bias: T→C/G→A transitions preferentially occur on the light strand, whereas C→T/A→G on the heavy strand of mtDNA. Here, we propose that the majority of mtDNA progenies, created after multiple rounds of DNA replication, are derived from the heavy strand only, owing to asymmetric replication of the DNA strand anchored to the inner membrane via the D-loop structure.

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

Cited by 67 publications

References 77 publications

Human Apurinic/Apyrimidinic Endonuclease 1

Human Apurinic/Apyrimidinic Endonuclease 1

Polyubiquitination of apurinic/apyrimidinic endonuclease 1 by Parkin

DNA Repair and Mutagenesis in Vertebrate Mitochondria: Evidence for Asymmetric DNA Strand Inheritance

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