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Title: APE2 Zf-GRF facilitates 3'-5' resection of DNA damage following oxidative stress

Journal Article · · Proceedings of the National Academy of Sciences of the United States of America

The Xenopus laevis APE2 (apurinic/apyrimidinic endonuclease 2) nuclease participates in 3'-5' nucleolytic resection of oxidative DNA damage and activation of the ATR-Chk1 DNA damage response (DDR) pathway via ill-defined mechanisms. Here we report that APE2 resection activity is regulated by DNA interactions in its Zf-GRF domain, a region sharing high homology with DDR proteins Topoisomerase 3α (TOP3α) and NEIL3 (Nei-like DNA glycosylase 3), as well as transcription and RNA regulatory proteins, such as TTF2 (transcription termination factor 2), TFIIS, and RPB9. Biochemical and NMR results establish the nucleic acid-binding activity of the Zf-GRF domain. Moreover, an APE2 Zf-GRF X-ray structure and small-angle X-ray scattering analyses show that the Zf-GRF fold is typified by a crescent-shaped ssDNA binding claw that is flexibly appended to an APE2 endonuclease/exonuclease/phosphatase (EEP) catalytic core. Structure-guided Zf-GRF mutations impact APE2 DNA binding and 3'-5' exonuclease processing, and also prevent efficient APE2-dependent RPA recruitment to damaged chromatin and activation of the ATR-Chk1 DDR pathway in response to oxidative stress in Xenopus egg extracts. Collectively, our data unveil the APE2 Zf-GRF domain as a nucleic acid interaction module in the regulation of a key single-strand break resection function of APE2, and also reveal topologic similarity of the Zf-GRF to the zinc ribbon domains of TFIIS and RPB9.

Research Organization:
Argonne National Laboratory (ANL), Argonne, IL (United States)
Sponsoring Organization:
USDOE Office of Science (SC), Basic Energy Sciences (BES); USDOE Office of Science (SC), Biological and Environmental Research (BER); Univ. of North Carolina at Charlotte; National Inst. of General Medical Sciences of the National Inst. of Health (NIH)
Grant/Contract Number:
W-31-109-Eng-38; R15 GM101571; R15 GM114713; R01 GM105404; 1Z01ES102765; 1Z01ES050111
OSTI ID:
1343137
Journal Information:
Proceedings of the National Academy of Sciences of the United States of America, Vol. 114, Issue 2; ISSN 0027-8424
Publisher:
National Academy of SciencesCopyright Statement
Country of Publication:
United States
Language:
ENGLISH
Citation Metrics:
Cited by: 37 works
Citation information provided by
Web of Science

References (61)

Causes and consequences of replication stress journal December 2013
Sticky fingers: zinc-fingers as protein-recognition motifs journal February 2007
The DNA Damage Response: Making It Safe to Play with Knives journal October 2010
Expression, Functionality, and Localization of Apurinic/Apyrimidinic Endonucleases in Replicative and Non-Replicative Forms of Trypanosoma cruzi: AP Endonucleases, DNA Repair, and T. cruzi Survival journal December 2013
Interaction of apurinic/apyrimidinic endonuclease 2 (Apn2) with Myh1 DNA glycosylase in fission yeast journal March 2014
Determinants in nuclease specificity of ape1 and ape2, human homologues of Escherichia coli exonuclease III journal February 2002
Human APE2 protein is mostly localized in the nuclei and to some extent in the mitochondria, while nuclear APE2 is partly associated with proliferating cell nuclear antigen journal June 2001
Sensing DNA Damage Through ATRIP Recognition of RPA-ssDNA Complexes journal June 2003
Role of PCNA-dependent stimulation of 3'-phosphodiesterase and 3'-5' exonuclease activities of human Ape2 in repair of oxidative DNA damage journal May 2009
TopBP1 and DNA polymerase-α directly recruit the 9-1-1 complex to stalled DNA replication forks journal March 2009
Functional interplay between ATM/ATR-mediated DNA damage response and DNA repair pathways in oxidative stress journal June 2014
Protein mimicry of DNA and pathway regulation journal December 2005
C-terminal domains of Escherichia coli topoisomerase I belong to the zinc-ribbon superfamily journal June 2000
The DNA-damage response in human biology and disease journal October 2009
Actions of Aprataxin in Multiple DNA Repair Pathways journal February 2007
TopBP1 Activates the ATR-ATRIP Complex journal March 2006
The major human abasic endonuclease: formation, consequences and repair of abasic lesions in DNA journal May 2001
ATR: an essential regulator of genome integrity journal July 2008
ATM and ATR Substrate Analysis Reveals Extensive Protein Networks Responsive to DNA Damage journal May 2007
Structure of a new nucleic-acid-binding motif in eukaryotic transcriptional elongation factor TFIIS journal September 1993
A novel human DNA glycosylase that removes oxidative DNA damage and is homologous to Escherichia coli endonuclease VIII journal July 2002
The Many Functions of APE1/Ref-1: Not Only a DNA Repair Enzyme journal March 2009
Oxidative genome damage and its repair: Implications in aging and neurodegenerative diseases journal April 2012
Recognition and repair of chemically heterogeneous structures at DNA ends: Detection and Repair of Abnormal DNA Termini journal August 2014
Apurinic Endonuclease Activity of Yeast Apn2 Protein journal May 2000
Surviving the Breakup: The DNA Damage Checkpoint journal December 2006
DNA damage activates ATM through intermolecular autophosphorylation and dimer dissociation journal January 2003
Base Excision Repair journal April 2013
Checking on DNA damage in S phase journal October 2004
Zinc Fingers--Folds for Many Occasions journal December 2002
Human Ape2 protein has a 3'-5' exonuclease activity that acts preferentially on mismatched base pairs journal January 2006
Abasic sites in DNA: repair and biological consequences in Saccharomyces cerevisiae journal January 2004
Apurinic/Apyrimidinic Endonuclease 2 Is Necessary for Normal B Cell Development and Recovery of Lymphoid Progenitors after Chemotherapeutic Challenge journal January 2011
Structure of an aprataxin–DNA complex with insights into AOA1 neurodegenerative disease journal October 2011
Base Excision Repair and its Role in Maintaining Genome Stability journal January 2008
Zinc finger peptides for the regulation of gene expression journal October 1999
AP endonucleases process 5-methylcytosine excision intermediates during active DNA demethylation in Arabidopsis journal September 2014
Identification of APN2, the Saccharomyces cerevisiae homolog of the major human AP endonuclease HAP1, and its role in the repair of abasic sites journal October 1998
Redefining Oxidative Stress journal September 2006
Direct requirement for Xmus101 in ATR-mediated phosphorylation of Claspin bound Chk1 during checkpoint signaling journal April 2006
APE2 is required for ATR-Chk1 checkpoint activation in response to oxidative stress journal June 2013
ATM Activation by DNA Double-Strand Breaks Through the Mre11-Rad50-Nbs1 Complex journal April 2005
D2P2: database of disordered protein predictions journal November 2012
Protein mimicry of DNA from crystal structures of the uracil-DNA glycosylase inhibitor protein and its complex with Escherichia coli uracil-DNA glycosylase 1 1Edited by D. C. Rees journal March 1999
Second human protein with homology to theEscherichia coli abasic endonuclease exonuclease III journal January 2000
Aprataxin resolves adenylated RNA–DNA junctions to maintain genome integrity journal December 2013
Oxidative Stress, Bone Marrow Failure, and Genome Instability in Hematopoietic Stem Cells journal January 2015
Stimulation of 3'->5' Exonuclease and 3'-Phosphodiesterase Activities of Yeast Apn2 by Proliferating Cell Nuclear Antigen journal September 2002
Chk1 in the DNA damage response: conserved roles from yeasts to mammals journal August 2004
The Rad9-Hus1-Rad1 (9-1-1) clamp activates checkpoint signaling via TopBP1 journal June 2007
Growth retardation and dyslymphopoiesis accompanied by G2/M arrest in APEX2-null mice journal December 2004
Oxidative Stress and Its Significant Roles in Neurodegenerative Diseases and Cancer journal December 2014
Structural basis of RNA polymerase II backtracking, arrest and reactivation journal February 2011
Study of the DNA Damage Checkpoint using Xenopus Egg Extracts journal January 2012
Zinc finger proteins: getting a grip on RNA journal February 2005
Differential expression of APE1 and APE2 in germinal centers promotes error-prone repair and A:T mutations during somatic hypermutation journal June 2014
Structure of a new nucleic-acid-binding motif in eukaryotic transcriptional elongation factor TFIIS journal July 1995
Cancer genome datamining and functional genetic analysis implicate mechanisms of ATM/ATR dysfunction underpinning carcinogenesis journal March 2021
Base Excision Repair journal March 2004
ATM Activation by DNA Double-Strand Breaks Through the Mre 11-Rad50-Nbs1 Complex journal January 2007
Base Excision Repair journal March 2015

Cited By (12)

Identification of retinal homeobox ( rax ) gene -dependent genes by a microarray approach: The DNA endoglycosylase neil3 is a major downstream component of the rax genetic pathway: Microarray Analysis of RAX Downstream Genes journal November 2018
TRAIP is a master regulator of DNA interstrand crosslink repair journal March 2019
Resolution of a complex crisis at DNA 3′ termini journal April 2019
APE1 senses DNA single-strand breaks for repair and signaling journal December 2019
CiAPEX2 and CiP0, candidates of AP endonucleases in Ciona intestinalis, have 3′-5′ exonuclease activity and contribute to protection against oxidative stress journal December 2017
Multiple Functions of Long Non‐Coding RNAs in Oxidative Stress, DNA Damage Response and Cancer Progression journal August 2017
Structure and regulation of ZCCHC4 in m6A-methylation of 28S rRNA journal November 2019
Apn2 resolves blocked 3′ ends and suppresses Top1-induced mutagenesis at genomic rNMP sites journal February 2019
Distinct roles of XRCC1 in genome integrity in Xenopus egg extracts journal December 2019
APE2 promotes DNA damage response pathway from a single-strand break journal January 2018
Single-Strand Break End Resection in Genome Integrity: Mechanism and Regulation by APE2 journal August 2018
Molecular snapshots of APE1 proofreading mismatches and removing DNA damage journal January 2018

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