Enigmatic role of WRN-RECQL helicase in DNA repair and its implications in cancer

Abstract

Werner (WRN) helicase belongs to the RECQL class of DNA helicases. Mutation in Werner (WRN) RECQL helicase leads to premature aging syndrome, Werner syndrome (WS), and predisposition to multiple cancers. WS patients exhibit heightened incidence of neoplasia, e.g., soft tissue sarcoma, osteosarcoma, malignant melanoma, meningioma, thyroid cancer, breast cancer, and leukemias. Extensive research on WRN helicase has revealed its important and diverse roles in DNA repair pathways, especially in double-strand break repair. Consequently, WRN deficiency is causally associated with genomic instability and cancer predispositions. In this review, we summarize recent studies unraveling the fundamental roles WRN helicase plays in DNA repair and genome stability and its implications in cancer therapy and resistance.

Keywords

RECQL / RECQ / WRN / genome instability / NHEJ / homologous recombination / aging / cancer

References

[1] Luo J.WRN protein and Werner syndrome.N Am J Med Sci (Boston)2010;3:205-7 PMCID:PMC3237395

[2] Croteau DL,Opresko PL.Human RecQ helicases in DNA repair, recombination, and replication.Annu Rev Biochem2014;83:519-52 PMCID:PMC4586249

[3] Oshima J,Monnat RJ Jr.Werner syndrome: clinical features, pathogenesis and potential therapeutic interventions.Ageing Res Rev2017;33:105-14 PMCID:PMC5025328

[4] Opresko PL,von Kobbe C,Bohr VA.Werner syndrome and the function of the Werner protein; what they can teach us about the molecular aging process.Carcinogenesis2003;24:791-802

[5] Kitano K,Hakoshima T.Crystal structure of the HRDC domain of human Werner syndrome protein, WRN.J Biol Chem2007;282:2717-28

[6] Lan L,Komatsu K.Accumulation of Werner protein at DNA double-strand breaks in human cells.J Cell Sci2005;118:4153-62

[7] Perry JJ,Holden LG.WRN exonuclease structure and molecular mechanism imply an editing role in DNA end processing.Nat Struct Mol Biol2006;13:414-22

[8] Sidorova JM,Folch A.The RecQ helicase WRN is required for normal replication fork progression after DNA damage or replication fork arrest.Cell Cycle2008;7:796-807 PMCID:PMC4362724

[9] Bugreev DV,Mazin AV.Cooperation of RAD51 and RAD54 in regression of a model replication fork.Nucleic Acids Res2011;39:2153-64 PMCID:PMC3064783

[10] Constantinou A,Karow J K.Werner’s syndrome protein (WRN) migrates Holliday junctions and co-localizes with RPA upon replication arrest.EMBO Rep2000;1:80-4 PMCID:PMC1083680

[11] Li K,Wang R.Regulation of WRN protein cellular localization and enzymatic activities by SIRT1-mediated deacetylation.J Biol Chem2008;283:7590-8

[12] Kahyo T,Goto M.Sirtuin-mediated deacetylation pathway stabilizes Werner syndrome protein.FEBS Lett2008;582:2479-83

[13] Stracker TH.The MRE11 complex: starting from the ends.Nat Rev Mol Cell Biol2011;12:90-103 PMCID:PMC3905242

[14] Hashimoto Y,Lopes M.Rad51 protects nascent DNA from Mre11-dependent degradation and promotes continuous DNA synthesis.Nat Struct Mol Biol2010;17:1305-11 PMCID:PMC4306207

[15] Lemaçon D,Quinet A.MRE11 and EXO1 nucleases degrade reversed forks and elicit MUS81-dependent fork rescue in BRCA2-deficient cells.Nat Commun2017;8:860 PMCID:PMC5643552

[16] Datta A,Sommers JA.WRN helicase safeguards deprotected replication forks in BRCA2-mutated cancer cells.Nat Commun2021;12:6561 PMCID:PMC8590011

[17] Sidorova J.A game of substrates: replication fork remodeling and its roles in genome stability and chemo-resistance.Cell Stress2017;1:115-33 PMCID:PMC5771654

[18] Berti M,Thangavel S.Human RECQ1 promotes restart of replication forks reversed by DNA topoisomerase I inhibition.Nat Struct Mol Biol2013;20:347-54 PMCID:PMC3897332

[19] Iannascoli C,Murfuni I,Pichierri P.The WRN exonuclease domain protects nascent strands from pathological MRE11/EXO1-dependent degradation.Nucleic Acids Res2015;43:9788-803 PMCID:PMC4787784

[20] Su F,Yang Y.Nonenzymatic role for WRN in preserving nascent DNA strands after replication stress.Cell Rep2014;9:1387-401 PMCID:PMC4782925

[21] Newman JA.RecQ helicases in DNA repair and cancer targets.Essays Biochem2020;64:819-30 PMCID:PMC7588665

[22] Karmakar P,Ramsden DA.Ku heterodimer binds to both ends of the Werner protein and functional interaction occurs at the Werner N-terminus.Nucleic Acids Res2002;30:3583-91 PMCID:PMC134248

[23] Grundy GJ,Arribas-Bosacoma R.The Ku-binding motif is a conserved module for recruitment and stimulation of non-homologous end-joining proteins.Nat Commun2016;7:11242 PMCID:PMC4831024

[24] Kusumoto-Matsuo R,Karmakar P,Ramsden D.Serines 440 and 467 in the Werner syndrome protein are phosphorylated by DNA-PK and affects its dynamics in response to DNA double strand breaks.Aging (Albany NY)2014;6:70-81 PMCID:PMC3927811

[25] Brosh RM Jr.Human premature aging, DNA repair and RecQ helicases.Nucleic Acids Res2007;35:7527-44 PMCID:PMC2190726

[26] Brosh RM Jr.DNA helicases involved in DNA repair and their roles in cancer.Nat Rev Cancer2013;13:542-58 PMCID:PMC4538698

[27] Shamanna RA,de Freitas JK,Croteau DL.WRN regulates pathway choice between classical and alternative non-homologous end joining.Nat Commun2016;7:13785 PMCID:PMC5150655

[28] Palermo V,Sanchez M.CDK1 phosphorylates WRN at collapsed replication forks.Nat Commun2016;7:12880 PMCID:PMC5028418

[29] Nimonkar AV,Kinoshita E.BLM-DNA2-RPA-MRN and EXO1-BLM-RPA-MRN constitute two DNA end resection machineries for human DNA break repair.Genes Dev2011;25:350-62 PMCID:PMC3042158

[30] Sturzenegger A,Kanagaraj R.DNA2 cooperates with the WRN and BLM RecQ helicases to mediate long-range DNA end resection in human cells.J Biol Chem2014;289:27314-26 PMCID:PMC4175362

[31] Lee JH,Kulikowicz T.CDK2 phosphorylation of Werner protein (WRN) contributes to WRN’s DNA double-strand break repair pathway choice.Aging Cell2021;20:e13484 PMCID:PMC8590104

[32] Gupta P,Chattopadhyay S.Pharmacological targeting of differential DNA repair, radio-sensitizes WRN-deficient cancer cells in vitro and in vivo.Biochem Pharmacol2021;186:114450

[33] Chu B,Paukstelis PJ.A DNA G-quadruplex/i-motif hybrid.Nucleic Acids Res2019;47:11921-30 PMCID:PMC7145706

[34] Kamath-Loeb AS,Schmitt MW.The Werner syndrome exonuclease facilitates DNA degradation and high fidelity DNA polymerization by human DNA polymerase δ.J Biol Chem2012;287:12480-90 PMCID:PMC3320997

[35] Shah SN,Meng X,Eckert KA.DNA structure and the Werner protein modulate human DNA polymerase delta-dependent replication dynamics within the common fragile site FRA16D.Nucleic Acids Res2010;38:1149-62 PMCID:PMC2831333

[36] Chan NL,Zhang T.The Werner syndrome protein promotes CAG/CTG repeat stability by resolving large (CAG)(n)/(CTG)(n) hairpins.J Biol Chem2012;287:30151-6 PMCID:PMC3436269

[37] Baird DM,Rowson J,Kipling D.Normal telomere erosion rates at the single cell level in Werner syndrome fibroblast cells.Hum Mol Genet2004;13:1515-24

[38] Sun L,Teng Y.WRN is recruited to damaged telomeres via its RQC domain and tankyrase1-mediated poly-ADP-ribosylation of TRF1.Nucleic Acids Res2017;45:3844-59 PMCID:PMC5397154

[39] Diotti R.Shelterin complex and associated factors at human telomeres.Nucleus2011;2:119-35 PMCID:PMC3127094

[40] Opresko PL,Podell ER.POT1 stimulates RecQ helicases WRN and BLM to unwind telomeric DNA substrates.J Biol Chem2005;280:32069-80

[41] Machwe A,Orren DK.TRF2 recruits the Werner syndrome (WRN) exonuclease for processing of telomeric DNA.Oncogene2004;23:149-56

[42] Edwards DN,Machwe A.Strand exchange of telomeric DNA catalyzed by the Werner syndrome protein (WRN) is specifically stimulated by TRF2.Nucleic Acids Res2014;42:7748-61 PMCID:PMC4081078

[43] Liu B,Yang X.MDM2-mediated degradation of WRN promotes cellular senescence in a p53-independent manner.Oncogene2019;38:2501-15

[44] Li M,Yi J.MIB1-mediated degradation of WRN promotes cellular senescence in response to camptothecin treatment.FASEB J2020;34:11488-97

[45] Patro BS,Bohr VA.WRN helicase regulates the ATR-CHK1-induced S-phase checkpoint pathway in response to topoisomerase-I-DNA covalent complexes.J Cell Sci2011;124:3967-79 PMCID:PMC3244981

[46] Cheng WH,Muftuoglu M.WRN is required for ATM activation and the S-phase checkpoint in response to interstrand cross-link-induced DNA double-strand breaks.Mol Biol Cell2008;19:3923-33 PMCID:PMC2526706

[47] Marabitti V,Malacaria E.ATM pathway activation limits R-loop-associated genomic instability in Werner syndrome cells.Nucleic Acids Res2019;47:3485-502 PMCID:PMC6468170

[48] Balajee AS,May A.The Werner syndrome protein is involved in RNA polymerase II transcription.Mol Biol Cell1999;10:2655-68 PMCID:PMC25497

[49] Shiratori M,Itoh C,Furuichi Y.WRN helicase accelerates the transcription of ribosomal RNA as a component of an RNA polymerase I-associated complex.Oncogene2002;21:2447-54

[50] Fang EF,Lautrup S.NAD⁺ augmentation restores mitophagy and limits accelerated aging in Werner syndrome.Nat Commun2019;10:5284 PMCID:PMC6872719

[51] Massip L,Paquet ER.Vitamin C restores healthy aging in a mouse model for Werner syndrome.FASEB J2010;24:158-72 PMCID:PMC3712979

[52] Maity J,Bohr VA.Acidic domain of WRNp is critical for autophagy and up-regulates age associated proteins.DNA Repair (Amst)2018;68:1-11 PMCID:PMC6338341

[53] Bellare G, Saha B, Patro BS. Targeting autophagy reverses de novo resistance in homologous recombination repair proficient breast cancers to PARP inhibition.Br J Cancer2021;124:1260-74 PMCID:PMC8007595

[54] Pai BG,Chakraborty S,Patro BS.Cross-talk between DNA damage and autophagy and its implication in cancer therapy. In: Bhutia SK, editors. Autophagy in tumor and tumor microenvironment. Singapore: Springer; 2020.

[55] Behan FM,Picco G.Prioritization of cancer therapeutic targets using CRISPR-Cas9 screens.Nature2019;568:511-6

[56] Chan EM,McFarland JM.WRN helicase is a synthetic lethal target in microsatellite unstable cancers.Nature2019;568:551-6 PMCID:PMC6580861

[57] Kategaya L,Hager JH.Werner syndrome helicase is required for the survival of cancer cells with microsatellite instability.iScience2019;13:488-97 PMCID:PMC6441948

[58] Lieb S,Kamper E.Werner syndrome helicase is a selective vulnerability of microsatellite instability-high tumor cells.Elife2019;8:e43333 PMCID:PMC6435321

[59] McDonald ER 3rd,Schlabach MR.Project DRIVE: a compendium of cancer dependencies and synthetic lethal relationships uncovered by large-scale, deep RNAi screening.Cell2017;170:577-92.e10

[60] Cowley GS,Vazquez F.Parallel genome-scale loss of function screens in 216 cancer cell lines for the identification of context-specific genetic dependencies.Sci Data2014;1:140035 PMCID:PMC4432652

[61] van Wietmarschen N,Nathan WJ.Repeat expansions confer WRN dependence in microsatellite-unstable cancers.Nature2020;586:292-8 PMCID:PMC8916167

[62] Picco G,van Vliet EJ.Cell Model Network UK GroupWerner helicase is a synthetic-lethal vulnerability in mismatch repair-deficient colorectal cancer refractory to targeted therapies, chemotherapy, and immunotherapy.Cancer Discov2021;11:1923-37

[63] Franchitto A,Prosperi E,Bignami M.Replication fork stalling in WRN-deficient cells is overcome by prompt activation of a MUS81-dependent pathway.J Cell Biol2008;183:241-52 PMCID:PMC2568021

[64] Aggarwal M,Shoemaker RH.Inhibition of helicase activity by a small molecule impairs Werner syndrome helicase (WRN) function in the cellular response to DNA damage or replication stress.Proc Natl Acad Sci U S A2011;108:1525-30 PMCID:PMC3029756

[65] Moles R,Chaib-Mezrag H.WRN-targeted therapy using inhibitors NSC 19630 and NSC 617145 induce apoptosis in HTLV-1-transformed adult T-cell leukemia cells.J Hematol Oncol2016;9:121 PMCID:PMC5103433

[66] Futami K,Shimamoto A,Furuichi Y.Increased chemotherapeutic activity of camptothecin in cancer cells by siRNA-induced silencing of WRN helicase.Biol Pharm Bull2007;30:1958-61

[67] Bou-Hanna C,Lode L.Acute cytotoxicity of MIRA-1/NSC19630, a mutant p53-reactivating small molecule, against human normal and cancer cells via a caspase-9-dependent apoptosis.Cancer Lett2015;359:211-7

[68] Aggarwal M,Sommers JA.Targeting an Achilles’ heel of cancer with a WRN helicase inhibitor.Cell Cycle2013;12:3329-35 PMCID:PMC3885643

[69] Aggarwal M,Sommers JA.Werner syndrome helicase has a critical role in DNA damage responses in the absence of a functional fanconi anemia pathway.Cancer Res2013;73:5497-507 PMCID:PMC3766423