過酸化水素処理後のゲノム安定性維持に関わるCST複合体の機能解析

Anderson BH, Kasher PR, Mayer J, Szynkiewicz M, Jenkinson EM, Bhaskar SS, Urquhart JE,

Daly SB, Dickerson JE, O'Sullivan J, Leibundgut EO, Muter J, Abdel-Salem GM, Babul-Hirji

R, Baxter P, Berger A, Bonafé L, Brunstom-Hernandez JE, Buckard JA, Chitayat D, Chong

WK, Cordelli DM, Ferreira P, Fluss J, Forrest EH, Franzoni E, Garone C, Hammans SR, Houge

G, Hughes I, Jacquemont S, Jeannet PY, Jefferson RJ, Kumar R, Kutschke G, Lundberg S,

Lourenço CM, Mehta R, Naidu S, Nischal KK, Nunes L, Ounap K, Philippart M, Prabhakar P,

Risen SR, Schiffmann R, Soh C, Stephenson JB, Stewart H, Stone J, Tolmie JL, van der Knaap

MS, Vieira JP, Vilain CN, Wakeling EL, Wermenbol V, Whitney A, Lovell SC, Meyer S,

Livingston JH, Baerlocher GM, Black GC, Rice GI, Crow YJ. Mutations in CTC1, encoding

conserved telomere maintenance component 1, cause Coats plus. Nat Genet. 2012 Jan

22;44(3):338-42. https://doi.org/10.1038/ng.1084 PMID: 22267198.

Barazas M, Annunziato S, Pettitt SJ, de Krijger I, Ghezraoui H, Roobol SJ, Lutz C, Frankum J,

Song FF, Brough R, Evers B, Gogola E, Bhin J, van de Ven M, van Gent DC, Jacobs JJL,

Chapman R, Lord CJ, Jonkers J, Rottenberg S. The CST Complex Mediates End Protection at

Double-Strand Breaks and Promotes PARP Inhibitor Sensitivity in BRCA1-Deficient Cells. Cell

Rep. 2018 May 15;23(7):2107-2118. https://doi.org/10.1016/j.celrep.2018.04.046 PMID:

29768208.

Berniak K, Rybak P, Bernas T, Zarębski M, Biela E, Zhao H, Darzynkiewicz Z, Dobrucki JW.

Relationship between DNA damage response, initiated by camptothecin or oxidative stress, and

DNA replication, analyzed by quantitative 3D image analysis. Cytometry A. 2013

Oct;83(10):913-24. https://doi.org/10.1002/cyto.a.22327 PMID: 23846844.

Bhat KP, Cortez D. RPA and RAD51: fork reversal, fork protection, and genome stability. Nat

Struct Mol Biol. 2018 Jun;25(6):446-453. https://doi.org/10.1038/s41594-018-0075-z PMID:

29807999.

Bhattacharjee A, Stewart J, Chaiken M, Price CM. STN1 OB Fold Mutation Alters DNA

Binding and Affects Selective Aspects of CST Function. PLoS Genet. 2016 Sep

30;12(9):e1006342. https://doi.org/10.1371/journal.pgen.1006342 PMID: 27690379.

Blackford AN, Jackson SP. ATM, ATR, and DNA-PK: The Trinity at the Heart of the DNA

Damage Response. Mol Cell. 2017 Jun 15;66(6):801-817.

98

https://doi.org/10.1016/j.molcel.2017.05.015 PMID: 28622525.

Bonilla B, Hengel SR, Grundy MK, Bernstein KA. RAD51 Gene Family Structure and

Function. Annu Rev Genet. 2020 Nov 23;54:25-46. https://doi.org/10.1146/annurev-genet021920-092410 PMID: 32663049.

Cai SW, Zinder JC, Svetlov V, Bush MW, Nudler E, Walz T, de Lange T. Cryo-EM structure of

the human CST-Polα/primase complex in a recruitment state. Nat Struct Mol Biol. 2022

Aug;29(8):813-819. https://doi.org/10.1038/s41594-022-00766-yPMID: 35578024.

Casteel DE, Zhuang S, Zeng Y, Perrino FW, Boss GR, Goulian M, Pilz RB. A DNA polymerasea・primase cofactor with homology to replication protein A-32 regulates DNA replication in

mammalian cells. J Biol Chem. 2009 Feb 27;284(9):5807-18.

https://doi.org/10.1074/jbc.M807593200 PMID: 19119139.

Chastain M, Zhou Q, Shiva O, Fadri-Moskwik M, Whitmore L, Jia P, Dai X, Huang C, Ye P,

Chai W. Human CST Facilitates Genome-wide RAD51 Recruitment to GC-Rich Repetitive

Sequences in Response to Replication Stress. Cell Rep. 2016 Aug 2;16(5):1300-1314.

https://doi.org/10.1016/j.celrep.2016.06.077 Erratum in: Cell Rep. 2016 Aug 16;16(7):2048.

PMID: 27487043.

Chen LY, Redon S, Lingner J. The human CST complex is a terminator of telomerase activity.

Nature. 2012 Aug 23;488(7412):540-4. https://doi.org/10.1038/nature11269 PMID: 22763445.

Chen Q, Bian C, Wang X, Liu X, Ahmad Kassab M, Yu Y, Yu X. ADP-ribosylation of histone

variant H2AX promotes base excision repair. EMBO J. 2021 Jan 15;40(2):e104542.

https://doi.org/10.15252/embj.2020104542 PMID: 33264433.

Cong K, Peng M, Kousholt AN, Lee WTC, Lee S, Nayak S, Krais J, VanderVere-Carozza PS,

Pawelczak KS, Calvo J, Panzarino NJ, Turchi JJ, Johnson N, Jonkers J, Rothenberg E, Cantor

SB. Replication gaps are a key determinant of PARP inhibitor synthetic lethality with BRCA

deficiency. Mol Cell. 2021 Aug 5;81(15):3128-3144.e7.

https://doi.org/10.1016/j.molcel.2021.06.011 Erratum in: Mol Cell. 2021 Aug 5;81(15):3227.

PMID: 34216544.

de Feraudy S, Limoli CL, Giedzinski E, Karentz D, Marti TM, Feeney L, Cleaver JE. Pol eta is

99

required for DNA replication during nucleotide deprivation by hydroxyurea. Oncogene. 2007

Aug 23;26(39):5713-21. https://doi.org/10.1038/sj.onc.1210385 PMID: 17369853.

de Lange T. Shelterin: the protein complex that shapes and safeguards human telomeres. Genes

Dev. 2005 Sep 15;19(18):2100-10. https://doi.org/10.1101/gad.1346005 PMID: 16166375.

Doksani Y, de Lange T. The role of double-strand break repair pathways at functional and

dysfunctional telomeres. Cold Spring Harb Perspect Biol. 2014 Sep 16;6(12):a016576.

https://doi.org/10.1101/cshperspect.a016576 PMID: 25228584.

Driessens N, Versteyhe S, Ghaddhab C, Burniat A, De Deken X, Van Sande J, Dumont JE, Miot

F, Corvilain B. Hydrogen peroxide induces DNA single- and double-strand breaks in thyroid

cells and is therefore a potential mutagen for this organ. Endocr Relat Cancer. 2009

Sep;16(3):845-56. https://doi.org/10.1677/ERC-09-0020 PMID: 19509065.

Elvers I, Johansson F, Groth P, Erixon K, Helleday T. UV stalled replication forks restart by repriming in human fibroblasts. Nucleic Acids Res. 2011 Sep 1;39(16):7049-57.

https://doi.org/10.1093/nar/gkr420. PMID: 21646340.

Feng X, Hsu SJ, Bhattacharjee A, Wang Y, Diao J, Price CM. CTC1-STN1 terminates

telomerase while STN1-TEN1 enables C-strand synthesis during telomere replication in colon

cancer cells. Nat Commun. 2018 Jul 19;9(1):2827. https://doi.org/10.1038/s41467-018-05154-z

PMID: 30026550.

Gaillard H, García-Muse T, Aguilera A. Replication stress and cancer. Nat Rev Cancer. 2015

May;15(5):276-89. https://doi.org/10.1038/nrc3916 PMID: 25907220.

Ganduri S, Lue NF. STN1-POLA2 interaction provides a basis for primase-pol α stimulation by

human STN1. Nucleic Acids Res. 2017 Sep 19;45(16):9455-9466.

https://doi.org/10.1093/nar/gkx621 PMID: 28934486.

Garvik B, Carson M, Hartwell L. Single-stranded DNA arising at telomeres in cdc13 mutants

may constitute a specific signal for the RAD9 checkpoint. Mol Cell Biol. 1995

Nov;15(11):6128-38. https://doi.org/10.1128/MCB.15.11.6128 Erratum in: Mol Cell Biol 1996

Jan;16(1):457. PMID: 7565765.

100

Goulian M, Heard CJ. The mechanism of action of an accessory protein for DNA polymerase

alpha/primase. J Biol Chem. 1990 Aug 5;265(22):13231-9. Erratum in: J Biol Chem 1990 Nov

5;265(31):19369. PMID: 2376593.

Goulian M, Heard CJ, Grimm SL. Purification and properties of an accessory protein for DNA

polymerase alpha/primase. J Biol Chem. 1990 Aug 5;265(22):13221-30. PMID: 2165497.

Grandin N, Damon C, Charbonneau M. Ten1 functions in telomere end protection and length

regulation in association with Stn1 and Cdc13. EMBO J. 2001 Mar 1;20(5):1173-83.

https://doi.org/10.1093/emboj/20.5.1173 PMID: 11230140.

Grandin N, Reed SI, Charbonneau M. Stn1, a new Saccharomyces cerevisiae protein, is

implicated in telomere size regulation in association with Cdc13. Genes Dev. 1997 Feb

15;11(4):512-27. https://doi.org/10.1101/gad.11.4.512 PMID: 9042864.

Griffith JD, Comeau L, Rosenfield S, Stansel RM, Bianchi A, Moss H, de Lange T. Mammalian

telomeres end in a large duplex loop. Cell. 1999 May 14;97(4):503-14.

https://doi.org/10.1016/s0092-8674(00)80760-6 PMID: 10338214.

Groth P, Ausländer S, Majumder MM, Schultz N, Johansson F, Petermann E, Helleday T.

Methylated DNA causes a physical block to replication forks independently of damage

signalling, O(6)-methylguanine or DNA single-strand breaks and results in DNA damage. J Mol

Biol. 2010 Sep 10;402(1):70-82. https://doi.org/10.1016/j.jmb.2010.07.010 PMID: 20643142.

Gu P, Chang S. Functional characterization of human CTC1 mutations reveals novel

mechanisms responsible for the pathogenesis of the telomere disease Coats plus. Aging Cell.

2013 Dec;12(6):1100-9. https://doi.org/10.1111/acel.12139. Epub 2013 Sep 4. PMID:

23869908.

Gyori BM, Venkatachalam G, Thiagarajan PS, Hsu D, Clement MV. OpenComet: an automated

tool for comet assay image analysis. Redox Biol. 2014 Jan 9;2:457-65.

https://doi.org/10.1016/j.redox.2013.12.020 Erratum in: Redox Biol. 2021 Apr;40:101876.

PMID: 24624335.

Hanada K, Budzowska M, Davies SL, van Drunen E, Onizawa H, Beverloo HB, Maas A, Essers

J, Hickson ID, Kanaar R. The structure-specific endonuclease Mus81 contributes to replication

101

restart by generating double-strand DNA breaks. Nat Struct Mol Biol. 2007 Nov;14(11):1096104. https://doi.org/10.1038/nsmb1313 PMID: 17934473.

He Q, Lin X, Chavez BL, Agrawal S, Lusk BL, Lim CJ. Structures of the human CST-Polαprimase complex bound to telomere templates. Nature. 2022 Aug;608(7924):826-832.

https://doi.org/10.1038/s41586-022-05040-1 PMID: 35830881.

Hom RA, Wuttke DS. Human CST Prefers G-Rich but Not Necessarily Telomeric Sequences.

Biochemistry. 2017 Aug 15;56(32):4210-4218. https://doi.org/10.1021/acs.biochem.7b00584

PMID: 28726394.

Hsiang YH, Lihou MG, Liu LF. Arrest of replication forks by drug-stabilized topoisomerase IDNA cleavable complexes as a mechanism of cell killing by camptothecin. Cancer Res. 1989

Sep 15;49(18):5077-82. PMID: 2548710.

Huang C, Dai X, Chai W. Human Stn1 protects telomere integrity by promoting efficient

lagging-strand synthesis at telomeres and mediating C-strand fill-in. Cell Res. 2012

Dec;22(12):1681-95. https://doi.org/10.1038/cr.2012.132 PMID: 22964711.

Huang F, Mazina OM, Zentner IJ, Cocklin S, Mazin AV. Inhibition of homologous

recombination in human cells by targeting RAD51 recombinase. J Med Chem. 2012 Apr

12;55(7):3011-20. https://doi.org/10.1021/jm201173g PMID: 22380680.

Huang F, Motlekar NA, Burgwin CM, Napper AD, Diamond SL, Mazin AV. Identification of

specific inhibitors of human RAD51 recombinase using high-throughput screening. ACS Chem

Biol. 2011 Jun 17;6(6):628-35. https://doi.org/10.1021/cb100428c PMID: 21428443.

Inano S, Sato K, Katsuki Y, Kobayashi W, Tanaka H, Nakajima K, Nakada S, Miyoshi H, Knies

K, Takaori-Kondo A, Schindler D, Ishiai M, Kurumizaka H, Takata M. RFWD3-Mediated

Ubiquitination Promotes Timely Removal of Both RPA and RAD51 from DNA Damage Sites to

Facilitate Homologous Recombination. Mol Cell. 2020 Apr 2;78(1):192.

https://doi.org/10.1016/j.molcel.2020.03.026 Erratum for: Mol Cell. 2017 Jun 1;66(5):622634.e8. PMID: 32243829.

Karnani N, Dutta A. The effect of the intra-S-phase checkpoint on origins of replication in

human cells. Genes Dev. 2011 Mar 15;25(6):621-33. https://doi.org/10.1101/gad.2029711

102

PMID: 21406556.

Kasbek C, Wang F, Price CM. Human TEN1 maintains telomere integrity and functions in

genome-wide replication restart. J Biol Chem. 2013 Oct 18;288(42):30139-30150.

https://doi.org/10.1074/jbc.M113.493478 PMID: 24025336.

Katsube T, Mori M, Tsuji H, Shiomi T, Wang B, Liu Q, Nenoi M, Onoda M. Most hydrogen

peroxide-induced histone H2AX phosphorylation is mediated by ATR and is not dependent on

DNA double-strand breaks. J Biochem. 2014 Aug;156(2):85-95.

https://doi.org/10.1093/jb/mvu021 PMID: 24682951.

Kopp B, Khoury L, Audebert M. Validation of the γH2AX biomarker for genotoxicity

assessment: a review. Arch Toxicol. 2019 Aug;93(8):2103-2114. https://doi.org/10.1007/s00204019-02511-9 PMID: 31289893.

Krokan HE, Bjørås M. Base excision repair. Cold Spring Harb Perspect Biol. 2013 Apr

1;5(4):a012583. https://doi.org/10.1101/cshperspect.a012583 PMID: 23545420.

Kurose A, Tanaka T, Huang X, Traganos F, Dai W, Darzynkiewicz Z. Effects of hydroxyurea

and aphidicolin on phosphorylation of ataxia telangiectasia mutated on Ser 1981 and histone

H2AX on Ser 139 in relation to cell cycle phase and induction of apoptosis. Cytometry A. 2006

Apr;69(4):212-21. https://doi.org/10.1002/cyto.a.20241 PMID: 16528719.

Lei KH, Yang HL, Chang HY, Yeh HY, Nguyen DD, Lee TY, Lyu X, Chastain M, Chai W, Li

HW, Chi P. Crosstalk between CST and RPA regulates RAD51 activity during replication stress.

Nat Commun. 2021 Nov 5;12(1):6412. https://doi.org/10.1038/s41467-021-26624-x PMID:

34741010.

Lim CJ, Barbour AT, Zaug AJ, Goodrich KJ, McKay AE, Wuttke DS, Cech TR. The structure of

human CST reveals a decameric assembly bound to telomeric DNA. Science. 2020 Jun

5;368(6495):1081-1085. https://doi.org/10.1126/science.aaz9649 PMID: 32499435.

Lim CJ, Cech TR. Shaping human telomeres: from shelterin and CST complexes to telomeric

chromatin organization. Nat Rev Mol Cell Biol. 2021 Apr;22(4):283-298.

https://doi.org/10.1038/s41580-021-00328-y Erratum in: Nat Rev Mol Cell Biol. 2021 Feb 19;:

PMID: 33564154.

103

Limsirichaikul S, Niimi A, Fawcett H, Lehmann A, Yamashita S, Ogi T. A rapid non-radioactive

technique for measurement of repair synthesis in primary human fibroblasts by incorporation of

ethynyl deoxyuridine (EdU). Nucleic Acids Res. 2009 Mar;37(4):e31.

https://doi.org/10.1093/nar/gkp023 PMID: 19179371.

Lowndes NF, Toh GW. DNA repair: the importance of phosphorylating histone H2AX. Curr

Biol. 2005 Feb 8;15(3):R99-R102. https://doi.org/10.1016/j.cub.2005.01.029 PMID: 15694301.

Lue NF, Chan J, Wright WE, Hurwitz J. The CDC13-STN1-TEN1 complex stimulates Pol α

activity by promoting RNA priming and primase-to-polymerase switch. Nat Commun. 2014

Dec 12;5:5762. https://doi.org/10.1038/ncomms6762 PMID: 25503194.

Lue NF, Zhou R, Chico L, Mao N, Steinberg-Neifach O, Ha T. The telomere capping complex

CST has an unusual stoichiometry, makes multipartite interaction with G-Tails, and unfolds

higher-order G-tail structures. PLoS Genet. 2013;9(1):e1003145.

https://doi.org/10.1371/journal.pgen.1003145 PMID: 23300477

Lyu X, Lei KH, Biak Sang P, Shiva O, Chastain M, Chi P, Chai W. Human CST complex

protects stalled replication forks by directly blocking MRE11 degradation of nascent-strand

DNA. EMBO J. 2021 Jan 15;40(2):e103654. https://doi.org/10.15252/embj.2019103654 PMID:

33210317.

Mirman Z, Sasi NK, King A, Chapman JR, de Lange T. 53BP1-shieldin-dependent DSB

processing in BRCA1-deficient cells requires CST-Polα-primase fill-in synthesis. Nat Cell Biol.

2022 Jan;24(1):51-61. https://doi.org/10.1038/s41556-021-00812-9 PMID: 35027730.

Miyake Y, Nakamura M, Nabetani A, Shimamura S, Tamura M, Yonehara S, Saito M, Ishikawa

F. RPA-like mammalian Ctc1-Stn1-Ten1 complex binds to single-stranded DNA and protects

telomeres independently of the Pot1 pathway. Mol Cell. 2009 Oct 23;36(2):193-206.

https://doi.org/10.1016/j.molcel.2009.08.009 PMID: 19854130.

Murzin AG. OB(oligonucleotide/oligosaccharide binding)-fold: common structural and

functional solution for non-homologous sequences. EMBO J. 1993 Mar;12(3):861-7.

https://doi.org/10.1002/j.1460-2075.1993.tb05726.x PMID: 8458342.

104

Nakaoka H, Nishiyama A, Saito M, Ishikawa F. Xenopus laevis Ctc1-Stn1-Ten1 (xCST) protein

complex is involved in priming DNA synthesis on single-stranded DNA template in Xenopus

egg extract. J Biol Chem. 2012 Jan 2;287(1):619-627. https://doi.org/10.1074/jbc.M111.263723

PMID: 22086929.

Nguyen BC, Tawata S. The Chemistry and Biological Activities of Mimosine: A Review.

Phytother Res. 2016 Aug;30(8):1230-42. https://doi.org/10.1002/ptr.5636 PMID: 27213712.

Nguyen DD, Kim E, Le NT, Ding X, Jaiswal RK, Kostlan RJ, Nguyen TNT, Shiva O, Le MT,

Chai W. Deficiency in mammalian STN1 promotes colon cancer development via inhibiting

DNA repair. Sci Adv. 2023 May 10;9(19):eadd8023. https://doi.org/10.1126/sciadv.add8023

PMID: 37163605.

Novo E, Parola M. Redox mechanisms in hepatic chronic wound healing and fibrogenesis.

Fibrogenesis Tissue Repair. 2008 Oct 13;1(1):5. https://doi.org/10.1186/1755-1536-1-5 PMID:

19014652.

Olovnikov AM. A theory of marginotomy. The incomplete copying of template margin in

enzymic synthesis of polynucleotides and biological significance of the phenomenon. J Theor

Biol. 1973 Sep 14;41(1):181-90. https://doi.org/10.1016/0022-5193(73)90198-7 PMID:

4754905.

Petermann E, Orta ML, Issaeva N, Schultz N, Helleday T. Hydroxyurea-stalled replication forks

become progressively inactivated and require two different RAD51-mediated pathways for

restart and repair. Mol Cell. 2010 Feb 26;37(4):492-502.

https://doi.org/10.1016/j.molcel.2010.01.021 PMID: 20188668.

Poetsch AR. The genomics of oxidative DNA damage, repair, and resulting mutagenesis.

Comput Struct Biotechnol J. 2020 Jan 7;18:207-219. https://doi.org/10.1016/j.csbj.2019.12.013

PMID: 31993111.

Primo LMF, Teixeira LK. DNA replication stress: oncogenes in the spotlight. Genet Mol Biol.

2019 Dec 13;43(1 suppl 1):e20190138. https://doi.org/10.1590/1678-4685GMB-2019-0138

PMID: 31930281.

Regairaz M, Zhang YW, Fu H, Agama KK, Tata N, Agrawal S, Aladjem MI, Pommier Y.

105

Mus81-mediated DNA cleavage resolves replication forks stalled by topoisomerase I-DNA

complexes. J Cell Biol. 2011 Nov 28;195(5):739-49. https://doi.org/10.1083/jcb.201104003

PMID: 22123861.

Renkawitz J, Lademann CA, Jentsch S. Mechanisms and principles of homology search during

recombination. Nat Rev Mol Cell Biol. 2014 Jun;15(6):369-83. https://doi.org/10.1038/nrm3805

PMID: 24824069.

Sakaguchi K, Ishibashi T, Uchiyama Y, Iwabata K. The multi-replication protein A (RPA)

system--a new perspective. FEBS J. 2009 Feb;276(4):943-63. https://doi.org/10.1111/j.17424658.2008.06841.x. PMID: 19154342.

Sarek G, Vannier JB, Panier S, Petrini JHJ, Boulton SJ. TRF2 recruits RTEL1 to telomeres in S

phase to promote t-loop unwinding. Mol Cell. 2015 Feb 19;57(4):622-635.

https://doi.org/10.1016/j.molcel.2014.12.024 Erratum in: Mol Cell. 2016 Mar 3;61(5):788-9.

Erratum in: Mol Cell. 2016 Mar 3;61(5):788-789. PMID: 25620558.

Schindelin J, Arganda-Carreras I, Frise E, Kaynig V, Longair M, Pietzsch T, Preibisch S,

Rueden C, Saalfeld S, Schmid B, Tinevez JY, White DJ, Hartenstein V, Eliceiri K, Tomancak P,

Cardona A. Fiji: an open-source platform for biological-image analysis. Nat Methods. 2012 Jun

28;9(7):676-82. https://doi.org/10.1038/nmeth.2019. PMID: 22743772.

Scully R, Panday A, Elango R, Willis NA. DNA double-strand break repair-pathway choice in

somatic mammalian cells. Nat Rev Mol Cell Biol. 2019 Nov;20(11):698-714.

https://doi.org/10.1038/s41580-019-0152-0 PMID: 31263220.

Simon AJ, Lev A, Zhang Y, Weiss B, Rylova A, Eyal E, Kol N, Barel O, Cesarkas K, Soudack

M, Greenberg-Kushnir N, Rhodes M, Wiest DL, Schiby G, Barshack I, Katz S, Pras E, Poran H,

Reznik-Wolf H, Ribakovsky E, Simon C, Hazou W, Sidi Y, Lahad A, Katzir H, Sagie S, Aqeilan

HA, Glousker G, Amariglio N, Tzfati Y, Selig S, Rechavi G, Somech R. Mutations in STN1

cause Coats plus syndrome and are associated with genomic and telomere defects. J Exp Med.

2016 Jul 25;213(8):1429-40. https://doi.org/10.1084/jem.20151618 PMID: 27432940.

Srinivas US, Tan BWQ, Vellayappan BA, Jeyasekharan AD. ROS and the DNA damage

response in cancer. Redox Biol. 2019 Jul;25:101084.

https://doi.org/10.1016/j.redox.2018.101084 PMID: 30612957.

106

Stewart JA, Wang F, Chaiken MF, Kasbek C, Chastain PD 2nd, Wright WE, Price CM. Human

CST promotes telomere duplex replication and general replication restart after fork stalling.

EMBO J. 2012 Aug 29;31(17):3537-49. https://doi.org/10.1038/emboj.2012.215 PMID:

22863775.

Surovtseva YV, Churikov D, Boltz KA, Song X, Lamb JC, Warrington R, Leehy K, Heacock M,

Price CM, Shippen DE. Conserved telomere maintenance component 1 interacts with STN1 and

maintains chromosome ends in higher eukaryotes. Mol Cell. 2009 Oct 23;36(2):207-18.

https://doi.org/10.1016/j.molcel.2009.09.017 PMID: 19854131.

Tanaka T, Halicka D, Traganos F, Darzynkiewicz Z. Cytometric analysis of DNA damage:

phosphorylation of histone H2AX as a marker of DNA double-strand breaks (DSBs). Methods

Mol Biol. 2009;523:161-8. https://doi.org/10.1007/978-1-59745-190-1_11 PMID: 19381940.

Vallerga MB, Mansilla SF, Federico MB, Bertolin AP, Gottifredi V. Rad51 recombinase

prevents Mre11 nuclease-dependent degradation and excessive PrimPol-mediated elongation of

nascent DNA after UV irradiation. Proc Natl Acad Sci U S A. 2015 Dec 1;112(48):E6624-33.

https://doi.org/10.1073/pnas.1508543112 PMID: 26627254.

Vaurs M, Naiman K, Bouabboune C, Rai S, Ptasińska K, Rives M, Matmati S, Carr AM, Géli V,

Coulon S. Stn1-Ten1 and Taz1 independently promote replication of subtelomeric fragile

sequences in fission yeast. Cell Rep. 2023 May 25;42(6):112537.

https://doi.org/10.1016/j.celrep.2023.112537 PMID: 37243596.

Vrtis KB, Dewar JM, Chistol G, Wu RA, Graham TGW, Walter JC. Single-strand DNA breaks

cause replisome disassembly. Mol Cell. 2021 Mar 18;81(6):1309-1318.e6.

https://doi.org/10.1016/j.molcel.2020.12.039 PMID: 33484638.

Wang F, Stewart JA, Kasbek C, Zhao Y, Wright WE, Price CM. Human CST has independent

functions during telomere duplex replication and C-strand fill-in. Cell Rep. 2012 Nov

29;2(5):1096-103. https://doi.org/10.1016/j.celrep.2012.10.007 PMID: 23142664.

Wang F, Stewart J, Price CM. Human CST abundance determines recovery from diverse forms

of DNA damage and replication stress. Cell Cycle. 2014;13(22):3488-98.

https://doi.org/10.4161/15384101.2014.964100 PMID: 25483097.

107

Wang Y, Chai W. Pathogenic CTC1 mutations cause global genome instabilities under

replication stress. Nucleic Acids Res. 2018 May 4;46(8):3981-3992.

https://doi.org/10.1093/nar/gky114 PMID: 29481669.

Ward IM, Chen J. Histone H2AX is phosphorylated in an ATR-dependent manner in response to

replicational stress. J Biol Chem. 2001 Dec 21;276(51):47759-62.

https://doi.org/10.1074/jbc.C100569200 PMID: 11673449.

Wassing IE, Esashi F. RAD51: Beyond the break. Semin Cell Dev Biol. 2021 May;113:38-46.

https://doi.org/10.1016/j.semcdb.2020.08.010 PMID: 32938550.

Watson JD. Origin of concatemeric T7 DNA. Nat New Biol. 1972 Oct 18;239(94):197-201.

https://doi.org/10.1038/newbio239197a0 PMID: 4507727.

Willis N, Rhind N. Regulation of DNA replication by the S-phase DNA damage checkpoint.

Cell Div. 2009 Jul 3;4:13. https://doi.org/10.1186/1747-1028-4-13 PMID: 19575778.

Wu P, Takai H, de Lange T. Telomeric 3' overhangs derive from resection by Exo1 and Apollo

and fill-in by POT1b-associated CST. Cell. 2012 Jul 6;150(1):39-52.

https://doi.org/10.1016/j.cell.2012.05.026 PMID: 22748632.

Wyatt MD, Pittman DL. Methylating agents and DNA repair responses: Methylated bases and

sources of strand breaks. Chem Res Toxicol. 2006 Dec;19(12):1580-94.

https://doi.org/10.1021/tx060164e PMID: 17173371.

Yamamoto I, Nakaoka H, Takikawa M, Tashiro S, Kanoh J, Miyoshi T, Ishikawa F. Fission

yeast Stn1 maintains stability of repetitive DNA at subtelomere and ribosomal DNA regions.

Nucleic Acids Res. 2021 Oct 11;49(18):10465-10476. https://doi.org/10.1093/nar/gkab767

PMID: 34520548.

Zaug AJ, Goodrich KJ, Song JJ, Sullivan AE, Cech TR. Reconstitution of a telomeric replicon

organized by CST. Nature. 2022 Aug;608(7924):819-825. https://doi.org/10.1038/s41586-02204930-8 PMID: 35831508.

Zhang K, Zheng DQ, Sui Y, Qi L, Petes TD. Genome-wide analysis of genomic alterations

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induced by oxidative DNA damage in yeast. Nucleic Acids Res. 2019 Apr 23;47(7):3521-3535.

https://doi.org/10.1093/nar/gkz027 PMID: 30668788.

Zhou Q, Chai W. Suppression of STN1 enhances the cytotoxicity of chemotherapeutic agents in

cancer cells by elevating DNA damage. Oncol Lett. 2016 Aug;12(2):800-808.

https://doi.org/10.3892/ol.2016.4676 PMID: 27446354.

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謝辞

本研究を行うという大変貴重な機会を与えて下さり、大学院入院前より常に幅広い見識から

終始懇切なる御指導ならびに御鞭撻を賜りました、指導教員の石川冬木名誉教授に深く感謝

し、厚く御礼申し上げます。学位申請に際して主査となっていただいた上村匡教授に心より感

謝致します。実験の計画段階から学術論文公表に至るまでの研究全般に渡り懇切丁寧な御

指導を賜りました、中岡秀憲助教と理化学研究所の三好知一郎チームリーダーに深く感謝致

します。実験を行った際に、必要な実験プロトコルや機器類・ソフトウェアをご提供いただいた

高原和彦准教授と髙田穣特任教授に厚く御礼申し上げます。学術論文を投稿した際に、的

確な英文校正と学術的なアドバイスを下さった James Alan Hejna 特命教授に深く感謝致

します。 研究内容や進路に関する御助言を数多く与えて下さり、研究者としての道筋をお示し

いただいた京都府立大学の森川耿右博士に心より感謝致します。

本研究の起点となった実験データをもたらして下さった嶋祐輔氏に深く感謝の意を表します。

研究室離籍後も折に触れてお気遣いを下さり御助言いただいた林眞理客員准教授と東京理

科大学の定家真人准教授に心より感謝致します。共通試薬の作製等で研究を支えて下さった

技術専門職員の渡邉祐三氏、研究室における事務処理を一手に引き受けて下さると共に励

ましやお気遣いの言葉を常に掛けて下さった、秘書の白淵愛依子氏、木村恵子氏、林奈緒美

氏、津田貴子氏、平田陽子氏、ならびに阪本真弓氏に深く感謝の意を表します。

充実した研究生活を思う存分に行い、人間性・社会性・研究面で大きく成長した７年間を共

に過ごした石川研究室の皆様に深く感謝致します。

最後に、最大の理解者として研究生活を物心両面から応援して下さった家族に深く感謝し、

厚く御礼申し上げて謝辞にかえさせていただきます。

本学位論文は以下の学術論文の内容に基づいて書かれたものである。

Hara T, Nakaoka H, Miyoshi T, Ishikawa F. The CST complex facilitates cell survival under

oxidative genotoxic stress. PLoS One. 2023 Aug 17;18(8):e0289304.

https://doi.org/10.1371/journal.pone.0289304 PMID: 37590191.

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