Mcm10 deficiency causes defective-replisome-induced mutagenesis and a dependency on error-free postreplicative repair

Cell Cycle

Volumn 13, Issue 11, 2014, Pages 1737-1748

  Becker, Jordan R ^a   Nguyen, Hai Dang ^a,b   Wang, Xiaohan ^a   Bielinsky, Anja Katrin ^a  

^a UNIVERSITY OF MINNESOTA   (United States)

^b HARVARD MEDICAL SCHOOL   (United States)

Author keywords

DNA replication; DRIM; Mcm10; PCNA ubiquitination; Pol1; Sumoylation; Translesion synthesis

Indexed keywords

CYCLINE; DNA POLYMERASE; MCM10 PROTEIN; SCAFFOLD PROTEIN; UNCLASSIFIED DRUG; MCM10 PROTEIN, S CEREVISIAE; MINICHROMOSOME MAINTENANCE PROTEIN; SACCHAROMYCES CEREVISIAE PROTEIN;

ALLELE; ARTICLE; CELL VIABILITY; CONTROLLED STUDY; DEFECTIVE REPLISOME INDUCED MUTAGENESIS; DNA REPAIR; DNA REPLICATION; DNA SYNTHESIS; MUTAGENESIS; MUTANT; NONHUMAN; POSTREPLICATIVE REPAIR; PROTEIN DEFICIENCY; REPLISOME; TEMPERATURE SENSITIVITY; UBIQUITINATION; CELL CYCLE CHECKPOINT; DEFICIENCY; FLOW CYTOMETRY; GENE VECTOR; GENETICS; METABOLISM; MUTATION RATE; NONPARAMETRIC TEST; SACCHAROMYCES CEREVISIAE;

CELL CYCLE CHECKPOINTS; DNA REPAIR; DNA REPLICATION; FLOW CYTOMETRY; GENETIC VECTORS; MINICHROMOSOME MAINTENANCE PROTEINS; MUTAGENESIS; MUTATION RATE; PROLIFERATING CELL NUCLEAR ANTIGEN; SACCHAROMYCES CEREVISIAE; SACCHAROMYCES CEREVISIAE PROTEINS; STATISTICS, NONPARAMETRIC; UBIQUITINATION;

EID: 84901842871     PISSN: 15384101     EISSN: 15514005     Source Type: Journal    
DOI: 10.4161/cc.28652     Document Type: Article

References (79)

1. Hanahan D, Weinberg RA. Hallmarks of cancer: the next generation. Cell 2011; 144:646-74; PMID:21376230; http://dx.doi.org/10.1016/j.cell.2011.02.013
2. Myung K, Chen C, Kolodner RD. Multiple pathways cooperate in the suppression of genome instability in Saccharomyces cerevisiae. Nature 2001; 411:1073-6; PMID:11429610; http://dx.doi.org/10.1038/35082608
3. Moldovan GL, Pfander B, Jentsch S. PCNA, the maestro of the replication fork. Cell 2007; 129:665-79; PMID:17512402; http://dx.doi.org/10.1016/j.cell. 2007.05.003
4. Burgers PM. Polymerase dynamics at the eukaryotic DNA replication fork. J Biol Chem 2009; 284:4041-5; PMID:18835809; http://dx.doi.org/10.1074/jbc. R800062200
5. Lopes M, Foiani M, Sogo JM. Multiple mechanisms control chromosome integrity after replication fork uncoupling and restart at irreparable UV lesions. Mol Cell 2006; 21:15-27; PMID:16387650; http://dx.doi.org/10.1016/j. molcel.2005.11.015
6. Hoege C, Pfander B, Moldovan GL, Pyrowolakis G, Jentsch S. RAD6-dependent DNA repair is linked to modification of PCNA by ubiquitin and SUMO. Nature 2002; 419:135-41; PMID:12226657; http://dx.doi.org/10.1038/nature00991
7. Davies AA, Huttner D, Daigaku Y, Chen S, Ulrich HD. Activation of ubiquitin-dependent DNA damage bypass is mediated by replication protein a. Mol Cell 2008; 29:625-36; PMID:18342608; http://dx.doi.org/10.1016/j.molcel.2007.12. 016
8. Branzei D. Ubiquitin family modifications and template switching. FEBS Lett 2011; 585:2810-7; PMID:21539841; http://dx.doi.org/10.1016/j.febslet.2011. 04.053
9. Branzei D, Foiani M. Maintaining genome stability at the replication fork. Nat Rev Mol Cell Biol 2010; 11:208-19; PMID:20177396; http://dx.doi.org/ 10.1038/nrm2852
10. Prakash S, Johnson RE, Prakash L. Eukaryotic translesion synthesis DNA polymerases: specificity of structure and function. Annu Rev Biochem 2005; 74:317-53; PMID:15952890; http://dx.doi.org/10.1146/annurev.biochem.74.082803. 133250
11. Waters LS, Minesinger BK, Wiltrout ME, D'Souza S, Woodruff RV, Walker GC. Eukaryotic translesion polymerases and their roles and regulation in DNA damage tolerance. Microbiol Mol Biol Rev 2009; 73:134-54; PMID:19258535; http://dx.doi.org/10.1128/MMBR.00034-08
12. Northam MR, Garg P, Baitin DM, Burgers PM, Shcherbakova PV. A novel function of DNA polymerase ζ regulated by PCNA. EMBO J 2006; 25:4316-25; PMID:16957771; http://dx.doi.org/10.1038/sj.emboj.7601320
13. Northam MR, Robinson HA, Kochenova OV, Shcherbakova PV. Participation of DNA polymerase ζ in replication of undamaged DNA in Saccharomyces cerevisiae. Genetics 2010; 184:27-42; PMID:19841096; http://dx.doi.org/10.1534/ genetics.109.107482
14. Suzuki M, Niimi A, Limsirichaikul S, Tomida S, Miao Huang Q, Izuta S, Usukura J, Itoh Y, Hishida T, Akashi T, et al. PCNA mono-ubiquitination and activation of translesion DNA polymerases by DNA polymerase α. J Biochem 2009; 146:13-21; PMID:19279190; http://dx.doi.org/10.1093/jb/mvp043
15. Thu YM, Bielinsky AK. Enigmatic roles of Mcm10 in DNA replication. Trends Biochem Sci 2013; 38:184-94; PMID:23332289; http://dx.doi.org/10.1016/j.tibs. 2012.12.003
16. Wohlschlegel JA, Dhar SK, Prokhorova TA, Dutta A, Walter JC. Xenopus Mcm10 binds to origins of DNA replication after Mcm2-7 and stimulates origin binding of Cdc45. Mol Cell 2002; 9:233-40; PMID:11864598; http://dx.doi.org/10. 1016/S1097-2765(02)00456-2
17. Heller RC, Kang S, Lam WM, Chen S, Chan CS, Bell SP. Eukaryotic origin-dependent DNA replication in vitro reveals sequential action of DDK and S-CDK kinases. Cell 2011; 146:80-91; PMID:21729781; http://dx.doi.org/10.1016/j. cell.2011.06.012
18. Watase G, Takisawa H, Kanemaki MT. Mcm10 plays a role in functioning of the eukaryotic replicative DNA helicase, Cdc45-Mcm-GINS. Curr Biol 2012; 22:343-9; PMID:22285032; http://dx.doi.org/10.1016/j.cub.2012.01.023
19. Kanke M, Kodama Y, Takahashi TS, Nakagawa T, Masukata H. Mcm10 plays an essential role in origin DNA unwinding after loading of the CMG components. EMBO J 2012; 31:2182-94; PMID:22433840; http://dx.doi.org/10.1038/emboj.2012.68
20. van Deursen F, Sengupta S, De Piccoli G, Sanchez-Diaz A, Labib K. Mcm10 associates with the loaded DNA helicase at replication origins and defines a novel step in its activation. EMBO J 2012; 31:2195-206; PMID:22433841; http://dx.doi.org/10.1038/emboj.2012.69
21. Haworth J, Alver RC, Anderson M, Bielinsky AK. Ubc4 and Not4 regulate steady-state levels of DNA polymerase-α to promote efficient and accurate DNA replication. Mol Biol Cell 2010; 21:3205-19; PMID:20660159; http://dx.doi.org/10.1091/mbc.E09-06-0452
22. Robertson PD, Chagot B, Chazin WJ, Eichman BF. Solution NMR structure of the C-terminal DNA binding domain of Mcm10 reveals a conserved MCM motif. J Biol Chem 2010; 285:22942-9; PMID:20489205; http://dx.doi.org/10.1074/jbc.M110. 131276
23. Lee C, Liachko I, Bouten R, Kelman Z, Tye BK. Alternative mechanisms for coordinating polymerase α and MCM helicase. Mol Cell Biol 2010; 30:423-35; PMID:19917723; http://dx.doi.org/10.1128/MCB.01240-09
24. Warren EM, Huang H, Fanning E, Chazin WJ, Eichman BF. Physical interactions between Mcm10, DNA, and DNA polymerase α. J Biol Chem 2009; 284:24662-72; PMID:19608746; http://dx.doi.org/10.1074/jbc.M109.020438
25. Zhu W, Ukomadu C, Jha S, Senga T, Dhar SK, Wohlschlegel JA, Nutt LK, Kornbluth S, Dutta A. Mcm10 and And-1/CTF4 recruit DNA polymerase α to chromatin for initiation of DNA replication. Genes Dev 2007; 21:2288-99; PMID:17761813; http://dx.doi.org/10.1101/gad.1585607
26. Chattopadhyay S, Bielinsky AK. Human Mcm10 regulates the catalytic subunit of DNA polymerase-α and prevents DNA damage during replication. Mol Biol Cell 2007; 18:4085-95; PMID:17699597; http://dx.doi.org/10.1091/mbc. E06-12-1148
27. Ricke RM, Bielinsky AK. A conserved Hsp10-like domain in Mcm10 is required to stabilize the catalytic subunit of DNA polymerase-α in budding yeast. J Biol Chem 2006; 281:18414-25; PMID:16675460; http://dx.doi.org/10. 1074/jbc.M513551200
28. Ricke RM, Bielinsky AK. Mcm10 regulates the stability and chromatin association of DNA polymerase-α. Mol Cell 2004; 16:173-85; PMID:15494305; http://dx.doi.org/10.1016/j.molcel.2004.09.017
29. Fien K, Cho Y-S, Lee J-K, Raychaudhuri S, Tappin I, Hurwitz J. Primer utilization by DNA polymerase α-primase is influenced by its interaction with Mcm10p. J Biol Chem 2004; 279:16144-53; PMID:14766746; http://dx.doi.org/ 10.1074/jbc.M400142200
30. Yang X, Gregan J, Lindner K, Young H, Kearsey SE. Nuclear distribution and chromatin association of DNA polymerase α-primase is affected by TEV protease cleavage of Cdc23 (Mcm10) in fission yeast. BMC Mol Biol 2005; 6:13; PMID:15941470; http://dx.doi.org/10.1186/1471-2199-6-13
31. Das-Bradoo S, Ricke RM, Bielinsky AK. Interaction between PCNA and diubiquitinated Mcm10 is essential for cell growth in budding yeast. Mol Cell Biol 2006; 26:4806-17; PMID:16782870; http://dx.doi.org/10.1128/MCB.02062-05
32. Pacek M, Tutter AV, Kubota Y, Takisawa H, Walter JC. Localization of MCM2-7, Cdc45, and GINS to the site of DNA unwinding during eukaryotic DNA replication. Mol Cell 2006; 21:581-7; PMID:16483939; http://dx.doi.org/10.1016/ j.molcel.2006.01.030
33. Raveendranathan M, Chattopadhyay S, Bolon Y-T, Haworth J, Clarke DJ, Bielinsky AK. Genome-wide replication profiles of S-phase checkpoint mutants reveal fragile sites in yeast. EMBO J 2006; 25:3627-39; PMID:16888628; http://dx.doi.org/10.1038/sj.emboj.7601251
34. Taylor M, Moore K, Murray J, Aves SJ, Price C. Mcm10 interacts with Rad4/Cut5(TopBP1) and its association with origins of DNA replication is dependent on Rad4/Cut5(TopBP1). DNA Repair (Amst) 2011; 10:1154-63; PMID:21945095; http://dx.doi.org/10.1016/j.dnarep.2011.09.001
35. Zou L, Stillman B. Assembly of a complex containing Cdc45p, replication protein A, and Mcm2p at replication origins controlled by S-phase cyclin-dependent kinases and Cdc7p-Dbf4p kinase. Mol Cell Biol 2000; 20:3086-96; PMID:10757793; http://dx.doi.org/10.1128/MCB.20.9.3086-3096.2000
36. Varrin AE, Prasad AA, Scholz R-P, Ramer MD, Duncker BP. A mutation in Dbf4 motif M impairs interactions with DNA replication factors and confers increased resistance to genotoxic agents. Mol Cell Biol 2005; 25:7494-504; PMID:16107698; http://dx.doi.org/10.1128/MCB.25.17.7494-7504.2005
37. Lucchini G, Mazza C, Scacheri E, Plevani P. Genetic mapping of the Saccharomyces cerevisiae DNA polymerase I gene and characterization of a pol1 temperature-sensitive mutant altered in DNA primase-polymerase complex stability. Mol Gen Genet 1988; 212:459-65; PMID:3047550; http://dx.doi.org/10. 1007/BF00330850
38. Lucchini G, Muzi Falconi M, Pizzagalli A, Aguilera A, Klein HL, Plevani P. Nucleotide sequence and characterization of temperature-sensitive pol1 mutants of Saccharomyces cerevisiae. Gene 1990; 90:99-104; PMID:2199334; http://dx.doi.org/10.1016/0378-1119(90)90444-V
39. Pizzagalli A, Valsasnini P, Plevani P, Lucchini G. DNA polymerase I gene of Saccharomyces cerevisiae : nucleotide sequence, mapping of a temperature-sensitive mutation, and protein homology with other DNA polymerases. Proc Natl Acad Sci U S A 1988; 85:3772-6; PMID:3287376; http://dx.doi.org/10. 1073/pnas.85.11.3772
40. Gutiérrez PJ, Wang TS-F. Genomic instability induced by mutations in Saccharomyces cerevisiae POL1. Genetics 2003; 165:65-81; PMID:14504218
41. Schweitzer JK, Livingston DM. Expansions of CAG repeat tracts are frequent in a yeast mutant defective in Okazaki fragment maturation. Hum Mol Genet 1998; 7:69-74; PMID:9384605; http://dx.doi.org/10.1093/hmg/7.1.69
42. Homesley L, Lei M, Kawasaki Y, Sawyer S, Christensen T, Tye BK. Mcm10 and the MCM2-7 complex interact to initiate DNA synthesis and to release replication factors from origins. Genes Dev 2000; 14:913-26; PMID:10783164
43. Sawyer SL, Cheng IH, Chai W, Tye BK. Mcm10 and Cdc45 cooperate in origin activation in Saccharomyces cerevisiae . J Mol Biol 2004; 340:195-202; PMID:15201046; http://dx.doi.org/10.1016/j.jmb.2004.04.066
44. Ulrich HD. SUMO protocols. Humana Press New York, NY, 2009. 324 p.
45. Das-Bradoo S, Nguyen HD, Wood JL, Ricke RM, Haworth JC, Bielinsky AK. Defects in DNA ligase I trigger PCNA ubiquitylation at Lys 107. Nat Cell Biol 2010; 12:74-9, 1-20; PMID:20010813; http://dx.doi.org/10.1038/ncb2007
46. Nguyen HD, Becker J, Thu YM, Costanzo M, Koch EN, Smith S, Myung K, Myers CL, Boone C, Bielinsky AK. Unligated Okazaki fragments induce pcna ubiquitination and a requirement for rad59-dependent replication fork progression. PLoS One 2013; 8:e66379; PMID:23824283; http://dx.doi.org/10.1371/ journal.pone.0066379
47. Das-Bradoo S, Nguyen HD, Bielinsky AK. Damage-specific modification of PCNA. Cell Cycle 2010; 9:3674-9; PMID:20930510; http://dx.doi.org/10.4161/cc.9. 18.13121
48. Stelter P, Ulrich HD. Control of spontaneous and damage-induced mutagenesis by SUMO and ubiquitin conjugation. Nature 2003; 425:188-91; PMID:12968183; http://dx.doi.org/10.1038/nature01965
49. Makarova AV, Stodola JL, Burgers PM. A four-subunit DNA polymerase ζ complex containing Pol δ accessory subunits is essential for PCNA-mediated mutagenesis. Nucleic Acids Res 2012; 40:11618-26; PMID:23066099; http://dx.doi.org/10.1093/nar/gks948
50. Baranovskiy AG, Lada AG, Siebler HM, Zhang Y, Pavlov YI, Tahirov TH. DNA polymerase δ and ζ switch by sharing accessory subunits of DNA polymerase δ. J Biol Chem 2012; 287:17281-7; PMID:22465957; http://dx.doi.org/10.1074/jbc.M112.351122
51. Lei M, Kawasaki Y, Young MR, Kihara M, Sugino A, Tye BK. Mcm2 is a target of regulation by Cdc7-Dbf4 during the initiation of DNA synthesis. Genes Dev 1997; 11:3365-74; PMID:9407029; http://dx.doi.org/10.1101/gad.11.24.3365
52. Sheu Y-J, Stillman B. Cdc7-Dbf4 phosphorylates MCM proteins via a docking site-mediated mechanism to promote S phase progression. Mol Cell 2006; 24:101-13; PMID:17018296; http://dx.doi.org/10.1016/j.molcel.2006.07.033
53. Sheu Y-J, Stillman B. The Dbf4-Cdc7 kinase promotes S phase by alleviating an inhibitory activity in Mcm4. Nature 2010; 463:113-7; PMID:20054399; http://dx.doi.org/10.1038/nature08647
54. Pessoa-Brandão L, Sclafani RA. CDC7/DBF4 functions in the translesion synthesis branch of the RAD6 epistasis group in Saccharomyces cerevisiae. Genetics 2004; 167:1597-610; PMID:15342501; http://dx.doi.org/10. 1534/genetics.103.021675
55. Harkins V, Gabrielse C, Haste L, Weinreich M. Budding yeast Dbf4 sequences required for Cdc7 kinase activation and identification of a functional relationship between the Dbf4 and Rev1 BRCT domains. Genetics 2009; 183:1269-82; PMID:19822727; http://dx.doi.org/10.1534/genetics.109.110155
56. Osborn AJ, Elledge SJ. Mrc1 is a replication fork component whose phosphorylation in response to DNA replication stress activates Rad53. Genes Dev 2003; 17:1755-67; PMID:12865299; http://dx.doi.org/10.1101/gad.1098303
57. Downs JA, Lowndes NF, Jackson SP. A role for Saccharomyces cerevisiae histone H2A in DNA repair. Nature 2000; 408:1001-4; PMID:11140636; http://dx.doi.org/10.1038/35050000
58. Zhong Y, Nellimoottil T, Peace JM, Knott SR, Villwock SK, Yee JM, Jancuska JM, Rege S, Tecklenburg M, Sclafani RA, et al. The level of origin firing inversely affects the rate of replication fork progression. J Cell Biol 2013; 201:373-83; PMID:23629964; http://dx.doi.org/10.1083/jcb.201208060
59. Lazzaro F, Novarina D, Amara F, Watt DL, Stone JE, Costanzo V, Burgers PM, Kunkel TA, Plevani P, Muzi-Falconi M. RNase H and postreplication repair protect cells from ribonucleotides incorporated in DNA. Mol Cell 2012; 45:99-110; PMID:22244334; http://dx.doi.org/10.1016/j.molcel.2011.12.019
60. Branzei D, Seki M, Enomoto T. Rad18/Rad5/Mms2-mediated polyubiquitination of PCNA is implicated in replication completion during replication stress. Genes Cells 2004; 9:1031-42; PMID:15507115; http://dx.doi.org/10.1111/j.1365- 2443.2004.00787.x
61. Waters LS, Walker GC. The critical mutagenic translesion DNA polymerase Rev1 is highly expressed during G(2)/M phase rather than S phase. Proc Natl Acad Sci U S A 2006; 103:8971-6; PMID:16751278; http://dx.doi.org/10.1073/pnas. 0510167103
62. Hishida T, Kubota Y, Carr AM, Iwasaki H. RAD6-RAD18-RAD5-pathway- dependent tolerance to chronic low-dose ultraviolet light. Nature 2009; 457:612-5; PMID:19079240; http://dx.doi.org/10.1038/nature07580
63. Karras GI, Fumasoni M, Sienski G, Vanoli F, Branzei D, Jentsch S. Noncanonical role of the 9-1-1 clamp in the error-free DNA damage tolerance pathway. Mol Cell 2013; 49:536-46; PMID:23260657; http://dx.doi.org/10.1016/j. molcel.2012.11.016
64. Huang D, Piening BD, Paulovich AG. The preference for error-free or error-prone postreplication repair in Saccharomyces cerevisiae exposed to low-dose methyl methanesulfonate is cell cycle dependent. Mol Cell Biol 2013; 33:1515-27; PMID:23382077; http://dx.doi.org/10.1128/MCB.01392-12
65. Karras GI, Jentsch S. The RAD6 DNA damage tolerance pathway operates uncoupled from the replication fork and is functional beyond S phase. Cell 2010; 141:255-67; PMID:20403322; http://dx.doi.org/10.1016/j.cell.2010.02.028
66. Acharya N, Johnson RE, Prakash S, Prakash L. Complex formation with Rev1 enhances the proficiency of Saccharomyces cerevisiae DNA polymerase ζ for mismatch extension and for extension opposite from DNA lesions. Mol Cell Biol 2006; 26:9555-63; PMID:17030609; http://dx.doi.org/10.1128/MCB.01671-06
67. Haracska L, Unk I, Johnson RE, Johansson E, Burgers PM, Prakash S, Prakash L. Roles of yeast DNA polymerases δ and ζ and of Rev1 in the bypass of abasic sites. Genes Dev 2001; 15:945-54; PMID:11316789; http://dx.doi.org/10.1101/gad.882301
68. Albertella MR, Lau A, O'Connor MJ. The overexpression of specialized DNA polymerases in cancer. DNA Repair (Amst) 2005; 4:583-93; PMID:15811630; http://dx.doi.org/10.1016/j.dnarep.2005.01.005
69. Lee G-H, Nishimori H, Sasaki Y, Matsushita H, Kitagawa T, Tokino T. Analysis of lung tumorigenesis in chimeric mice indicates the pulmonary adenoma resistance 2 (Par2) locus to operate in the tumor-initiation stage in a cell-autonomous manner: detection of polymorphisms in the Polι gene as a candidate for Par2. Oncogene 2003; 22:2374-82; PMID:12700672; http://dx.doi.org/10.1038/sj.onc.1206387
70. Lee GH, Matsushita H. Genetic linkage between Pol ι deficiency and increased susceptibility to lung tumors in mice. Cancer Sci 2005; 96:256-9; PMID:15904465; http://dx.doi.org/10.1111/j.1349-7006.2005.00042.x
71. Yang J, Chen Z, Liu Y, Hickey RJ, Malkas LH. Altered DNA polymerase ι expression in breast cancer cells leads to a reduction in DNA replication fidelity and a higher rate of mutagenesis. Cancer Res 2004; 64:5597-607; PMID:15313897; http://dx.doi.org/10.1158/0008-5472.CAN-04-0603
72. Sakiyama T, Kohno T, Mimaki S, Ohta T, Yanagitani N, Sobue T, Kunitoh H, Saito R, Shimizu K, Hirama C, et al. Association of amino acid substitution polymorphisms in DNA repair genes TP53, POLI, REV1 and LIG4 with lung cancer risk. Int J Cancer 2005; 114:730-7; PMID:15609317; http://dx.doi.org/10.1002/ ijc.20790
73. O-Wang J, Kawamura K, Tada Y, Ohmori H, Kimura H, Sakiyama S, Tagawa M. DNA polymerase κ, implicated in spontaneous and DNA damage-induced mutagenesis, is overexpressed in lung cancer. Cancer Res 2001; 61:5366-9; PMID:11454676
74. Lemée F, Bergoglio V, Fernandez-Vidal A, Machado-Silva A, Pillaire M-J, Bieth A, Gentil C, Baker L, Martin A-L, Leduc C, et al. DNA polymerase θ upregulation is associated with poor survival in breast cancer, perturbs DNA replication, and promotes genetic instability. Proc Natl Acad Sci U S A 2010; 107:13390-5; PMID:20624954; http://dx.doi.org/10.1073/pnas.0910759107
75. Lorenz MC, Muir RS, Lim E, McElver J, Weber SC, Heitman J. Gene disruption with PCR products in Saccharomyces cerevisiae . Gene 1995; 158:113-7; PMID:7789793; http://dx.doi.org/10.1016/0378-1119(95)00144-U
76. Kokoska RJ, Stefanovic L, Tran HT, Resnick MA, Gordenin DA, Petes TD. Destabilization of yeast micro- and minisatellite DNA sequences by mutations affecting a nuclease involved in Okazaki fragment processing (rad27) and DNA polymerase δ (pol3-t). Mol Cell Biol 1998; 18:2779-88; PMID:9566897
77. Foster PL. Methods for determining spontaneous mutation rates. Methods Enzymol 2006; 409:195-213; PMID:16793403; http://dx.doi.org/10.1016/S0076- 6879(05)09012-9
78. Drake JW. A constant rate of spontaneous mutation in DNA-based microbes. Proc Natl Acad Sci U S A 1991; 88:7160-4; PMID:1831267; http://dx.doi.org/10. 1073/pnas.88.16.7160
79. Mann HB, Whitney DR. On a test of whether one of two random variables is stochastically larger than the other. Ann Math Stat 1947; 18:50-60; http://dx.doi.org/10.1214/aoms/1177730491