Oxidative Stress Is Not a Major Contributor to Somatic Mitochondrial DNA Mutations
The accumulation of somatic mitochondrial DNA (mtDNA) mutations is implicated in aging and common diseases of the elderly, including cancer and neurodegenerative disease. However, the mechanisms that influence the frequency of somatic mtDNA mutations are poorly understood. To develop a simple invertebrate model system to address this matter, we used the Random Mutation Capture (RMC) assay to characterize the age-dependent frequency and distribution of mtDNA mutations in the fruit fly Drosophila melanogaster. Because oxidative stress is a major suspect in the age-dependent accumulation of somatic mtDNA mutations, we also used the RMC assay to explore the influence of oxidative stress on the somatic mtDNA mutation frequency. We found that many of the features associated with mtDNA mutations in vertebrates are conserved in Drosophila, including a comparable somatic mtDNA mutation frequency (∼10−5), an increased frequency of mtDNA mutations with age, and a prevalence of transition mutations. Only a small fraction of the mtDNA mutations detected in young or old animals were G∶C to T∶A transversions, a signature of oxidative damage, and loss-of-function mutations in the mitochondrial superoxide dismutase, Sod2, had no detectable influence on the somatic mtDNA mutation frequency. Moreover, a loss-of-function mutation in Ogg1, which encodes a DNA repair enzyme that removes oxidatively damaged deoxyguanosine residues (8-hydroxy-2′-deoxyguanosine), did not significantly influence the somatic mtDNA mutation frequency of Sod2 mutants. Together, these findings indicate that oxidative stress is not a major cause of somatic mtDNA mutations. Our data instead suggests that somatic mtDNA mutations arise primarily from errors that occur during mtDNA replication. Further studies using Drosophila should aid in the identification of factors that influence the frequency of somatic mtDNA mutations.
Vyšlo v časopise:
Oxidative Stress Is Not a Major Contributor to Somatic Mitochondrial DNA Mutations. PLoS Genet 10(2): e32767. doi:10.1371/journal.pgen.1003974
Kategorie:
Research Article
prolekare.web.journal.doi_sk:
https://doi.org/10.1371/journal.pgen.1003974
Souhrn
The accumulation of somatic mitochondrial DNA (mtDNA) mutations is implicated in aging and common diseases of the elderly, including cancer and neurodegenerative disease. However, the mechanisms that influence the frequency of somatic mtDNA mutations are poorly understood. To develop a simple invertebrate model system to address this matter, we used the Random Mutation Capture (RMC) assay to characterize the age-dependent frequency and distribution of mtDNA mutations in the fruit fly Drosophila melanogaster. Because oxidative stress is a major suspect in the age-dependent accumulation of somatic mtDNA mutations, we also used the RMC assay to explore the influence of oxidative stress on the somatic mtDNA mutation frequency. We found that many of the features associated with mtDNA mutations in vertebrates are conserved in Drosophila, including a comparable somatic mtDNA mutation frequency (∼10−5), an increased frequency of mtDNA mutations with age, and a prevalence of transition mutations. Only a small fraction of the mtDNA mutations detected in young or old animals were G∶C to T∶A transversions, a signature of oxidative damage, and loss-of-function mutations in the mitochondrial superoxide dismutase, Sod2, had no detectable influence on the somatic mtDNA mutation frequency. Moreover, a loss-of-function mutation in Ogg1, which encodes a DNA repair enzyme that removes oxidatively damaged deoxyguanosine residues (8-hydroxy-2′-deoxyguanosine), did not significantly influence the somatic mtDNA mutation frequency of Sod2 mutants. Together, these findings indicate that oxidative stress is not a major cause of somatic mtDNA mutations. Our data instead suggests that somatic mtDNA mutations arise primarily from errors that occur during mtDNA replication. Further studies using Drosophila should aid in the identification of factors that influence the frequency of somatic mtDNA mutations.
Zdroje
1. McBrideHM, NeuspielM, WasiakS (2006) Mitochondria: more than just a powerhouse. Current Biology 16: R551–560.
2. WallaceDC (2012) Mitochondria and cancer. Nature reviews Cancer 12: 685–698.
3. WangC, YouleRJ (2009) The role of mitochondria in apoptosis. Annual Review of Genetics 43: 95–118.
4. BooreJL (1999) Animal mitochondrial genomes. Nucleic Acids Res 27: 1767–1780.
5. DiMauroS, HiranoM (2005) Mitochondrial encephalomyopathies: an update. Neuromuscular disorders : NMD 15: 276–286.
6. ParkCB, LarssonNG (2011) Mitochondrial DNA mutations in disease and aging. The Journal of cell biology 193: 809–818.
7. BenderA, KrishnanKJ, MorrisCM, TaylorGA, ReeveAK, et al. (2006) High levels of mitochondrial DNA deletions in substantia nigra neurons in aging and Parkinson disease. Nature Genetics 38: 515–517.
8. KraytsbergY, KudryavtsevaE, McKeeAC, GeulaC, KowallNW, et al. (2006) Mitochondrial DNA deletions are abundant and cause functional impairment in aged human substantia nigra neurons. Nature Genetics 38: 518–520.
9. de GreyAD (2009) How is mutant mitochondrial DNA clonally amplified? Much new evidence, still no answers. Rejuvenation Res 12: 217–219.
10. LarssonNG (2010) Somatic mitochondrial DNA mutations in mammalian aging. Annu Rev Biochem 79: 683–706.
11. HarmanD (1972) The biologic clock: the mitochondria? Journal of the American Geriatrics Society 20: 145–147.
12. BrierleyEJ, JohnsonMA, LightowlersRN, JamesOF, TurnbullDM (1998) Role of mitochondrial DNA mutations in human aging: implications for the central nervous system and muscle. Ann Neurol 43: 217–223.
13. GuoM (2010) What have we learned from Drosophila models of Parkinson's disease? Prog Brain Res 184: 3–16.
14. KohH, ChungJ (2012) PINK1 as a molecular checkpoint in the maintenance of mitochondrial function and integrity. Mol Cells 34: 7–13.
15. CallejaM, PenaP, UgaldeC, FerreiroC, MarcoR, et al. (1993) Mitochondrial DNA remains intact during Drosophila aging, but the levels of mitochondrial transcripts are significantly reduced. The Journal of Biological Chemistry 268: 18891–18897.
16. SchwarzeSR, WeindruchR, AikenJM (1998) Decreased mitochondrial RNA levels without accumulation of mitochondrial DNA deletions in aging Drosophila melanogaster. Mutat Res 382: 99–107.
17. YuiR, MatsuuraET (2006) Detection of deletions flanked by short direct repeats in mitochondrial DNA of aging Drosophila. Mutat Res 594: 155–161.
18. BielasJH, LoebLA (2005) Quantification of random genomic mutations. Nat Methods 2: 285–290.
19. VermulstM, BielasJH, LoebLA (2008) Quantification of random mutations in the mitochondrial genome. Methods 46: 263–268.
20. Martinez-AzorinF, CallejaM, Hernandez-SierraR, FarrCL, KaguniLS, et al. (2008) Over-expression of the catalytic core of mitochondrial DNA (mtDNA) polymerase in the nervous system of Drosophila melanogaster reduces median life span by inducing mtDNA depletion. J Neurochem 105: 165–176.
21. JoersP, JacobsHT (2013) Analysis of replication intermediates indicates that Drosophila melanogaster mitochondrial DNA replicates by a strand-coupled theta mechanism. PLoS One 8: e53249.
22. GrossNJ, GetzGS, RabinowitzM (1969) Apparent turnover of mitochondrial deoxyribonucleic acid and mitochondrial phospholipids in the tissues of the rat. J Biol Chem 244: 1552–1562.
23. JonesS, ChenWD, ParmigianiG, DiehlF, BeerenwinkelN, et al. (2008) Comparative lesion sequencing provides insights into tumor evolution. Proceedings of the National Academy of Sciences of the United States of America 105: 4283–4288.
24. ShendureJ, JiH (2008) Next-generation DNA sequencing. Nature Biotechnology 26: 1135–1145.
25. VermulstM, BielasJH, KujothGC, LadigesWC, RabinovitchPS, et al. (2007) Mitochondrial point mutations do not limit the natural lifespan of mice. Nat Genet 39: 540–543.
26. KhrapkoK, VijgJ (2009) Mitochondrial DNA mutations and aging: devils in the details? Trends Genet 25: 91–98.
27. GreavesLC, ReeveAK, TaylorRW, TurnbullDM (2012) Mitochondrial DNA and disease. J Pathol 226: 274–286.
28. MurphyMP (2009) How mitochondria produce reactive oxygen species. Biochem J 417: 1–13.
29. De BontR, van LarebekeN (2004) Endogenous DNA damage in humans: a review of quantitative data. Mutagenesis 19: 169–185.
30. HarmanD (1956) Aging: a theory based on free radical and radiation chemistry. Journal of gerontology 11: 298–300.
31. WallaceDC (2001) A mitochondrial paradigm for degenerative diseases and ageing. Novartis Found Symp 235: 247–263 discussion 263-246.
32. MishraPC, SinghAK, SuhaiS (2005) Interaction of singlet oxygen and superoxide radical anion with guanine and formation of its mutagenic modification 8-oxoguanine. International Journal of Quantum Chemistry 102: 282–301.
33. FredericoLA, KunkelTA, ShawBR (1990) A sensitive genetic assay for the detection of cytosine deamination: determination of rate constants and the activation energy. Biochemistry 29: 2532–2537.
34. GraziewiczMA, BienstockRJ, CopelandWC (2007) The DNA polymerase gamma Y955C disease variant associated with PEO and parkinsonism mediates the incorporation and translesion synthesis opposite 7,8-dihydro-8-oxo-2′-deoxyguanosine. Hum Mol Genet 16: 2729–2739.
35. DuttaroyA, PaulA, KunduM, BeltonA (2003) A Sod2 null mutation confers severely reduced adult life span in Drosophila. Genetics 165: 2295–2299.
36. PaulA, BeltonA, NagS, MartinI, GrotewielMS, et al. (2007) Reduced mitochondrial SOD displays mortality characteristics reminiscent of natural aging. Mech Ageing Dev 128: 706–716.
37. ChintapalliVR, WangJ, DowJA (2007) Using FlyAtlas to identify better Drosophila melanogaster models of human disease. Nat Genet 39: 715–720.
38. de Souza-PintoNC, EideL, HogueBA, ThyboT, StevnsnerT, et al. (2001) Repair of 8-oxodeoxyguanosine lesions in mitochondrial dna depends on the oxoguanine dna glycosylase (OGG1) gene and 8-oxoguanine accumulates in the mitochondrial dna of OGG1-defective mice. Cancer Res 61: 5378–5381.
39. DherinC, DizdarogluM, DoerflingerH, BoiteuxS, RadicellaJP (2000) Repair of oxidative DNA damage in Drosophila melanogaster: identification and characterization of dOgg1, a second DNA glycosylase activity for 8-hydroxyguanine and formamidopyrimidines. Nucleic acids research 28: 4583–4592.
40. McQuiltonP, St PierreSE, ThurmondJ, FlyBaseC (2012) FlyBase 101–the basics of navigating FlyBase. Nucleic Acids Res 40: D706–714.
41. AmeurA, StewartJB, FreyerC, HagstromE, IngmanM, et al. (2011) Ultra-deep sequencing of mouse mitochondrial DNA: mutational patterns and their origins. PLoS Genet 7: e1002028.
42. KennedySR, SalkJJ, SchmittMW, LoebLA (2013) Ultra-sensitive sequencing reveals an age-related increase in somatic mitochondrial mutations that are inconsistent with oxidative damage. PLoS Genet 9: e1003794.
43. WeiYH (1998) Oxidative stress and mitochondrial DNA mutations in human aging. Proc Soc Exp Biol Med 217: 53–63.
44. Ruiz-PesiniE, LottMT, ProcaccioV, PooleJC, BrandonMC, et al. (2007) An enhanced MITOMAP with a global mtDNA mutational phylogeny. Nucleic Acids Res 35: D823–828.
45. DaiDF, SantanaLF, VermulstM, TomazelaDM, EmondMJ, et al. (2009) Overexpression of catalase targeted to mitochondria attenuates murine cardiac aging. Circulation 119: 2789–2797.
46. BokovA, ChaudhuriA, RichardsonA (2004) The role of oxidative damage and stress in aging. Mech Ageing Dev 125: 811–826.
47. SpelbrinkJN, ToivonenJM, HakkaartGA, KurkelaJM, CooperHM, et al. (2000) In vivo functional analysis of the human mitochondrial DNA polymerase POLG expressed in cultured human cells. J Biol Chem 275: 24818–24828.
48. GoddardJM, WolstenholmeDR (1978) Origin and direction of replication in mitochondrial DNA molecules from Drosophila melanogaster. Proc Natl Acad Sci U S A 75: 3886–3890.
49. GoddardJM, WolstenholmeDR (1980) Origin and direction of replication in mitochondrial DNA molecules from the genus Drosophila. Nucleic Acids Res 8: 741–757.
50. SaitoS, TamuraK, AotsukaT (2005) Replication origin of mitochondrial DNA in insects. Genetics 171: 1695–1705.
51. WagnerJR, CadetJ (2010) Oxidation reactions of cytosine DNA components by hydroxyl radical and one-electron oxidants in aerated aqueous solutions. Acc Chem Res 43: 564–571.
52. KreutzerDA, EssigmannJM (1998) Oxidized, deaminated cytosines are a source of C→T transitions in vivo. Proc Natl Acad Sci U S A 95: 3578–3582.
53. ZhengW, KhrapkoK, CollerHA, ThillyWG, CopelandWC (2006) Origins of human mitochondrial point mutations as DNA polymerase gamma-mediated errors. Mutat Res 599: 11–20.
54. BeletskiiA, BhagwatAS (1998) Correlation between transcription and C to T mutations in the non-transcribed DNA strand. Biol Chem 379: 549–551.
55. FrancinoMP, ChaoL, RileyMA, OchmanH (1996) Asymmetries generated by transcription-coupled repair in enterobacterial genes. Science 272: 107–109.
56. FrancinoMP, OchmanH (2001) Deamination as the basis of strand-asymmetric evolution in transcribed Escherichia coli sequences. Mol Biol Evol 18: 1147–1150.
57. Haag-LiautardC, CoffeyN, HouleD, LynchM, CharlesworthB, et al. (2008) Direct estimation of the mitochondrial DNA mutation rate in Drosophila melanogaster. PLoS Biol 6: e204.
58. BernsteinC, BernsteinH, PayneCM, GarewalH (2002) DNA repair/pro-apoptotic dual-role proteins in five major DNA repair pathways: fail-safe protection against carcinogenesis. Mutat Res 511: 145–178.
59. de Souza-PintoNC, MasonPA, HashiguchiK, WeissmanL, TianJ, et al. (2009) Novel DNA mismatch-repair activity involving YB-1 in human mitochondria. DNA Repair (Amst) 8: 704–719.
60. GanetzkyB, WuCF (1982) Indirect Suppression Involving Behavioral Mutants with Altered Nerve Excitability in DROSOPHILA MELANOGASTER. Genetics 100: 597–614.
61. RozenS, SkaletskyH (2000) Primer3 on the WWW for general users and for biologist programmers. Methods in molecular biology 132: 365–386.
62. RogersHH, Griffiths-JonesS (2012) Mitochondrial pseudogenes in the nuclear genomes of Drosophila. PLoS One 7: e32593.
63. ZhangK, LiZ, JaiswalM, BayatV, XiongB, et al. (2013) The C8ORF38 homologue Sicily is a cytosolic chaperone for a mitochondrial complex I subunit. J Cell Biol 200: 807–820.
64. Cardozo-PelaezF, BrooksPJ, StedefordT, SongS, Sanchez-RamosJ (2000) DNA damage, repair, and antioxidant systems in brain regions: a correlative study. Free Radic Biol Med 28: 779–785.
65. SieversF, WilmA, DineenD, GibsonTJ, KarplusK, et al. (2011) Fast, scalable generation of high-quality protein multiple sequence alignments using Clustal Omega. Molecular Systems Biology 7: 539.
66. GoujonM, McWilliamH, LiW, ValentinF, SquizzatoS, et al. (2010) A new bioinformatics analysis tools framework at EMBL-EBI. Nucleic acids research 38: W695–699.
67. ClarosMG, VincensP (1996) Computational method to predict mitochondrially imported proteins and their targeting sequences. Eur J Biochem 241: 779–786.
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