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The Baker's Yeast Diploid Genome Is Remarkably Stable in Vegetative Growth and Meiosis


Accurate estimates of mutation rates provide critical information to analyze genome evolution and organism fitness. We used whole-genome DNA sequencing, pulse-field gel electrophoresis, and comparative genome hybridization to determine mutation rates in diploid vegetative and meiotic mutation accumulation lines of Saccharomyces cerevisiae. The vegetative lines underwent only mitotic divisions while the meiotic lines underwent a meiotic cycle every ∼20 vegetative divisions. Similar base substitution rates were estimated for both lines. Given our experimental design, these measures indicated that the meiotic mutation rate is within the range of being equal to zero to being 55-fold higher than the vegetative rate. Mutations detected in vegetative lines were all heterozygous while those in meiotic lines were homozygous. A quantitative analysis of intra-tetrad mating events in the meiotic lines showed that inter-spore mating is primarily responsible for rapidly fixing mutations to homozygosity as well as for removing mutations. We did not observe 1–2 nt insertion/deletion (in-del) mutations in any of the sequenced lines and only one structural variant in a non-telomeric location was found. However, a large number of structural variations in subtelomeric sequences were seen in both vegetative and meiotic lines that did not affect viability. Our results indicate that the diploid yeast nuclear genome is remarkably stable during the vegetative and meiotic cell cycles and support the hypothesis that peripheral regions of chromosomes are more dynamic than gene-rich central sections where structural rearrangements could be deleterious. This work also provides an improved estimate for the mutational load carried by diploid organisms.


Vyšlo v časopise: The Baker's Yeast Diploid Genome Is Remarkably Stable in Vegetative Growth and Meiosis. PLoS Genet 6(9): e32767. doi:10.1371/journal.pgen.1001109
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pgen.1001109

Souhrn

Accurate estimates of mutation rates provide critical information to analyze genome evolution and organism fitness. We used whole-genome DNA sequencing, pulse-field gel electrophoresis, and comparative genome hybridization to determine mutation rates in diploid vegetative and meiotic mutation accumulation lines of Saccharomyces cerevisiae. The vegetative lines underwent only mitotic divisions while the meiotic lines underwent a meiotic cycle every ∼20 vegetative divisions. Similar base substitution rates were estimated for both lines. Given our experimental design, these measures indicated that the meiotic mutation rate is within the range of being equal to zero to being 55-fold higher than the vegetative rate. Mutations detected in vegetative lines were all heterozygous while those in meiotic lines were homozygous. A quantitative analysis of intra-tetrad mating events in the meiotic lines showed that inter-spore mating is primarily responsible for rapidly fixing mutations to homozygosity as well as for removing mutations. We did not observe 1–2 nt insertion/deletion (in-del) mutations in any of the sequenced lines and only one structural variant in a non-telomeric location was found. However, a large number of structural variations in subtelomeric sequences were seen in both vegetative and meiotic lines that did not affect viability. Our results indicate that the diploid yeast nuclear genome is remarkably stable during the vegetative and meiotic cell cycles and support the hypothesis that peripheral regions of chromosomes are more dynamic than gene-rich central sections where structural rearrangements could be deleterious. This work also provides an improved estimate for the mutational load carried by diploid organisms.


Zdroje

1. FriedbergEC

McDanielLD

SchultzRA

2004 The role of endogenous and exogenous DNA damage and mutagenesis. Curr Opin Genet Dev 14 5 10

2. McCullochSD

KunkelTA

2008 The fidelity of DNA synthesis by eukaryotic replicative and translesion synthesis polymerases. Cell Res 18 148 161

3. LynchM

KoskellaB

SchaackS

2006 Mutation pressure and the evolution of organelle genomic architecture. Science 311 1727 1730

4. NeiM

2007 The new mutation theory of phenotypic evolution. Proc Nat Acad Sci USA 104 12235 12242

5. RifkinSA

HouleD

KimJ

WhiteKP

2005 A mutation accumulation assay reveals a broad capacity for rapid evolution of gene expression. Nature 438 220 223

6. MullerHJ

1932 Some genetic aspects of sex. Am Nat 66 118 138

7. MullerHJ

1964 The relation of recombination to mutational advance. Mutat Res 106 2 9

8. LynchM

BürgerR

ButcherD

GabrielW

1993 The mutational meltdown in asexual populations. J Hered 84 339 344

9. KeightleyPD

OttoSP

2006 Interference among deleterious mutations favors sex and recombination in finite populations. Nature 443 89 92

10. MagniGE

von BorstelRC

1962 Different rates of spontaneous mutation during mitosis and meiosis in yeast. Genetics 47 1097 1108

11. StrathernJN

ShaferBK

McGillCB

1995 DNA synthesis errors associated with double-strand-break repair. Genetics 140 965 972

12. RattrayAJ

McGillCB

ShaferBK

StrathernJN

2001 Fidelity of mitotic double-strand-break repair in Saccharomyces cerevisiae: a role for SAE2/COM1. Genetics 158 109 122

13. FelsensteinJ

1974 The evolutionary advantage of recombination. Genetics 78 737 756

14. GoddardMR

GodfrayHC

BurtA

2005 Sex increases the efficacy of natural selection in experimental yeast populations. Nature 434 636 640

15. BaerCF

MiyamotoMM

DenverDR

2007 Mutation rate variation in multicellular eukaryotes: causes and consequences. Nat Rev Genet 8 619 631

16. NishantKT

SinghND

AlaniE

2009 Genomic mutation rates: what high-throughput methods can tell us. Bioessays 31 912 920

17. LynchM

SungW

MorrisK

CoffeyN

LandryCR

2008 A genome-wide view of the spectrum of spontaneous mutations in yeast. Proc Natl Acad Sci USA 105 9272 9277

18. HicksWM

KimM

HaberJE

2010 Increased mutagenesis and unique mutation signature associated with mitotic gene conversion. Science 329 82 85

19. McGillCB

HolbeckSL

StrathernJN

1998 The chromosome bias of misincorporations during double-strand break repair is not altered in mismatch repair-defective strains of Saccharomyces cerevisiae. Genetics 148 1525 1533

20. KeeneyS

GirouxCN

KlecknerN

1997 Meiosis-specific DNA double-strand breaks are catalyzed by Spo11, a member of a widely conserved protein family. Cell 88 375 384

21. CromieGA

SmithGR

2007 Branching out: meiotic recombination and its regulation. Trends Cell Biol 17 448 455

22. BegunDJ

AquadroCF

1992 Levels of naturally occurring DNA polymorphism correlate with recombination rates in D. melanogaster. Nature 356 519 520

23. Jensen-SeamanMI

FureyTS

PayseurBA

LuY

RoskinKM

2004 Comparative recombination rates in the rat, mouse, and human genomes. Genome Res 14 528 538

24. LercherMJ

HurstLD

2002 Human SNP variability and mutation rate are higher in regions of high recombination. Trends Genet 18 337 340

25. NachmanMW

2001 Single nucleotide polymorphisms and recombination rate in humans. Trends Genet 17 481 485

26. ManceraE

BourgonR

BrozziA

HuberW

SteinmetzLM

2008 High-resolution mapping of meiotic crossovers and noncrossovers in yeast. Nature 454 479 485

27. KeinanA

ReichD

2010 Human population differentiation is strongly correlated with local recombination rate. PLoS Genet 6 e1000886

28. NoorMA

2008 Mutagenesis from meiotic recombination is not a primary driver of sequence divergence between Saccharomyces species. Mol Biol Evol 25 2439 2444

29. TsaiIJ

BurtA

KoufopanouV

2010 Conservation of recombination hotspots in yeast. Proc Natl Acad Sci USA 107 7847 7852

30. KnopM

2006 Evolution of the hemiascomycete yeasts: on life styles and the importance of inbreeding. Bioessays 28 696 708

31. KellerPJ

KnopM

2009 Evolution of mutational robustness in the yeast genome: a link to essential genes and meiotic recombination hotspots. PLoS Genet 5 e1000533

32. WlochDM

SzafraniecK

BortsRH

KoronaR

2001 Direct estimate of the mutation rate and the distribution of fitness effects in the yeast Saccharomyces cerevisiae. Genetics 159 441 452

33. Eyre-WalkerA

KeightleyPD

2007 The distribution of fitness effects of new mutations. Nat Rev Genet 8 610 618

34. TsaiIJ

BensassonD

BurtA

KoufopanouV

2008 Population genomics of the wild yeast Saccharomyces paradoxus: Quantifying the life cycle. Proc Natl Acad Sci USA 105 4957 4962

35. KaneSM

RothR

1974 Carbohydrate metabolism during ascospore development in yeast. J Bacteriol 118 8 14

36. DrakeJW

1991 A constant rate of spontaneous mutation in DNA-based microbes. Proc. Natl Acad Sci USA 88 7160 7164

37. KeightleyPD

TrivediU

ThomsonM

OliverF

KumarS

2009 Analysis of the genome sequences of three Drosophila melanogaster spontaneous mutation accumulation lines. Genome Res 19 1195 1201

38. DenverDR

DolanPC

WilhelmLJ

SungW

Ignacio Lucas-LledóJI

2009 A genome-wide view of Caenorhabditis elegans base-substitution mutation processes. Proc Natl Acad Sci USA 106 16310 16314

39. KorbelJO

UrbanAE

AffourtitJP

GodwinB

GrubertF

2007 Paired-end mapping reveals extensive structural variation in the human genome. Science 318 420 426

40. XieC

TammiMT

2009 CNV-seq, a new method to detect copy number variation using high-throughput sequencing: BMC Bioinformatics 10 80

41. ChiangDY

GetzG

JaffeDB

O'KellyMJ

ZhaoX

2009 High-resolution mapping of copy-number alterations with massively parallel sequencing. Nature Methods 6 99 103

42. KimJM

VanguriS

BoekeJD

GabrielA

VoytasDF

1998 Transposable elements and genome organization: a comprehensive survey of retrotransposons revealed by the complete Saccharomyces cerevisiae genome sequence. Genome Res 8 464 478

43. PinkelD

SegravesR

SudarD

ClarkS

PooleI

1998 High resolution analysis of DNA copy number variation using comparative genomic hybridization to microarrays. Nat Genet 20 207 211

44. Solinas-ToldoS

LampelS

StilgenbauerS

NickolenkoJ

BennerA

1997 Matrix-based comparative genomic hybridization: biochips to screen for genomic imbalances. Genes Chromosomes Cancer 20 399 407

45. StrathernJN

HerskowitzI

1979 Asymmetry and directionality in production of new cell types during clonal growth: the switching pattern of homothallic yeast. Cell 17 371 381

46. RuderferDM

PrattSC

SeidelHS

KruglyakL

2006 Population genomic analysis of outcrossing and recombination in yeast. Nat Genet 38 1077 1081

47. ZeylC

2009 The role of sex in fungal evolution. Curr Opin Microbiol 12 592 598

48. BuhlerC

BordeV

LichtenM

2007 Mapping meiotic single-strand DNA reveals a new landscape of DNA double-strand breaks in Saccharomyces cerevisiae. PLoS Biol 5 e324

49. TerasawaM

OgawaH

TsukamotoY

ShinoharaM

ShirahigeK

2007 Meiotic recombination-related DNA synthesis and its implications for cross-over and non-cross-over recombinant formation. Proc Natl Acad Sci USA 104 5965 5970

50. GersteinAC

ChunHJ

GrantA

OttoSP

2006 Genomic convergence toward diploidy in Saccharomyces cerevisiae. PLoS Genet 2 e145

51. LeePS

PetesTD

2010 Mitotic gene conversion events induced in G1-synchronized yeast cells by gamma rays are similar to spontaneous conversion events. Proc Natl Acad Sci USA 107 7383 7388

52. TranHT

KeenJD

KrickerM

ResnickMA

GordeninDA

1997 Hypermutability of Homonucleotide Runs in Mismatch Repair and DNA Polymerase Proofreading Yeast Mutants. Mol Cell Biol 17 2859 2865

53. GraggH

HarfeBD

Jinks-RobertsonS

2002 Base composition of mononucleotide runs affects DNA polymerase slippage and removal of frameshift intermediates by mismatch repair in Saccharomyces cerevisiae. Mol Cell Biol 22 8756 8762

54. LouisEJ

1995 The chromosome ends of Saccharomyces cerevisiae. Yeast 11 1553 1573

55. MeffordHC

TraskBJ

2002 The complex structure and dynamic evolution of human subtelomeres. Nat Rev Genet 3 91 102

56. LinardopoulouEV

WilliamsEM

FanY

FriedmanC

YoungJM

2005 Human subtelomeres are hot spots of interchromosomal recombination and segmental duplication. Nature 437 94 100

57. PrydeFE

GorhamHC

LouisEJ

1997 Chromosome ends: all the same under their caps. Curr Opin Genet Dev 7 822 828

58. ArguesoJL

CarazzolleMF

MieczkowskiPA

DuarteFM

NettoOV

2009 Genome structure of a Saccharomyces cerevisiae strain widely used in bioethanol production. Genome Res 19 2258 2270

59. HoodME

AntonovicsJ

2004 Mating within the meiotic tetrad and the maintenance of genomic heterozygosity. Genetics 166 1751 1759

60. GreigD

LeuJY

2009 Natural history of budding yeast. Curr Biol 19 R886 R890

61. ColuccioA

NeimanAM

2004 Interspore bridges: a new feature of the Saccharomyces cerevisiae spore wall. Microbiology 150 3189 3196

62. JohnsonLJ

AntonovicsJ

HoodME

2005 The evolution of intratetrad mating rates. Evolution 59 2525 2532

63. MiltonJB

1997 Selection in natural populations Oxford Oxford University Press

64. RoseMD

WinstonF

HieterP

1990 Methods in yeast genetics NY Cold Spring Harbor Laboratory Press

65. WachA

BrachatA

PohlmannR

PhilippsenP

1994 New heterologous modules for classical or PCR-based gene disruptions in Saccharomyces cerevisiae. Yeast 10 1793 1808

66. GoldsteinAL

McCuskerJH

1999 Three new dominant drug resistance cassettes for gene disruption in Saccharomyces cerevisiae. Yeast 15 1541 1553

67. ArguesoJL

WanatJ

GemiciZ

AlaniE

2004 Competing crossover pathways act during meiosis in Saccharomyces cerevisiae. Genetics 168 1805 1816

68. RasmussenAK

ChatterjeeA

RasmussenLJ

SinghKK

2003 Mitochondria-mediated nuclear mutator phenotype in Saccharomyces cerevisiae. Nucl Acids Res 31 3909 3917

69. ZerbinoDR

BirneyE

2008 Velvet: algorithms for de novo short read assembly using de Bruijn graphs. Genome Res 18 821 829

70. SimpsonJT

WongK

JackmanSD

ScheinJE

JonesSJ

2009 ABySS: a parallel assembler for short read sequence data. Genome Res 19 1117 1123

71. SommerDD

DelcherAL

SalzbergSL

PopM

2007 Minimus: a fast, lightweight genome assembler. BMC Bioinformatics 8 64

72. LitiG

CarterDM

MosesAM

WarringerJ

PartsL

2009 Population genomics of domestic and wild yeasts. Nature 458 337 341

73. AltschulSF

MaddenTL

SchäfferAA

ZhangJ

ZhangZ

1997 Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucl Acids Res 25 3389 3402

74. LiH

RuanJ

DurbinR

2008 Mapping short DNA sequencing reads and calling variants using mapping quality scores. Genome Res 18 1851 1858

75. BolstadBM

IrizarryRA

AstrandM

SpeedTP

2003 A comparison of normalization methods for high density oligonucleotide array data based on variance and bias. Bioinformatics 19 185 193

76. ClevelandWS

1979 Robust locally weighted regression and smoothing scatter plots. J Am Stat Assoc 74 829 836

77. ArguesoJL

WestmorelandJ

MieczkowskiPA

GawelM

PetesTD

2008 Double-strand breaks associated with repetitive DNA can reshape the genome. Proc Natl Acad Sci USA 105 11845 11850

78. LemoineFJ

DegtyarevaNP

LobachevK

PetesTD

2005 Chromosomal translocations in yeast induced by low levels of DNA polymerase a model for chromosome fragile sites. Cell 120 587 598

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