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The Genome Organization of Reflects Its Lifestyle


The generation of genome-scale data is becoming more routine, yet the subsequent analysis of omics data remains a significant challenge. Here, an approach that integrates multiple omics datasets with bioinformatics tools was developed that produces a detailed annotation of several microbial genomic features. This methodology was used to characterize the genome of Thermotoga maritima—a phylogenetically deep-branching, hyperthermophilic bacterium. Experimental data were generated for whole-genome resequencing, transcription start site (TSS) determination, transcriptome profiling, and proteome profiling. These datasets, analyzed in combination with bioinformatics tools, served as a basis for the improvement of gene annotation, the elucidation of transcription units (TUs), the identification of putative non-coding RNAs (ncRNAs), and the determination of promoters and ribosome binding sites. This revealed many distinctive properties of the T. maritima genome organization relative to other bacteria. This genome has a high number of genes per TU (3.3), a paucity of putative ncRNAs (12), and few TUs with multiple TSSs (3.7%). Quantitative analysis of promoters and ribosome binding sites showed increased sequence conservation relative to other bacteria. The 5′UTRs follow an atypical bimodal length distribution comprised of “Short” 5′UTRs (11–17 nt) and “Common” 5′UTRs (26–32 nt). Transcriptional regulation is limited by a lack of intergenic space for the majority of TUs. Lastly, a high fraction of annotated genes are expressed independent of growth state and a linear correlation of mRNA/protein is observed (Pearson r = 0.63, p<2.2×10−16 t-test). These distinctive properties are hypothesized to be a reflection of this organism's hyperthermophilic lifestyle and could yield novel insights into the evolutionary trajectory of microbial life on earth.


Vyšlo v časopise: The Genome Organization of Reflects Its Lifestyle. PLoS Genet 9(4): e32767. doi:10.1371/journal.pgen.1003485
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pgen.1003485

Souhrn

The generation of genome-scale data is becoming more routine, yet the subsequent analysis of omics data remains a significant challenge. Here, an approach that integrates multiple omics datasets with bioinformatics tools was developed that produces a detailed annotation of several microbial genomic features. This methodology was used to characterize the genome of Thermotoga maritima—a phylogenetically deep-branching, hyperthermophilic bacterium. Experimental data were generated for whole-genome resequencing, transcription start site (TSS) determination, transcriptome profiling, and proteome profiling. These datasets, analyzed in combination with bioinformatics tools, served as a basis for the improvement of gene annotation, the elucidation of transcription units (TUs), the identification of putative non-coding RNAs (ncRNAs), and the determination of promoters and ribosome binding sites. This revealed many distinctive properties of the T. maritima genome organization relative to other bacteria. This genome has a high number of genes per TU (3.3), a paucity of putative ncRNAs (12), and few TUs with multiple TSSs (3.7%). Quantitative analysis of promoters and ribosome binding sites showed increased sequence conservation relative to other bacteria. The 5′UTRs follow an atypical bimodal length distribution comprised of “Short” 5′UTRs (11–17 nt) and “Common” 5′UTRs (26–32 nt). Transcriptional regulation is limited by a lack of intergenic space for the majority of TUs. Lastly, a high fraction of annotated genes are expressed independent of growth state and a linear correlation of mRNA/protein is observed (Pearson r = 0.63, p<2.2×10−16 t-test). These distinctive properties are hypothesized to be a reflection of this organism's hyperthermophilic lifestyle and could yield novel insights into the evolutionary trajectory of microbial life on earth.


Zdroje

1. KitanoH (2002) Systems biology: a brief overview. Science 295: 1662–1664.

2. FeistAM, HerrgardMJ, ThieleI, ReedJL, PalssonBO (2009) Reconstruction of biochemical networks in microorganisms. Nat Rev Microbiol 7: 129–143.

3. ReedJL, FamiliI, ThieleI, PalssonBO (2006) Towards multidimensional genome annotation. Nat Rev Genet 7: 130–141.

4. OverbeekR, BartelsD, VonsteinV, MeyerF (2007) Annotation of bacterial and archaeal genomes: improving accuracy and consistency. Chem Rev 107: 3431–3447.

5. GuellM, van NoortV, YusE, ChenWH, Leigh-BellJ, et al. (2009) Transcriptome complexity in a genome-reduced bacterium. Science 326: 1268–1271.

6. KuhnerS, van NoortV, BettsMJ, Leo-MaciasA, BatisseC, et al. (2009) Proteome organization in a genome-reduced bacterium. Science 326: 1235–1240.

7. QiuY, ChoBK, ParkYS, LovleyD, PalssonBO, et al. (2010) Structural and operational complexity of the Geobacter sulfurreducens genome. Genome Res 20: 1304–1311.

8. SharmaCM, HoffmannS, DarfeuilleF, ReignierJ, FindeissS, et al. (2010) The primary transcriptome of the major human pathogen Helicobacter pylori. Nature 464: 250–255.

9. YoonSH, ReissDJ, BareJC, TenenbaumD, PanM, et al. (2011) Parallel evolution of transcriptome architecture during genome reorganization. Genome Res 21: 1892–1904.

10. BuescherJM, LiebermeisterW, JulesM, UhrM, MuntelJ, et al. (2012) Global network reorganization during dynamic adaptations of Bacillus subtilis metabolism. Science 335: 1099–1103.

11. NicolasP, MaderU, DervynE, RochatT, LeducA, et al. (2012) Condition-dependent transcriptome reveals high-level regulatory architecture in Bacillus subtilis. Science 335: 1103–1106.

12. SorekR, CossartP (2010) Prokaryotic transcriptomics: a new view on regulation, physiology and pathogenicity. Nat Rev Genet 11: 9–16.

13. PalssonB, ZenglerK (2010) The challenges of integrating multi-omic data sets. Nat Chem Biol 6: 787–789.

14. HuberR, LangworthyTA, KonigH, ThommM, WoeseCR, et al. (1986) Thermotoga maritima sp. nov. represents a new genus of unique extremely thermophilic eubacteria growing up to 90°C. Archives of Microbiology 144: 324–333.

15. DipippoJL, NesboCL, DahleH, DoolittleWF, BirklandNK, et al. (2009) Kosmotoga olearia gen. nov., sp. nov., a thermophilic, anaerobic heterotroph isolated from an oil production fluid. Int J Syst Evol Microbiol 59: 2991–3000.

16. NesboCL, DlutekM, ZhaxybayevaO, DoolittleWF (2006) Evidence for existence of “mesotogas,” members of the order Thermotogales adapted to low-temperature environments. Appl Environ Microbiol 72: 5061–5068.

17. NesboCL, KumaraswamyR, DlutekM, DoolittleWF, FoghtJ (2010) Searching for mesophilic Thermotogales bacteria: “mesotogas” in the wild. Appl Environ Microbiol 76: 4896–4900.

18. NesboCL, BradnanDM, AdebusuyiA, DlutekM, PetrusAK, et al. (2012) Mesotoga prima gen. nov., sp. nov., the first described mesophilic species of the Thermotogales. Extremophiles 16: 387–393.

19. ZhaxybayevaO, SwithersKS, FoghtJ, GreenAG, BruceD, et al. (2012) Genome Sequence of the Mesophilic Thermotogales Bacterium Mesotoga prima MesG1.Ag.4.2 Reveals the Largest Thermotogales Genome To Date. Genome Biol Evol 4: 700–708.

20. GiovannoniSJ, TrippHJ, GivanS, PodarM, VerginKL, et al. (2005) Genome streamlining in a cosmopolitan oceanic bacterium. Science 309: 1242–1245.

21. NelsonKE, ClaytonRA, GillSR, GwinnML, DodsonRJ, et al. (1999) Evidence for lateral gene transfer between archaea and bacteria from genome sequence of Thermotoga maritima. Nature 399: 323–329.

22. ConnersSB, MongodinEF, JohnsonMR, MonteroCI, NelsonKE, et al. (2006) Microbial biochemistry, physiology, and biotechnology of hyperthermophilic Thermotoga species. FEMS Microbiol Rev 30: 872–905.

23. MongodinEF, HanceIR, DeboyRT, GillSR, DaughertyS, et al. (2005) Gene transfer and genome plasticity in Thermotoga maritima, a model hyperthermophilic species. Journal of bacteriology 187: 4935–4944.

24. NesboCL, DlutekM, DoolittleWF (2006) Recombination in Thermotoga: implications for species concepts and biogeography. Genetics 172: 759–769.

25. ZhaxybayevaO, SwithersKS, LapierreP, FournierGP, BickhartDM, et al. (2009) On the chimeric nature, thermophilic origin, and phylogenetic placement of the Thermotogales. Proceedings of the National Academy of Sciences of the United States of America 106: 5865–5870.

26. MartinW, BarossJ, KelleyD, RussellMJ (2008) Hydrothermal vents and the origin of life. Nature reviews Microbiology 6: 805–814.

27. Achenbach-RichterL, GuptaR, StetterKO, WoeseCR (1987) Were the original eubacteria thermophiles? Systematic and applied microbiology 9: 34–39.

28. MunozR, YarzaP, LudwigW, EuzebyJ, AmannR, et al. (2011) Release LTPs104 of the All-Species Living Tree. Systematic and applied microbiology 34: 169–170.

29. FieldsPA (2001) Review: Protein function at thermal extremes: balancing stability and flexibility. Comp Biochem Physiol A Mol Integr Physiol 129: 417–431.

30. KumarS, NussinovR (2001) How do thermophilic proteins deal with heat? Cell Mol Life Sci 58: 1216–1233.

31. Gerday C, Glansdorff N, American Society for Microbiology. (2007) Physiology and biochemistry of extremophiles. Washington, D.C.: ASM Press. xvi, 429 p. p.

32. Robb FT (2008) Thermophiles : biology and technology at high temperatures. Boca Raton, FL: CRC Press. xiii, 353 p. p.

33. BoucherN, NollKM (2011) Ligands of thermophilic ABC transporters encoded in a newly sequenced genomic region of Thermotoga maritima MSB8 screened by differential scanning fluorimetry. Appl Environ Microbiol 77: 6395–6399.

34. AzizRK, BartelsD, BestAA, DeJonghM, DiszT, et al. (2008) The RAST Server: rapid annotations using subsystems technology. BMC genomics 9: 75.

35. ChoBK, ZenglerK, QiuY, ParkYS, KnightEM, et al. (2009) The transcription unit architecture of the Escherichia coli genome. Nat Biotechnol 27: 1043–1049.

36. VijayanV, JainIH, O'SheaEK (2011) A high resolution map of a cyanobacterial transcriptome. Genome Biol 12: R47.

37. KoideT, ReissDJ, BareJC, PangWL, FacciottiMT, et al. (2009) Prevalence of transcription promoters within archaeal operons and coding sequences. Mol Syst Biol 5: 285.

38. GruberAR, LorenzR, BernhartSH, NeubockR, HofackerIL (2008) The Vienna RNA websuite. Nucleic Acids Res 36: W70–74.

39. NawrockiEP, KolbeDL, EddySR (2009) Infernal 1.0: inference of RNA alignments. Bioinformatics 25: 1335–1337.

40. RossW, GosinkKK, SalomonJ, IgarashiK, ZouC, et al. (1993) A third recognition element in bacterial promoters: DNA binding by the alpha subunit of RNA polymerase. Science 262: 1407–1413.

41. BlatterEE, RossW, TangH, GourseRL, EbrightRH (1994) Domain organization of RNA polymerase alpha subunit: C-terminal 85 amino acids constitute a domain capable of dimerization and DNA binding. Cell 78: 889–896.

42. Schneider TD (1996) New Approaches In Mathematical Biology: Information Theory And Molecular Machines. In: Raulin JC-FaF, editor. Chemical Evolution: Physics of the Origin and Evolution of Life. Dordrecht, The Netherlands: Kluwer Academic Publishers. pp. 313–321.

43. SchneiderTD (1997) Information content of individual genetic sequences. J Theor Biol 189: 427–441.

44. D'HaeseleerP (2006) What are DNA sequence motifs? Nat Biotechnol 24: 423–425.

45. SchneiderTD (1991) Theory of molecular machines. II. Energy dissipation from molecular machines. J Theor Biol 148: 125–137.

46. ShultzabergerRK, RobertsLR, LyakhovIG, SidorovIA, StephenAG, et al. (2007) Correlation between binding rate constants and individual information of E. coli Fis binding sites. Nucleic Acids Res 35: 5275–5283.

47. RhodiusVA, MutalikVK (2010) Predicting strength and function for promoters of the Escherichia coli alternative sigma factor, sigmaE. Proc Natl Acad Sci U S A 107: 2854–2859.

48. ShultzabergerRK, ChenZ, LewisKA, SchneiderTD (2007) Anatomy of Escherichia coli sigma70 promoters. Nucleic Acids Res 35: 771–788.

49. KeselerIM, Collado-VidesJ, Santos-ZavaletaA, Peralta-GilM, Gama-CastroS, et al. (2011) EcoCyc: a comprehensive database of Escherichia coli biology. Nucleic Acids Res 39: D583–590.

50. KrogerC, DillonSC, CameronAD, PapenfortK, SivasankaranSK, et al. (2012) The transcriptional landscape and small RNAs of Salmonella enterica serovar Typhimurium. Proc Natl Acad Sci U S A 109: E1277–1286.

51. AlbrechtM, SharmaCM, DittrichMT, MullerT, ReinhardtR, et al. (2011) The transcriptional landscape of Chlamydia pneumoniae. Genome Biol 12: R98.

52. MitschkeJ, GeorgJ, ScholzI, SharmaCM, DienstD, et al. (2011) An experimentally anchored map of transcriptional start sites in the model cyanobacterium Synechocystis sp. PCC6803. Proc Natl Acad Sci U S A 108: 2124–2129.

53. SierroN, MakitaY, de HoonM, NakaiK (2008) DBTBS: a database of transcriptional regulation in Bacillus subtilis containing upstream intergenic conservation information. Nucleic Acids Res 36: D93–96.

54. ChenH, BjerknesM, KumarR, JayE (1994) Determination of the optimal aligned spacing between the Shine-Dalgarno sequence and the translation initiation codon of Escherichia coli mRNAs. Nucleic Acids Res 22: 4953–4957.

55. MolinaN, van NimwegenE (2008) Universal patterns of purifying selection at noncoding positions in bacteria. Genome Res 18: 148–160.

56. NelsonCE, HershBM, CarrollSB (2004) The regulatory content of intergenic DNA shapes genome architecture. Genome Biol 5: R25.

57. NovichkovPS, LaikovaON, NovichkovaES, GelfandMS, ArkinAP, et al. (2010) RegPrecise: a database of curated genomic inferences of transcriptional regulatory interactions in prokaryotes. Nucleic Acids Res 38: D111–118.

58. MaierT, SchmidtA, GuellM, KuhnerS, GavinAC, et al. (2011) Quantification of mRNA and protein and integration with protein turnover in a bacterium. Mol Syst Biol 7: 511.

59. NieL, WuG, ZhangW (2006) Correlation between mRNA and protein abundance in Desulfovibrio vulgaris: a multiple regression to identify sources of variations. Biochemical and biophysical research communications 339: 603–610.

60. TowseyM, HoganJM, MathewsS, TimmsP (2007) The in silico prediction of promoters in bacterial genomes. Genome Inform 19: 178–189.

61. RangannanV, BansalM (2011) PromBase: a web resource for various genomic features and predicted promoters in prokaryotic genomes. BMC Res Notes 4: 257.

62. GerlandU, MorozJD, HwaT (2002) Physical constraints and functional characteristics of transcription factor-DNA interaction. Proceedings of the National Academy of Sciences of the United States of America 99: 12015–12020.

63. GaltierN, LobryJR (1997) Relationships between genomic G+C content, RNA secondary structures, and optimal growth temperature in prokaryotes. J Mol Evol 44: 632–636.

64. FrockAD, GraySR, KellyRM (2012) Hyperthermophilic Thermotoga species differ with respect to specific carbohydrate transporters and glycoside hydrolases. Appl Environ Microbiol 78: 1978–1986.

65. DarfeuilleF, UnosonC, VogelJ, WagnerEG (2007) An antisense RNA inhibits translation by competing with standby ribosomes. Molecular cell 26: 381–392.

66. WatersLS, StorzG (2009) Regulatory RNAs in bacteria. Cell 136: 615–628.

67. RinkerKD, KellyRM (1996) Growth physiology of the hyperthermophilic Archaeon Thermococcus litoralis: development of a sulfur-free defined medium, characterization of an exopolysaccharide, and evidence of biofilm formation. Appl Environ Microbiol 62: 4478–4485.

68. PortnoyVA, HerrgardMJ, PalssonBO (2008) Aerobic fermentation of D-glucose by an evolved cytochrome oxidase-deficient Escherichia coli strain. Appl Environ Microbiol 74: 7561–7569.

69. PyszMA, WardDE, ShockleyKR, MonteroCI, ConnersSB, et al. (2004) Transcriptional analysis of dynamic heat-shock response by the hyperthermophilic bacterium Thermotoga maritima. Extremophiles 8: 209–217.

70. SchneebergerK, OssowskiS, LanzC, JuulT, PetersenAH, et al. (2009) SHOREmap: simultaneous mapping and mutation identification by deep sequencing. Nat Methods 6: 550–551.

71. ZerbinoDR, BirneyE (2008) Velvet: algorithms for de novo short read assembly using de Bruijn graphs. Genome Res 18: 821–829.

72. HalaszG, van BatenburgMF, PerusseJ, HuaS, LuXJ, et al. (2006) Detecting transcriptionally active regions using genomic tiling arrays. Genome Biol 7: R59.

73. LevinJZ, YassourM, AdiconisX, NusbaumC, ThompsonDA, et al. (2010) Comprehensive comparative analysis of strand-specific RNA sequencing methods. Nat Methods 7: 709–715.

74. LangmeadB, TrapnellC, PopM, SalzbergSL (2009) Ultrafast and memory-efficient alignment of short DNA sequences to the human genome. Genome Biol 10: R25.

75. TrapnellC, WilliamsBA, PerteaG, MortazaviA, KwanG, et al. (2010) Transcript assembly and quantification by RNA-Seq reveals unannotated transcripts and isoform switching during cell differentiation. Nat Biotechnol 28: 511–515.

76. Schrimpe-RutledgeAC, JonesMB, ChauhanS, PurvineSO, SanfordJA, et al. (2012) Comparative Omics-Driven Genome Annotation Refinement: Application across Yersiniae. PLoS ONE 7: e33903 doi:10.1371/journal.pone.0033903.

77. LermanJA, HydukeDR, LatifH, PortnoyVA, LewisNE, et al. (2012) In silico method for modelling metabolism and gene product expression at genome scale. Nat Commun 3: 929.

78. LiuX, BrutlagDL, LiuJS (2001) BioProspector: discovering conserved DNA motifs in upstream regulatory regions of co-expressed genes. Pacific Symposium on Biocomputing Pacific Symposium on Biocomputing 127–138.

79. BaileyTL, ElkanC (1994) Fitting a mixture model by expectation maximization to discover motifs in biopolymers. Proceedings/International Conference on Intelligent Systems for Molecular Biology ; ISMB International Conference on Intelligent Systems for Molecular Biology 2: 28–36.

80. CrooksGE, HonG, ChandoniaJM, BrennerSE (2004) WebLogo: a sequence logo generator. Genome Res 14: 1188–1190.

81. MarkhamNR, ZukerM (2008) UNAFold: software for nucleic acid folding and hybridization. Methods in molecular biology 453: 3–31.

82. TakemotoK, NacherJC, AkutsuT (2007) Correlation between structure and temperature in prokaryotic metabolic networks. BMC Bioinformatics 8: 303.

83. KingsfordCL, AyanbuleK, SalzbergSL (2007) Rapid, accurate, computational discovery of Rho-independent transcription terminators illuminates their relationship to DNA uptake. Genome Biol 8: R22.

84. GardnerPP, DaubJ, TateJ, MooreBL, OsuchIH, et al. (2011) Rfam: Wikipedia, clans and the “decimal” release. Nucleic Acids Res 39: D141–145.

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Genetika Reprodukčná medicína

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