#PAGE_PARAMS# #ADS_HEAD_SCRIPTS# #MICRODATA#

Rates of Gyrase Supercoiling and Transcription Elongation Control Supercoil Density in a Bacterial Chromosome


Gyrase catalyzes negative supercoiling of DNA in an ATP-dependent reaction that helps condense bacterial chromosomes into a compact interwound “nucleoid.” The supercoil density (σ) of prokaryotic DNA occurs in two forms. Diffusible supercoil density (σD) moves freely around the chromosome in 10 kb domains, and constrained supercoil density (σC) results from binding abundant proteins that bend, loop, or unwind DNA at many sites. Diffusible and constrained supercoils contribute roughly equally to the total in vivo negative supercoil density of WT cells, so σ = σCD. Unexpectedly, Escherichia coli chromosomes have a 15% higher level of σ compared to Salmonella enterica. To decipher critical mechanisms that can change diffusible supercoil density of chromosomes, we analyzed strains of Salmonella using a 9 kb “supercoil sensor” inserted at ten positions around the genome. The sensor contains a complete Lac operon flanked by directly repeated resolvase binding sites, and the sensor can monitor both supercoil density and transcription elongation rates in WT and mutant strains. RNA transcription caused (−) supercoiling to increase upstream and decrease downstream of highly expressed genes. Excess upstream supercoiling was relaxed by Topo I, and gyrase replenished downstream supercoil losses to maintain an equilibrium state. Strains with TS gyrase mutations growing at permissive temperature exhibited significant supercoil losses varying from 30% of WT levels to a total loss of σD at most chromosome locations. Supercoil losses were influenced by transcription because addition of rifampicin (Rif) caused supercoil density to rebound throughout the chromosome. Gyrase mutants that caused dramatic supercoil losses also reduced the transcription elongation rates throughout the genome. The observed link between RNA polymerase elongation speed and gyrase turnover suggests that bacteria with fast growth rates may generate higher supercoil densities than slow growing species.


Vyšlo v časopise: Rates of Gyrase Supercoiling and Transcription Elongation Control Supercoil Density in a Bacterial Chromosome. PLoS Genet 8(8): e32767. doi:10.1371/journal.pgen.1002845
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pgen.1002845

Souhrn

Gyrase catalyzes negative supercoiling of DNA in an ATP-dependent reaction that helps condense bacterial chromosomes into a compact interwound “nucleoid.” The supercoil density (σ) of prokaryotic DNA occurs in two forms. Diffusible supercoil density (σD) moves freely around the chromosome in 10 kb domains, and constrained supercoil density (σC) results from binding abundant proteins that bend, loop, or unwind DNA at many sites. Diffusible and constrained supercoils contribute roughly equally to the total in vivo negative supercoil density of WT cells, so σ = σCD. Unexpectedly, Escherichia coli chromosomes have a 15% higher level of σ compared to Salmonella enterica. To decipher critical mechanisms that can change diffusible supercoil density of chromosomes, we analyzed strains of Salmonella using a 9 kb “supercoil sensor” inserted at ten positions around the genome. The sensor contains a complete Lac operon flanked by directly repeated resolvase binding sites, and the sensor can monitor both supercoil density and transcription elongation rates in WT and mutant strains. RNA transcription caused (−) supercoiling to increase upstream and decrease downstream of highly expressed genes. Excess upstream supercoiling was relaxed by Topo I, and gyrase replenished downstream supercoil losses to maintain an equilibrium state. Strains with TS gyrase mutations growing at permissive temperature exhibited significant supercoil losses varying from 30% of WT levels to a total loss of σD at most chromosome locations. Supercoil losses were influenced by transcription because addition of rifampicin (Rif) caused supercoil density to rebound throughout the chromosome. Gyrase mutants that caused dramatic supercoil losses also reduced the transcription elongation rates throughout the genome. The observed link between RNA polymerase elongation speed and gyrase turnover suggests that bacteria with fast growth rates may generate higher supercoil densities than slow growing species.


Zdroje

1. HigginsNP, PeeblesCL, SuginoA, CozzarelliNR (1978) Purification of the subunits of Escherichia coli DNA gyrase and reconstitution of enzymatic activity. Proc Natl Acad Sci USA 75: 1773–1777.

2. RocaJ (1995) The mechanisms of DNA topoisomerases. Trends Biochem Sci 20: 156–160.

3. DiNardoS, VoelkelKA, SternglanzR, ReynoldsAE, WrightA (1982) Escherichia coli DNA topoisomerase I mutants have compensatory mutations in DNA gyrase genes. Cell 31: 43–51.

4. DroletM, BroccoliS, RalluF, HraikyC, FortinC, et al. (2003) The problem of hypernegative supercoiling and R-loop formation in transcription. Front Biosci 8: D210–D221.

5. DiGateRJ, MariansKJ (1988) Identification of a potent decatenating enzyme from Escherichia coli. J Biol Chem 263: 13366–13373.

6. ZechiedrichEL, KhodurskyAB, BachellierS, SchneiderR, ChenD, et al. (2000) Roles of topoisomerases in maintaining steady-state DNA supercoiling in Escherichia coli. J Biol Chem 275: 8103–8113.

7. EspeliO, MariansKJ (2004) Untangling intracellular DNA topology. Mol Microbiol 52: 925–931.

8. PangZ, ChenR, MannaD, HigginsNP (2005) A gyrase mutant with low activity disrupts supercoiling at the replication terminus. J Bacteriol 187: 7773–7783.

9. OguraT, NikiH, MoriH, MoritaM, HasegawaM, et al. (1990) Identification and characterization of gyrB mutants of Escherichic coli that are defective in partitioning of mini-F plasmids. J Bacteriol 172: 1562–1568.

10. ChampionK, HigginsNP (2007) Growth rate toxicity phenotypes and homeostatic supercoil control differentiate Escherichia coli from Salmonella enterica serovar Typhimurium. J Bacteriol 189: 5839–5849.

11. HigginsCF, DormanCJ, StirlingDA, WaddellL, BoothIR, et al. (1988) A physiological role for DNA supercoiling in the osmotic regulation of gene expression in S. typhimurium and E. coli. Cell 52: 569–584.

12. PeterBJ, ArsuagaJ, BreierAM, KhodurskyAB, BrownPO, et al. (2004) Genomic transcriptional response to loss of chromosomal supercoiling in Escherichia coli. Genome Biol 5: R87.

13. BookerBM, DengS, HigginsNP (2010) DNA topology of highly transcribed operons in Salmonella enterica serovar Typhimurium. Mol Microbiol 78: 1348–1364.

14. Cozzarelli NR, Wang JC (1990) DNA topology and its biological effects. Cold Spring Harbor, N.Y.: Cold Spring Harbor Press. 480 p.

15. SternglanzR, DiNardoS, VoelkelKA, NishimuraY, HirotaY, et al. (1981) Mutations in the gene coding for Escherichia coli DNA topoisomerase I affect transcription and transposition. Proc Natl Acad Sci USA 78: 2747–2751.

16. CrozatE, PhilippeN, LenskiRE, GeiselmannJ, SchneiderD (2005) Long-Term Experimental Evolution in Escherichia coli. XII. DNA Topology as a Key Target of Selection. Genetics 169: 523–532.

17. CrozatE, WinkworthC, GaffeJ, HallinPF, RileyMA, et al. (2010) Parallel genetic and phenotypic evolution of DNA superhelicity in experimental populations of Escherichia coli. Mol Biol Evol 27: 2113–2128.

18. SteckTR, PrussGJ, ManesSH, BurgL, DrlicaK (1984) DNA supercoiling in gyrase mutants. J Bacteriol 158: 397–403.

19. Higgins NP, Vologodskii A (2004) Topological behavior of plasmid DNA. In: Funnell BE, Phillips GJ, editors. Plasmid Biology. Washington D.C.: ASM Press. pp. 181–201.

20. HigginsNP, YangX, FuQ, RothJR (1996) Surveying a supercoil domain by using the γδ resolution system in Salmonella typhimurium. J Bacteriol 178: 2825–2835.

21. SteinR, DengS, HigginsNP (2005) Measuring chromosome dynamics on different timescales using resolvases with varying half-lives. Mol Microbiol 56: 1049–1061.

22. BenjaminKR, AbolaAP, KanaarR, CozzarelliNR (1996) Contributions of supercoiling to Tn3 resolvase and phage Mu Gin site-specific recombination. J Mol Biol 256: 50–65.

23. StarkWM, SherrattDJ, BoocockMR (1989) Site-specific recombination by Tn3 resolvase: topological changes in the forward and reverse reactions. Cell 58: 779–790.

24. GrindleyND, WhitesonKL, RicePA (2006) Mechanisms of site-specific recombination. Annu Rev Biochem 75: 567–605.

25. OramM, MarkoJF, HalfordSE (1997) Communications between distant sites on supercoiled DNA from non-exponential kinetics for DNA synapsis by resolvase. J Mol Biol 270: 396–412.

26. StaczekP, HigginsNP (1998) DNA gyrase and Topoisomerase IV modulate chromosome domain size in vivo. Mol Micro 29: 1435–1448.

27. MoulinL, RahmouniAR, BoccardF (2005) Topological insulators inhibit diffusion of transcription-induced positive supercoils in the chromosome of Escherichia coli. Mol Microbiol 55: 601–610.

28. HardyC, CozzarelliNR (2005) A genetic selection for supercoiling mutants of Escherichia coli reveals proteins implicated in chromosome structure. Mol Microbio 57: 1636–1652.

29. PettijohnDE, PfenningerO (1980) Supercoils in prokaryotic DNA restrained in vivo. Proc Natl Acad Sci USA 77: 1331–1335.

30. GamperHB, HearstJE (1982) A topological model for transcription based on unwinding angle analysis of E. coli RNA polymerase binary, initiation and ternary complexes. Cell 29: 81–90.

31. DengS, SteinRA, HigginsNP (2004) Transcription-induced barriers to supercoil diffusion in the Salmonella typhimurium chromosome. Proc Natl Acad Sci USA 101: 3398–3403.

32. DengS, SteinRA, HigginsNP (2005) Organization of supercoil domains and their reorganization by transcription. Mol Microbio 57: 1511–1521.

33. LiuLF, WangJC (1987) Supercoiling of the DNA template during transcription. Proc Natl Acad Sci USA 84: 7024–7027.

34. PatoML, BennettPM, von MeyenburgK (1973) Messenger ribonucleic acid synthesis and degradation in Escherichia coli during inhibition of translation. J Bacteriol 116: 710–718.

35. KhodurskyAB, PeterBJ, SchmidMB, DeRisiJ, BotsteinD, et al. (2000) Analysis of topoisomerase function in bacterial replication fork movement: Use of DNA microarrays. Proc Natl Acad Sci USA 97: 9419–9424.

36. FassD, BogdenCE, BergerJM (1999) Quaternary changes in topoisomerase II may direct orthogonal movement of two DNA strands. Nat Struct Biol 6: 322–326.

37. QiY, PeiJ, GrishinNV (2002) C-terminal domain of gyrase A is predicted to have a beta-propeller structure. Proteins 47: 258–264.

38. AravindL, LeipeDD, KooninEV (1998) Toprim–a conserved catalytic domain in type IA and II topoisomerases, DnaG-type primases, OLD family nucleases and RecR proteins. Nucleic Acids Res 26: 4205–4213.

39. DuttaR, InouyeM (2000) GHKL, an emergent ATPase/kinase superfamily. Trends Biochem Sci 25: 24–28.

40. PostowL, HardyCD, ArsuagaJ, CozzarelliNR (2004) Topological domain structure of the Escherichia coli chromosome. Genes Dev 18: 1766–1779.

41. ValensM, PenaudS, RossignolM, CornetF, BoccardF (2004) Macrodomain organization of the Escherichia coli chromosome. EMBO J 23: 4330–4341.

42. NikiH, YamaichiY, HiragaS (2000) Dynamic organization of chromosomal DNA in Escherichia coli. Genes Dev 14: 212–223.

43. EsnaultE, ValensM, EspeliO, BoccardF (2007) Chromosome structuring limits genome plasticity in Escherichia coli. PLoS Genet 3: e226 doi:10.1371/journal.pgen.0030226.

44. EspeliO, MercierR, BoccardF (2008) DNA dynamics vary according to macrodomain topography in the E. coli chromosome. Mol Microbiol 68: 1418–1427.

45. Garcia-RussellN, HarmonTG, LeTQ, AmaladasNH, MathewsonRD, et al. (2004) Unequal access of chromosomal regions to each other in Salmonella : probing chromosome structure with phage l integrase-mediated long-range rearrangements. Mol Microbiol 52: 329–344.

46. LeeAK, DetweilerCS, FalkowS (2000) OmpR regulates the two-component system SsrA-SsrB in Salmonella pathogenicity island 2. J Bacteriol 182: 771–781.

47. MercierR, PetitMA, SchbathS, RobinS, El KarouiM, et al. (2008) The MatP/matS site-specific system organizes the terminus region of the E. coli chromosome into a macrodomain. Cell 135: 475–485.

48. AusselL, BarreF-X, AryoyM, StasiakA, StasiakAZ, et al. (2002) FtsK is a DNA motor protein that activates chromosome dimer resolution by switching the catalytic state of the XerC and XerD recombinases. Cell 108: 195–205.

49. HojgaardA, SzerlongH, taborC, KuempelP (1999) Norfloxacin-induced DNA cleavage occurs at the dif resolvase locus in Escherichia coli and is the result of interaction with topoisomerase IV. Mol Microbiol 33: 1027–1036.

50. BlakelyG, CollomsS, MayG, BurkeM, SherrattD (1991) Escherichia coli XerC recombinase is required for chromosomal segregation at cell division. The New Biologist 3: 789–798.

51. WangJC (1985) DNA topoisomerases. Annu Rev Biochem 54: 665–697.

52. HolmesVF, CozzarelliNR (2000) Closing the ring: Links between SMC proteins and chromosome partitioning, condensation, and supercoiling. Proc Natl Acad Sci USA 97: 1322–1324.

53. Brewer BJ (1990) Replication and the transcriptional organization of the Escherichia coli chromosome. In: Drlica K, Riley M, editors. The Bacterial Chromosome. Washington, D.C.: ASM Press. pp. 61–84.

54. BartlettMS, GaalT, RossW, GourseRL (1998) RNA polymerase mutants that destabilize RNA polymerase-promoter complexes alter NTP-sensing by rrn P1 promoters. J Mol Biol 279: 331–345.

55. VogelU, SorensenM, PedersenS, JensenKF, KilstrupM (1992) Decreasing transcription elongation rate in Escherichia coli exposed to amino acid starvation. Mol Microbiol 6: 2191–2200.

56. DameRT, KalmykowaOJ, GraingerDC (2011) Chromosomal macrodomains and associated proteins: implications for DNA organization and replication in gram negative bacteria. PLoS Genet 7: e1002123 doi:10.1371/journal.pgen.1002123.

57. DameRT, NoomMC, WuiteGJ (2006) Bacterial chromatin organization by H-NS protein unravelled using dual DNA manipulation. Nature 444: 387–390.

58. SkokoD, YooD, BaiH, SchnurrB, YanJ, et al. (2006) Mechanism of Chromosome Compaction and Looping by the Escherichia coli Nucleoid Protein Fis. J Mol Biol

59. CozzarelliNR (1980) DNA gyrase and the supercoiling of DNA. Science 207: 953–960.

60. MenzelR, GellertM (1983) Regulation of the genes for E. coli DNA gyrase: homeostatic control of DNA supercoiling. Cell 34: 105–113.

61. Tse-DinhY-C (1985) Regulation of the Escherichia coli DNA topoisomerase I gene by DNA supercoiling. Nucleic Acids Res 13.

62. DormanCJ, BhriainNN, HigginsCF (1990) DNA supercoiling and environmental regulation of virulence gene expression in Shigella flexneri. Nature 344: 789–792.

63. JensenPR, van der WeijdenCC, JensenLB, WesterhoffHV, SnoepJL (1999) Extensive regulation compromises the extent to which DNA gyrase controls DNA supercoiling and growth rate of Escherichia coli. Eur J Biochem 226: 865–877.

64. SnoepJL, van der WeijdenCC, AndersenHW, WesterhoffHV, JensenPR (2002) DNA supercoiling in Escherichia coli is under tight and subtle homeostatic control, involving gene-expression and metabolic regulation of both topoisomerase I and DNA gyrase. Eur J Biochem 269: 1662–1669.

65. SobetzkoP, TraversA, MuskhelishviliG (2012) Gene order and chromosome dynamics coordinate spatiotemporal gene expression during the bacterial growth cycle. Proc Natl Acad Sci U S A 109: E42–50.

66. MillerWG, SimonsRW (1993) Chromosomal supercoiling in Escherichia coli. Mol Microbiol 10: 675–684.

67. PavittGD, HigginsCF (1993) Chromosomal domains of supercoiling in Salmonella typhimurium. Mol Microbiol 10: 685–696.

68. HayamaR, MariansKJ (2010) Physical and functional interaction between the condensin MukB and the decatenase topoisomerase IV in Escherichia coli. Proc Natl Acad Sci U S A 107: 18826–18831.

69. WangX, Reyes-LamotheR, SherrattDJ (2008) Modulation of Escherichia coli sister chromosome cohesion by topoisomerase IV. Genes Dev 22: 2426–2433.

70. TehranchiAK, BlankschienMD, ZhangY, HallidayJA, SrivatsanA, et al. (2010) The transcription factor DksA prevents conflicts between DNA replication and transcription machinery. Cell 141: 595–605.

71. PaulBJ, BarkerMM, RossW, SchneiderDA, WebbC, et al. (2004) DksA: A critical component of the transcription initiation machinery that potentiates the regulation of rRNA promoters by ppGpp and the initiating NTP. Cell 118: 311–322.

72. Blanc-PotardAB, GariE, SpiritoF, Figueroa-BossiN, BossiL (1995) RNA polymerase (rpoB) mutants selected for increased resistance to gyrase inhibitors in Salmonella typhimurium. Mol Gen Genet 247: 680–692.

73. DormanCJ (1991) DNA supercoiling and environmental regulation of gene expression in pathogenic bacteria. Infect Immun 59: 745–749.

74. WuH-Y, ShyyS, WangJC, LiuLF (1988) Transcription generates positively and negatively supercoiled domains in the template. Cell 53: 433–440.

75. PrussG, DrlicaK (1986) Topoisomerase I mutants: The gene on pBR322 that encodes resistance to tetracycline affects plasmid DNA supercoiling. Proc Natl Acad Sci USA 83: 8952–8956.

76. SpiritoF, Figueroa-BossiN, BossiL (1994) The relative contributions of transcription and translation to plasmid DNA supercoiling in Salmonella typhimurium. Mol Microbiol 11: 111–122.

77. TretterEM, BergerJM (2012) Mechanisms for defining the supercoiling setpoint of DNA gyrase orthologs I. A non-conserved acidic C-terminal tail modulates E. coli gyrase activity. J Biol Chem

78. TretterEM, BergerJM (2012) Mechanisms For Defining Supercoiling Setpoint By DNA Gyrase Orthologs II. The shape of the GyrA CTD is not a sole determinant for controlling supercoiling efficiency. J Biol Chem

79. KleveczRR, BolenJ, ForrestG, MurrayDB (2004) A genomewide oscillation in transcription gates DNA replication and cell cycle. Proc Natl Acad Sci USA 101: 1200–1205.

80. ChenZ, OdstrcilEA, TuBP, McKnightSL (2007) Restriction of DNA replication to the reductive phase of the metabolic cycle protects genome integrity. Science 316: 1916–1919.

81. SilvermanSJ, PettiAA, SlavovN, ParsonsL, BriehofR, et al. (2010) Metabolic cycling in single yeast cells from unsynchronized steady-state populations limited on glucose or phosphate. Proc Natl Acad Sci U S A 107: 6946–6951.

82. StaraiVJ, CelicI, ColeRN, BoekeJD, Escalante-SemerenaJC (2002) Activation of acetyl-CoA synthetase by deacetylation of active lysine. Science 298: 2390–2392.

83. Bremer H, Dennis P (1996) Modulation of chemical composition and other parameters of the cell by growth rate. In: Neidhardt FC, editor. Escherichia coli and Salmonella typhimurium. Washington, DC: American Society for Microbiology Press. 1553.

84. EnglandJC, PerchukBS, LaubMT, GoberJW (2010) Global regulation of gene expression and cell differentiation in Caulobacter crescentus in response to nutrient availability. J Bacteriol 192: 819–833.

85. KosterDA, CrutA, ShumanS, BjornstiMA, DekkerNH (2010) Cellular strategies for regulating DNA supercoiling: a single-molecule perspective. Cell 142: 519–530.

86. MasonPB, StruhlK (2005) Distinction and relationship between elongation rate and processivity of RNA polymerase II in vivo. Mol Cell 17: 831–840.

87. GartenbergMR, WangJC (1992) Positive supercoiling of DNA greatly diminishes mRNA synthesis in yeast. Proc Natl Acad Sci U S A 89: 11461–11465.

88. YuD, EllisHE, LeeE-C, JenkinsNA, CopelandNG, et al. (2000) An efficient recombination system for chromosome engineering in Escherichia coli. Proc Natl Acad Sci USA 97: 5978–5983.

89. DattaS, CostantinoN, CourtDL (2006) A set of recombineering plasmids for gram-negative bacteria. Gene 379: 109–115.

90. JensenKF (1993) The Escherichia coli K-12 “wild types” W3110 and MG1655 have an rph frameshift mutation that leads to pyrimidine starvation due to low pyrE expression levels. J Bacteriol 175: 3401–3407.

91. Miller JH (1972) Experiments in Molecular Genetics; Miller JH, editor. Cold Spring Harbor, N.Y.: Cold Spring Harbor Laboratory.

92. SharanSK, ThomasonLC, KuznetsovSG, CourtDL (2009) Recombineering: a homologous recombination-based method of genetic engineering. Nat Protoc 4: 206–223.

Štítky
Genetika Reprodukčná medicína

Článok vyšiel v časopise

PLOS Genetics


2012 Číslo 8
Najčítanejšie tento týždeň
Najčítanejšie v tomto čísle
Kurzy

Zvýšte si kvalifikáciu online z pohodlia domova

Aktuální možnosti diagnostiky a léčby litiáz
nový kurz
Autori: MUDr. Tomáš Ürge, PhD.

Všetky kurzy
Prihlásenie
Zabudnuté heslo

Zadajte e-mailovú adresu, s ktorou ste vytvárali účet. Budú Vám na ňu zasielané informácie k nastaveniu nového hesla.

Prihlásenie

Nemáte účet?  Registrujte sa

#ADS_BOTTOM_SCRIPTS#