#PAGE_PARAMS# #ADS_HEAD_SCRIPTS# #MICRODATA#

Chromosomal Organization and Segregation in


The study of chromosomal organization and segregation in a handful of bacteria has revealed surprising variety in the mechanisms mediating such fundamental processes. In this study, we further emphasized this diversity by revealing an original organization of the Pseudomonas aeruginosa chromosome. We analyzed the localization of 20 chromosomal markers and several components of the replication machinery in this important opportunistic γ-proteobacteria pathogen. This technique allowed us to show that the 6.3 Mb unique circular chromosome of P. aeruginosa is globally oriented from the old pole of the cell to the division plane/new pole along the oriC-dif axis. The replication machinery is positioned at mid-cell, and the chromosomal loci from oriC to dif are moved sequentially to mid-cell prior to replication. The two chromosomal copies are subsequently segregated at their final subcellular destination in the two halves of the cell. We identified two regions in which markers localize at similar positions, suggesting a bias in the distribution of chromosomal regions in the cell. The first region encompasses 1.4 Mb surrounding oriC, where loci are positioned around the 0.2/0.8 relative cell length upon segregation. The second region contains at least 800 kb surrounding dif, where loci show an extensive colocalization step following replication. We also showed that disrupting the ParABS system is very detrimental in P. aeruginosa. Possible mechanisms responsible for the coordinated chromosomal segregation process and for the presence of large distinctive regions are discussed.


Vyšlo v časopise: Chromosomal Organization and Segregation in. PLoS Genet 9(5): e32767. doi:10.1371/journal.pgen.1003492
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pgen.1003492

Souhrn

The study of chromosomal organization and segregation in a handful of bacteria has revealed surprising variety in the mechanisms mediating such fundamental processes. In this study, we further emphasized this diversity by revealing an original organization of the Pseudomonas aeruginosa chromosome. We analyzed the localization of 20 chromosomal markers and several components of the replication machinery in this important opportunistic γ-proteobacteria pathogen. This technique allowed us to show that the 6.3 Mb unique circular chromosome of P. aeruginosa is globally oriented from the old pole of the cell to the division plane/new pole along the oriC-dif axis. The replication machinery is positioned at mid-cell, and the chromosomal loci from oriC to dif are moved sequentially to mid-cell prior to replication. The two chromosomal copies are subsequently segregated at their final subcellular destination in the two halves of the cell. We identified two regions in which markers localize at similar positions, suggesting a bias in the distribution of chromosomal regions in the cell. The first region encompasses 1.4 Mb surrounding oriC, where loci are positioned around the 0.2/0.8 relative cell length upon segregation. The second region contains at least 800 kb surrounding dif, where loci show an extensive colocalization step following replication. We also showed that disrupting the ParABS system is very detrimental in P. aeruginosa. Possible mechanisms responsible for the coordinated chromosomal segregation process and for the presence of large distinctive regions are discussed.


Zdroje

1. SilbyMW, WinstanleyC, GodfreySA, LevySB, JacksonRW (2011) Pseudomonas genomes: diverse and adaptable. FEMS Microbiol Rev 35: 652–680.

2. GovanJR, DereticV (1996) Microbial pathogenesis in cystic fibrosis: mucoid Pseudomonas aeruginosa and Burkholderia cepacia. Microbiol Rev 60: 539–574.

3. ListerPD, WolterDJ, HansonND (2009) Antibacterial-resistant Pseudomonas aeruginosa: clinical impact and complex regulation of chromosomally encoded resistance mechanisms. Clin Microbiol Rev 22: 582–610.

4. StoverCK, PhamXQ, ErwinAL, MizoguchiSD, WarrenerP, et al. (2000) Complete genome sequence of Pseudomonas aeruginosa PAO1, an opportunistic pathogen. Nature 406: 959–964.

5. YeeTW, SmithDW (1990) Pseudomonas chromosomal replication origins: a bacterial class distinct from Escherichia coli-type origins. Proc Natl Acad Sci U S A 87: 1278–1282.

6. JiangY, YaoS, HelinskiD, ToukdarianA (2006) Functional analysis of two putative chromosomal replication origins from Pseudomonas aeruginosa. Plasmid 55: 194–200.

7. de VriesR (2010) DNA condensation in bacteria: Interplay between macromolecular crowding and nucleoid proteins. Biochimie 92: 1715–1721.

8. ThanbichlerM, ShapiroL (2006) Chromosome organization and segregation in bacteria. J Struct Biol 156: 292–303.

9. ToroE, ShapiroL (2010) Bacterial chromosome organization and segregation. Cold Spring Harb Perspect Biol 2: a000349.

10. PossozC, JunierI, EspeliO (2012) Bacterial chromosome segregation. Front Biosci 17: 1020–1034.

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

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

13. LemonKP, GrossmanAD (2001) The extrusion-capture model for chromosome partitioning in bacteria. Genes Dev 15: 2031–2041.

14. BatesD, KlecknerN (2005) Chromosome and replisome dynamics in E. coli: loss of sister cohesion triggers global chromosome movement and mediates chromosome segregation. Cell 121: 899–911.

15. Reyes-LamotheR, PossozC, DanilovaO, SherrattDJ (2008) Independent positioning and action of Escherichia coli replisomes in live cells. Cell 133: 90–102.

16. JunS, WrightA (2010) Entropy as the driver of chromosome segregation. Nat Rev Microbiol 8: 600–607.

17. FogelMA, WaldorMK (2006) A dynamic, mitotic-like mechanism for bacterial chromosome segregation. Genes Dev 20: 3269–3282.

18. ToroE, HongSH, McAdamsHH, ShapiroL (2008) Caulobacter requires a dedicated mechanism to initiate chromosome segregation. Proc Natl Acad Sci U S A 105: 15435–15440.

19. LeePS, LinDC, MoriyaS, GrossmanAD (2003) Effects of the chromosome partitioning protein Spo0J (ParB) on oriC positioning and replication initiation in Bacillus subtilis. J Bacteriol 185: 1326–1337.

20. MinnenA, AttaiechL, ThonM, GruberS, VeeningJW (2011) SMC is recruited to oriC by ParB and promotes chromosome segregation in Streptococcus pneumoniae. Mol Microbiol 81: 676–688.

21. SullivanNL, MarquisKA, RudnerDZ (2009) Recruitment of SMC by ParB-parS organizes the origin region and promotes efficient chromosome segregation. Cell 137: 697–707.

22. GruberS, ErringtonJ (2009) Recruitment of condensin to replication origin regions by ParB/SpoOJ promotes chromosome segregation in B. subtilis. Cell 137: 685–696.

23. PetrushenkoZM, SheW, RybenkovVV (2011) A new family of bacterial condensins. Mol Microbiol 81: 881–896.

24. LasockiK, BartosikAA, MierzejewskaJ, ThomasCM, Jagura-BurdzyG (2007) Deletion of the parA (soj) homologue in Pseudomonas aeruginosa causes ParB instability and affects growth rate, chromosome segregation, and motility. J Bacteriol 189: 5762–5772.

25. BartosikAA, MierzejewskaJ, ThomasCM, Jagura-BurdzyG (2009) ParB deficiency in Pseudomonas aeruginosa destabilizes the partner protein ParA and affects a variety of physiological parameters. Microbiology 155: 1080–1092.

26. LauIF, FilipeSR, SoballeB, OkstadOA, BarreFX, et al. (2003) Spatial and temporal organization of replicating Escherichia coli chromosomes. Mol Microbiol 49: 731–743.

27. NielsenHJ, OttesenJR, YoungrenB, AustinSJ, HansenFG (2006) The Escherichia coli chromosome is organized with the left and right chromosome arms in separate cell halves. Mol Microbiol 62: 331–338.

28. ViollierPH, ThanbichlerM, McGrathPT, WestL, MeewanM, et al. (2004) Rapid and sequential movement of individual chromosomal loci to specific subcellular locations during bacterial DNA replication. Proc Natl Acad Sci U S A 101: 9257–9262.

29. LemonKP, GrossmanAD (1998) Localization of bacterial DNA polymerase: evidence for a factory model of replication. Science 282: 1516–1519.

30. JensenRB, WangSC, ShapiroL (2001) A moving DNA replication factory in Caulobacter crescentus. EMBO J 20: 4952–4963.

31. LandgrafD, OkumusB, ChienP, BakerTA, PaulssonJ (2012) Segregation of molecules at cell division reveals native protein localization. Nat Methods 9: 480–482.

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

33. WangX, PossozC, SherrattDJ (2005) Dancing around the divisome: asymmetric chromosome segregation in Escherichia coli. Genes Dev 19: 2367–2377.

34. BartosikAA, LasockiK, MierzejewskaJ, ThomasCM, Jagura-BurdzyG (2004) ParB of Pseudomonas aeruginosa: interactions with its partner ParA and its target parS and specific effects on bacterial growth. J Bacteriol 186: 6983–6998.

35. Ben-YehudaS, RudnerDZ, LosickR (2003) RacA, a bacterial protein that anchors chromosomes to the cell poles. Science 299: 532–536.

36. BowmanGR, ComolliLR, ZhuJ, EckartM, KoenigM, et al. (2008) A polymeric protein anchors the chromosomal origin/ParB complex at a bacterial cell pole. Cell 134: 945–955.

37. EbersbachG, BriegelA, JensenGJ, Jacobs-WagnerC (2008) A self-associating protein critical for chromosome attachment, division, and polar organization in caulobacter. Cell 134: 956–968.

38. YamaichiY, BrucknerR, RinggaardS, MollA, CameronDE, et al. (2012) A multidomain hub anchors the chromosome segregation and chemotactic machinery to the bacterial pole. Genes Dev 26: 2348–2360.

39. LiY, YoungrenB, SergueevK, AustinS (2003) Segregation of the Escherichia coli chromosome terminus. Mol Microbiol 50: 825–834.

40. 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.

41. EspeliO, BorneR, DupaigneP, ThielA, GigantE, et al. (2012) A MatP-divisome interaction coordinates chromosome segregation with cell division in E. coli. EMBO J 31: 3198–3211.

42. JensenRB (2006) Coordination between chromosome replication, segregation, and cell division in Caulobacter crescentus. J Bacteriol 188: 2244–2253.

43. SrivastavaP, FeketeRA, ChattorajDK (2006) Segregation of the replication terminus of the two Vibrio cholerae chromosomes. J Bacteriol 188: 1060–1070.

44. SliusarenkoO, HeinritzJ, EmonetT, Jacobs-WagnerC (2011) High-throughput, subpixel precision analysis of bacterial morphogenesis and intracellular spatio-temporal dynamics. Mol Microbiol 80: 612–627.

Štítky
Genetika Reprodukčná medicína

Článok vyšiel v časopise

PLOS Genetics


2013 Číslo 5
Najčítanejšie tento týždeň
Najčítanejšie v tomto čísle
Kurzy

Zvýšte si kvalifikáciu online z pohodlia domova

Získaná hemofilie - Povědomí o nemoci a její diagnostika
nový kurz

Eozinofilní granulomatóza s polyangiitidou
Autori: doc. MUDr. Martina Doubková, Ph.D.

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#