Identification of the SlmA Active Site Responsible for Blocking Bacterial Cytokinetic Ring Assembly over the Chromosome

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Bacterial cells use chromosome-associated division inhibitors to help coordinate the processes of DNA replication and segregation with cytokinesis. SlmA from Escherichia coli, a member of the tetracycline repressor (TetR)–like protein family, is one example of this class of regulator. It blocks the assembly of the bacterial cytokinetic ring by interfering with the polymerization of the tubulin-like FtsZ protein in a manner that is dramatically stimulated upon specific DNA binding. Here we used a combination of molecular genetics and biochemistry to identify the active site of SlmA responsible for disrupting FtsZ polymerization. Interestingly, this site maps to a region of SlmA that in the published DNA–free structure is partially occluded by the DNA-binding domains. In this conformation, the SlmA structure resembles the drug/inducer-bound conformers of other TetR–like proteins, which in the absence of inducer require an inward rotation of their DNA-binding domains to bind successive major grooves on operator DNA. Our results are therefore consistent with a model in which DNA-binding activates SlmA by promoting a rotational movement of the DNA-binding domains that fully exposes the FtsZ-binding sites. SlmA may thus represent a special subclass of TetR–like proteins that have adapted conformational changes normally associated with inducer sensing in order to modulate an interaction with a partner protein. In this case, the adaptation ensures that SlmA only blocks cytokinesis in regions of the cell occupied by the origin-proximal portion of the chromosome where SlmA-binding sites are enriched.

Vyšlo v časopise: Identification of the SlmA Active Site Responsible for Blocking Bacterial Cytokinetic Ring Assembly over the Chromosome. PLoS Genet 9(2): e32767. doi:10.1371/journal.pgen.1003304
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pgen.1003304

Souhrn

Zdroje

1. de BoerPAJ (2010) Advances in understanding E. coli cell fission. Curr Opin Microbiol 13: 730–737 doi:10.1016/j.mib.2010.09.015.

2. BiEF, LutkenhausJ (1991) FtsZ ring structure associated with division in Escherichia coli. Nature 354: 161–164 doi:10.1038/354161a0.

3. Durand-HerediaJM, YuHH, De CarloS, LesserCF, JanakiramanA (2011) Identification and characterization of ZapC, a stabilizer of the FtsZ ring in Escherichia coli. J Bacteriol 193: 1405–1413 doi:10.1128/JB.01258-10.

4. HaleCA, ShiomiD, LiuB, BernhardtTG, MargolinW, et al. (2011) Identification of Escherichia coli ZapC (YcbW) as a component of the division apparatus that binds and bundles FtsZ polymers. J Bacteriol 193: 1393–1404 doi:10.1128/JB.01245-10.

5. Durand-HerediaJ, RivkinE, FanG, MoralesJ, JanakiramanA (2012) Identification of ZapD as a Cell Division Factor That Promotes the Assembly of FtsZ in Escherichia coli. J Bacteriol 194: 3189–3198 doi:10.1128/JB.00176-12.

6. AarsmanMEG, PietteA, FraipontC, VinkenvleugelTMF, Nguyen-DistècheM, et al. (2005) Maturation of the Escherichia coli divisome occurs in two steps. Mol Microbiol 55: 1631–1645 doi:10.1111/j.1365-2958.2005.04502.x.

7. de BoerPA, CrossleyRE, RothfieldLI (1989) A division inhibitor and a topological specificity factor coded for by the minicell locus determine proper placement of the division septum in E. coli. Cell 56: 641–649.

8. YuXC, MargolinW (1999) FtsZ ring clusters in min and partition mutants: role of both the Min system and the nucleoid in regulating FtsZ ring localization. Mol Microbiol 32: 315–326.

9. WoldringhCL, MulderE, HulsPG, VischerN (1991) Toporegulation of bacterial division according to the nucleoid occlusion model. Res Microbiol 142: 309–320.

10. LevinPA, ShimJJ, GrossmanAD (1998) Effect of minCD on FtsZ ring position and polar septation in Bacillus subtilis. J Bacteriol 180: 6048–6051.

11. WuLJ, ErringtonJ (2011) Nucleoid occlusion and bacterial cell division. Nat Rev Microbiol 10: 8–12 doi:10.1038/nrmicro2671.

12. LevinPA, MargolisPS, SetlowP, LosickR, SunD (1992) Identification of Bacillus subtilis genes for septum placement and shape determination. J Bacteriol 174: 6717–6728.

13. LutkenhausJ (2007) Assembly dynamics of the bacterial MinCDE system and spatial regulation of the Z ring. Annu Rev Biochem 76: 539–562 doi:10.1146/annurev.biochem.75.103004.142652.

14. ParkK-T, WuW, BattaileKP, LovellS, HolyoakT, et al. (2011) The Min oscillator uses MinD-dependent conformational changes in MinE to spatially regulate cytokinesis. Cell 146: 396–407 doi:10.1016/j.cell.2011.06.042.

15. RaskinDM, de BoerPA (1999) Rapid pole-to-pole oscillation of a protein required for directing division to the middle of Escherichia coli. Proc Natl Acad Sci USA 96: 4971–4976.

16. RaskinDM, de BoerPA (1999) MinDE-dependent pole-to-pole oscillation of division inhibitor MinC in Escherichia coli. J Bacteriol 181: 6419–6424.

17. HuZ, LutkenhausJ (1999) Topological regulation of cell division in Escherichia coli involves rapid pole to pole oscillation of the division inhibitor MinC under the control of MinD and MinE. Mol Microbiol 34: 82–90.

18. MarstonAL, ErringtonJ (1999) Selection of the midcell division site in Bacillus subtilis through MinD-dependent polar localization and activation of MinC. Mol Microbiol 33: 84–96.

19. BramkampM, EmminsR, WestonL, DonovanC, DanielRA, et al. (2008) A novel component of the division-site selection system of Bacillus subtilis and a new mode of action for the division inhibitor MinCD. Mol Microbiol 70: 1556–1569 doi:10.1111/j.1365-2958.2008.06501.x.

20. PatrickJE, KearnsDB (2008) MinJ (YvjD) is a topological determinant of cell division in Bacillus subtilis. Mol Microbiol 70: 1166–1179 doi:10.1111/j.1365-2958.2008.06469.x.

21. MeinhardtH, de BoerPA (2001) Pattern formation in Escherichia coli: a model for the pole-to-pole oscillations of Min proteins and the localization of the division site. Proc Natl Acad Sci USA 98: 14202–14207 doi:10.1073/pnas.251216598.

22. WuLJ, ErringtonJ (2004) Coordination of cell division and chromosome segregation by a nucleoid occlusion protein in Bacillus subtilis. Cell 117: 915–925 doi:10.1016/j.cell.2004.06.002.

23. BernhardtTG, de BoerPAJ (2005) SlmA, a nucleoid-associated, FtsZ binding protein required for blocking septal ring assembly over Chromosomes in E. coli. Mol Cell 18: 555–564 doi:10.1016/j.molcel.2005.04.012.

24. WuLJ, IshikawaS, KawaiY, OshimaT, OgasawaraN, et al. (2009) Noc protein binds to specific DNA sequences to coordinate cell division with chromosome segregation. EMBO J 28: 1940–1952 doi:10.1038/emboj.2009.144.

25. ChoH, McManusHR, DoveSL, BernhardtTG (2011) Nucleoid occlusion factor SlmA is a DNA-activated FtsZ polymerization antagonist. Proc Natl Acad Sci USA 108: 3773–3778 doi:10.1073/pnas.1018674108.

26. TonthatNK, AroldST, PickeringBF, Van DykeMW, LiangS, et al. (2011) Molecular mechanism by which the nucleoid occlusion factor, SlmA, keeps cytokinesis in check. EMBO J 30: 154–164 doi:10.1038/emboj.2010.288.

27. SchumacherMA, MillerMC, GrkovicS, BrownMH, SkurrayRA, et al. (2001) Structural mechanisms of QacR induction and multidrug recognition. Science 294: 2158–2163 doi:10.1126/science.1066020.

28. OrthP, SchnappingerD, HillenW, SaengerW, HinrichsW (2000) Structural basis of gene regulation by the tetracycline inducible Tet repressor-operator system. Nat Struct Biol 7: 215–219 doi:10.1038/73324.

29. MukherjeeA, LutkenhausJ (1998) Dynamic assembly of FtsZ regulated by GTP hydrolysis. EMBO J 17: 462–469 doi:10.1093/emboj/17.2.462.

30. KarimovaG, PidouxJ, UllmannA, LadantD (1998) A bacterial two-hybrid system based on a reconstituted signal transduction pathway. Proc Natl Acad Sci USA 95: 5752–5756.

31. EricksonHP, TaylorDW, TaylorKA, BramhillD (1996) Bacterial cell division protein FtsZ assembles into protofilament sheets and minirings, structural homologs of tubulin polymers. Proc Natl Acad Sci USA 93: 519–523.

32. MukherjeeA, LutkenhausJ (1994) Guanine nucleotide-dependent assembly of FtsZ into filaments. J Bacteriol 176: 2754–2758.

33. MukherjeeA, LutkenhausJ (1999) Analysis of FtsZ assembly by light scattering and determination of the role of divalent metal cations. J Bacteriol 181: 823–832.

34. YuXC, MargolinW (1997) Ca2+-mediated GTP-dependent dynamic assembly of bacterial cell division protein FtsZ into asters and polymer networks in vitro. EMBO J 16: 5455–5463 doi:10.1093/emboj/16.17.5455.

35. GonzálezJM, JiménezM, VélezM, MingoranceJ, AndreuJM, et al. (2003) Essential cell division protein FtsZ assembles into one monomer-thick ribbons under conditions resembling the crowded intracellular environment. J Biol Chem 278: 37664–37671 doi:10.1074/jbc.M305230200.

36. ShenB, LutkenhausJ (2010) Examination of the interaction between FtsZ and MinCN in E. coli suggests how MinC disrupts Z rings. Mol Microbiol 75: 1285–1298 doi:10.1111/j.1365-2958.2010.07055.x.

37. RamosJL, Martínez-BuenoM, Molina-HenaresAJ, TeránW, WatanabeK, et al. (2005) The TetR family of transcriptional repressors. Microbiol Mol Biol Rev 69: 326–356 doi:10.1128/MMBR.69.2.326-356.2005.

38. RouthMD, SuC-C, ZhangQ, YuEW (2009) Structures of AcrR and CmeR: insight into the mechanisms of transcriptional repression and multi-drug recognition in the TetR family of regulators. Biochim Biophys Acta 1794: 844–851 doi:10.1016/j.bbapap.2008.12.001.

39. Miller J (1972) Experiments in Molecular Genetics. New York: Cold Spring Harbor Laboratory.

40. JohnsonJE, LacknerLL, HaleCA, de BoerPAJ (2004) ZipA is required for targeting of DMinC/DicB, but not DMinC/MinD, complexes to septal ring assemblies in Escherichia coli. J Bacteriol 186: 2418–2429.

41. HaldimannA, WannerBL (2001) Conditional-replication, integration, excision, and retrieval plasmid-host systems for gene structure-function studies of bacteria. J Bacteriol 183: 6384–6393 doi:10.1128/JB.183.21.6384-6393.2001.

42. Gueiros-FilhoFJ, LosickR (2002) A widely conserved bacterial cell division protein that promotes assembly of the tubulin-like protein FtsZ. Genes Dev 16: 2544–2556 doi:10.1101/gad.1014102.