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Cohesin Rings Devoid of Scc3 and Pds5 Maintain Their Stable Association with the DNA


Cohesin is a protein complex that forms a ring around sister chromatids thus holding them together. The ring is composed of three proteins: Smc1, Smc3 and Scc1. The roles of three additional proteins that associate with the ring, Scc3, Pds5 and Wpl1, are not well understood. It has been proposed that these three factors form a complex that stabilizes the ring and prevents it from opening. This activity promotes sister chromatid cohesion but at the same time poses an obstacle for the initial entrapment of sister DNAs. This hindrance to cohesion establishment is overcome during DNA replication via acetylation of the Smc3 subunit by the Eco1 acetyltransferase. However, the full mechanistic consequences of Smc3 acetylation remain unknown. In the current work, we test the requirement of Scc3 and Pds5 for the stable association of cohesin with DNA. We investigated the consequences of Scc3 and Pds5 depletion in vivo using degron tagging in budding yeast. The previously described DHFR–based N-terminal degron as well as a novel Eco1-derived C-terminal degron were employed in our study. Scc3 and Pds5 associate with cohesin complexes independently of each other and require the Scc1 “core” subunit for their association with chromosomes. Contrary to previous data for Scc1 downregulation, depletion of either Scc3 or Pds5 had a strong effect on sister chromatid cohesion but not on cohesin binding to DNA. Quantity, stability and genome-wide distribution of cohesin complexes remained mostly unchanged after the depletion of Scc3 and Pds5. Our findings are inconsistent with a previously proposed model that Scc3 and Pds5 are cohesin maintenance factors required for cohesin ring stability or for maintaining its association with DNA. We propose that Scc3 and Pds5 specifically function during cohesion establishment in S phase.


Vyšlo v časopise: Cohesin Rings Devoid of Scc3 and Pds5 Maintain Their Stable Association with the DNA. PLoS Genet 8(8): e32767. doi:10.1371/journal.pgen.1002856
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pgen.1002856

Souhrn

Cohesin is a protein complex that forms a ring around sister chromatids thus holding them together. The ring is composed of three proteins: Smc1, Smc3 and Scc1. The roles of three additional proteins that associate with the ring, Scc3, Pds5 and Wpl1, are not well understood. It has been proposed that these three factors form a complex that stabilizes the ring and prevents it from opening. This activity promotes sister chromatid cohesion but at the same time poses an obstacle for the initial entrapment of sister DNAs. This hindrance to cohesion establishment is overcome during DNA replication via acetylation of the Smc3 subunit by the Eco1 acetyltransferase. However, the full mechanistic consequences of Smc3 acetylation remain unknown. In the current work, we test the requirement of Scc3 and Pds5 for the stable association of cohesin with DNA. We investigated the consequences of Scc3 and Pds5 depletion in vivo using degron tagging in budding yeast. The previously described DHFR–based N-terminal degron as well as a novel Eco1-derived C-terminal degron were employed in our study. Scc3 and Pds5 associate with cohesin complexes independently of each other and require the Scc1 “core” subunit for their association with chromosomes. Contrary to previous data for Scc1 downregulation, depletion of either Scc3 or Pds5 had a strong effect on sister chromatid cohesion but not on cohesin binding to DNA. Quantity, stability and genome-wide distribution of cohesin complexes remained mostly unchanged after the depletion of Scc3 and Pds5. Our findings are inconsistent with a previously proposed model that Scc3 and Pds5 are cohesin maintenance factors required for cohesin ring stability or for maintaining its association with DNA. We propose that Scc3 and Pds5 specifically function during cohesion establishment in S phase.


Zdroje

1. NasmythK, HaeringCH (2009) Cohesin: its roles and mechanisms. Annu Rev Genet 43: 525–558.

2. HaeringCH, FarcasAM, ArumugamP, MetsonJ, NasmythK (2008) The cohesin ring concatenates sister DNA molecules. Nature 454: 297–301.

3. OnnI, Heidinger-PauliJM, GuacciV, UnalE, KoshlandDE (2008) Sister chromatid cohesion: a simple concept with a complex reality. Annu Rev Cell Dev Biol 24: 105–129.

4. HaeringCH, SchoffneggerD, NishinoT, HelmhartW, NasmythK, et al. (2004) Structure and Stability of Cohesin's Smc1-Kleisin Interaction. Mol Cell 15: 951–964.

5. SkibbensRV, CorsonLB, KoshlandD, HieterP (1999) Ctf7p is essential for sister chromatid cohesion and links mitotic chromosome structure to the DNA replication machinery. Genes Dev 13: 307–319.

6. TothA, CioskR, UhlmannF, GalovaM, SchleifferA, et al. (1999) Yeast cohesin complex requires a conserved protein, Eco1p(Ctf7), to establish cohesion between sister chromatids during DNA replication. Genes Dev 13: 320–333.

7. IvanovD, SchleifferA, EisenhaberF, MechtlerK, HaeringCH, et al. (2002) Eco1 is a novel acetyltransferase that can acetylate proteins involved in cohesion. Curr Biol 12: 323–328.

8. ZhangJ, ShiX, LiY, KimBJ, JiaJ, et al. (2008) Acetylation of Smc3 by Eco1 is required for S phase sister chromatid cohesion in both human and yeast. Mol Cell 31: 143–151.

9. Rolef Ben-ShaharT, HeegerS, LehaneC, EastP, FlynnH, et al. (2008) Eco1-dependent cohesin acetylation during establishment of sister chromatid cohesion. Science 321: 563–566.

10. UnalE, Heidinger-PauliJM, KimW, GuacciV, OnnI, et al. (2008) A molecular determinant for the establishment of sister chromatid cohesion. Science 321: 566–569.

11. Heidinger-PauliJM, UnalE, KoshlandD (2009) Distinct targets of the Eco1 acetyltransferase modulate cohesion in S phase and in response to DNA damage. Mol Cell 34: 311–321.

12. BeckouetF, HuB, RoigMB, SutaniT, KomataM, et al. (2011) An Smc3 acetylation cycle is essential for establishment of sister chromatid cohesion. Mol Cell 39: 689–699.

13. BorgesV, LehaneC, Lopez-SerraL, FlynnH, SkehelM, et al. (2011) Hos1 deacetylates Smc3 to close the cohesin acetylation cycle. Mol Cell 39: 677–688.

14. XiongB, LuS, GertonJL (2011) Hos1 is a lysine deacetylase for the smc3 subunit of cohesin. Curr Biol 20: 1660–1665.

15. SutaniT, KawaguchiT, KannoR, ItohT, ShirahigeK (2009) Budding yeast Wpl1(Rad61)-Pds5 complex counteracts sister chromatid cohesion-establishing reaction. Curr Biol 19: 492–497.

16. KuengS, HegemannB, PetersBH, LippJJ, SchleifferA, et al. (2006) Wapl controls the dynamic association of cohesin with chromatin. Cell 127: 955–967.

17. GandhiR, GillespiePJ, HiranoT (2006) Human Wapl is a cohesin-binding protein that promotes sister-chromatid resolution in mitotic prophase. Curr Biol 16: 2406–2417.

18. RowlandBD, RoigMB, NishinoT, KurzeA, UluocakP, et al. (2009) Building sister chromatid cohesion: smc3 acetylation counteracts an antiestablishment activity. Mol Cell 33: 763–774.

19. PanizzaS, TanakaT, HochwagenA, EisenhaberF, NasmythK (2000) Pds5 cooperates with cohesin in maintaining sister chromatid cohesion. Curr Biol 10: 1557–1564.

20. HartmanT, SteadK, KoshlandD, GuacciV (2000) Pds5p is an essential chromosomal protein required for both sister chromatid cohesion and condensation in Saccharomyces cerevisiae. J Cell Biol 151: 613–626.

21. SumaraI, VorlauferE, GieffersC, PetersBH, PetersJM (2000) Characterization of vertebrate cohesin complexes and their regulation in prophase. J Cell Biol 151: 749–762.

22. LosadaA, YokochiT, KobayashiR, HiranoT (2000) Identification and characterization of SA/Scc3p subunits in the Xenopus and human cohesin complexes. J Cell Biol 150: 405–416.

23. LosadaA, YokochiT, HiranoT (2005) Functional contribution of Pds5 to cohesin-mediated cohesion in human cells and Xenopus egg extracts. J Cell Sci 118: 2133–2141.

24. TanakaK, HaoZ, KaiM, OkayamaH (2001) Establishment and maintenance of sister chromatid cohesion in fission yeast by a unique mechanism. Embo J 20: 5779–5790.

25. NeuwaldAF, HiranoT (2000) HEAT repeats associated with condensins, cohesins, and other complexes involved in chromosome-related functions. Genome Res 10: 1445–1452.

26. HaufS, RoitingerE, KochB, DittrichCM, MechtlerK, et al. (2005) Dissociation of cohesin from chromosome arms and loss of arm cohesion during early mitosis depends on phosphorylation of SA2. PLoS Biol 3: e69.

27. BernardP, SchmidtCK, VaurS, DheurS, DrogatJ, et al. (2008) Cell-cycle regulation of cohesin stability along fission yeast chromosomes. Embo J 27: 111–121.

28. HaeringCH, LoweJ, HochwagenA, NasmythK (2002) Molecular architecture of SMC proteins and the yeast cohesin complex. Mol Cell 9: 773–788.

29. ShintomiK, HiranoT (2009) Releasing cohesin from chromosome arms in early mitosis: opposing actions of Wapl-Pds5 and Sgo1. Genes Dev 23: 2224–2236.

30. HuB, ItohT, MishraA, KatohY, ChanKL, et al. (2011) ATP hydrolysis is required for relocating cohesin from sites occupied by its Scc2/4 loading complex. Curr Biol 21: 12–24.

31. GruberS, ArumugamP, KatouY, KuglitschD, HelmhartW, et al. (2006) Evidence that Loading of Cohesin Onto Chromosomes Involves Opening of Its SMC Hinge. Cell 127: 523–537.

32. KanemakiM, Sanchez-DiazA, GambusA, LabibK (2003) Functional proteomic identification of DNA replication proteins by induced proteolysis in vivo. Nature 423: 720–724.

33. Sanchez-DiazA, KanemakiM, MarchesiV, LabibK (2004) Rapid depletion of budding yeast proteins by fusion to a heat-inducible degron. Sci STKE 2004: PL8.

34. YenK, GitshamP, WishartJ, OliverSG, ZhangN (2003) An improved tetO promoter replacement system for regulating the expression of yeast genes. Yeast 20: 1255–1262.

35. LyonsNA, MorganDO (2011) Cdk1-dependent destruction of Eco1 prevents cohesion establishment after S phase. Mol Cell 42: 378–389.

36. WeitzerS, LehaneC, UhlmannF (2003) A model for ATP hydrolysis-dependent binding of cohesin to DNA. Curr Biol 13: 1930–1940.

37. YehE, HaaseJ, PaliulisLV, JoglekarA, BondL, et al. (2008) Pericentric chromatin is organized into an intramolecular loop in mitosis. Curr Biol 18: 81–90.

38. IvanovD, NasmythK (2005) A topological interaction between cohesin rings and a circular minichromosome. Cell 122: 849–860.

39. Ben-ShaharTR, HeegerS, LehaneC, EastP, FlynnH, et al. (2008) Eco1-dependent cohesin acetylation during establishment of sister chromatid cohesion. Science 321: 563–566.

40. FerniusJ, HardwickKG (2007) Bub1 kinase targets Sgo1 to ensure efficient chromosome biorientation in budding yeast mitosis. PLoS Genet 3: e213.

41. WarrenCD, EckleyDM, LeeMS, HannaJS, HughesA, et al. (2004) S-phase checkpoint genes safeguard high-fidelity sister chromatid cohesion. Mol Biol Cell 15: 1724–1735.

42. GameJC, BirrellGW, BrownJA, ShibataT, BaccariC, et al. (2003) Use of a genome-wide approach to identify new genes that control resistance of Saccharomyces cerevisiae to ionizing radiation. Radiat Res 160: 14–24.

43. SjogrenC, NasmythK (2001) Sister chromatid cohesion is required for postreplicative double-strand break repair in Saccharomyces cerevisiae. Curr Biol 11: 991–995.

44. WatrinE, PetersJM (2009) The cohesin complex is required for the DNA damage-induced G2/M checkpoint in mammalian cells. Embo J 28: 2625–2635.

45. LisbyM, MortensenUH, RothsteinR (2003) Colocalization of multiple DNA double-strand breaks at a single Rad52 repair centre. Nat Cell Biol 5: 572–577.

46. Heidinger-PauliJM, MertO, DavenportC, GuacciV, KoshlandD (2011) Systematic reduction of cohesin differentially affects chromosome segregation, condensation, and DNA repair. Curr Biol 20: 957–963.

47. GruberS, HaeringCH, NasmythK (2003) Chromosomal cohesin forms a ring. Cell 112: 765–777.

48. GerlichD, KochB, DupeuxF, PetersJM, EllenbergJ (2006) Live-cell imaging reveals a stable cohesin-chromatin interaction after but not before DNA replication. Curr Biol 16: 1571–1578.

49. VassS, CotterillS, ValdeolmillosAM, BarberoJL, LinE, et al. (2003) Depletion of Drad21/Scc1 in Drosophila cells leads to instability of the cohesin complex and disruption of mitotic progression. Curr Biol 13: 208–218.

50. NonakaN, KitajimaT, YokobayashiS, XiaoG, YamamotoM, et al. (2002) Recruitment of cohesin to heterochromatic regions by Swi6/HP1 in fission yeast. Nat Cell Biol 4: 89–93.

51. RubioED, ReissDJ, WelcshPL, DistecheCM, FilippovaGN, et al. (2008) CTCF physically links cohesin to chromatin. Proc Natl Acad Sci U S A 105: 8309–8314.

52. Mc IntyreJ, MullerEG, WeitzerS, SnydsmanBE, DavisTN, et al. (2007) In vivo analysis of cohesin architecture using FRET in the budding yeast Saccharomyces cerevisiae. Embo J 26: 3783–3793.

53. JankeC, MagieraMM, RathfelderN, TaxisC, ReberS, et al. (2004) A versatile toolbox for PCR-based tagging of yeast genes: new fluorescent proteins, more markers and promoter substitution cassettes. Yeast 21: 947–962.

54. StudierFW (2005) Protein production by auto-induction in high density shaking cultures. Protein Expr Purif 41: 207–234.

55. IvanovD, KwakYT, NeeE, GuoJ, Garcia-MartinezLF, et al. (1999) Cyclin T1 domains involved in complex formation with Tat and TAR RNA are critical for tat-activation. J Mol Biol 288: 41–56.

56. IvanovD, NasmythK (2007) A physical assay for sister chromatid cohesion in vitro. Mol Cell 27: 300–310.

57. TanakaT, CosmaMP, WirthK, NasmythK (1999) Identification of cohesin association sites at centromeres and along chromosome arms. Cell 98: 847–858.

58. OssowskiS, SchneebergerK, ClarkRM, LanzC, WarthmannN, et al. (2008) Sequencing of natural strains of Arabidopsis thaliana with short reads. Genome Res 18: 2024–2033.

59. MichaelisC, CioskR, NasmythK (1997) Cohesins: chromosomal proteins that prevent premature separation of sister chromatids. Cell 91: 35–45.

60. LiangC, StillmanB (1997) Persistent initiation of DNA replication and chromatin-bound MCM proteins during the cell cycle in cdc6 mutants. Genes Dev 11: 3375–3386.

61. ZhangN, KuznetsovSG, SharanSK, LiK, RaoPH, et al. (2008) A handcuff model for the cohesin complex. J Cell Biol 183: 1019–1031.

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