A Single Cohesin Complex Performs Mitotic and Meiotic Functions in the Protist

English version České info

The cohesion of sister chromatids in the interval between chromosome replication and anaphase is important for preventing the precocious separation, and hence nondisjunction, of chromatids. Cohesion is accomplished by a ring-shaped protein complex, cohesin; and its release at anaphase occurs when separase cleaves the complex's α-kleisin subunit. Cohesin has additional roles in facilitating DNA damage repair from the sister chromatid and in regulating gene expression. We tested the universality of the present model of cohesion by studying cohesin in the evolutionarily distant protist Tetrahymena thermophila. Localization of tagged cohesin components Smc1p and Rec8p (the α-kleisin) showed that cohesin is abundant in mitotic and meiotic nuclei. RNAi knockdown experiments demonstrated that cohesin is crucial for normal chromosome segregation and meiotic DSB repair. Unexpectedly, cohesin does not detach from chromosome arms in anaphase, yet chromosome segregation depends on the activity of separase (Esp1p). When Esp1p is depleted by RNAi, chromosomes become polytenic as they undergo multiple rounds of replication, but fail to separate. The cohesion of such bundles of numerous chromatids suggests that chromatids may be connected by factors in addition to topological linkage by cohesin rings. Although cohesin is not detected in transcriptionally active somatic nuclei, its loss causes a slight defect in their amitotic division. Notably, Tetrahymena uses a single version of α-kleisin for both mitosis and meiosis. Therefore, we propose that the differentiation of mitotic and meiotic cohesins found in most other model systems is not due to the need of a specialized meiotic cohesin, but due to additional roles of mitotic cohesin.

Vyšlo v časopise: A Single Cohesin Complex Performs Mitotic and Meiotic Functions in the Protist. PLoS Genet 9(3): e32767. doi:10.1371/journal.pgen.1003418
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pgen.1003418

Souhrn

Zdroje

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

2. SchleifferA, KaitnaS, Maurer-StrohS, GlotzerM, NasmythK, et al. (2003) Kleisins: a superfamily of bacterial and eukaryotic SMC protein partners. Mol Cell 11 : 571–575.

3. ZicklerD, KlecknerN (1999) Meiotic chromosomes: integrating structure and function. Annu Rev Genet 33 : 603–754.

4. San FilippoJ, SungP, KleinH (2008) Mechanism of eukaryotic homologous recombination. Annu Rev Biochem 77 : 229–257.

5. PetronczkiM, SiomosMF, NasmythK (2003) Un ménage à quatre: the molecular biology of chromosome segregation in meiosis. Cell 112 : 423–440.

6. PetersJ-M, TedeschiA, SchmitzJ (2008) The cohesin complex and its roles in chromosome biology. Genes Dev 22 : 3089–3114.

7. KleinF, MahrP, GalovaM, BuonomoSBC, MichaelisC, et al. (1999) A central role for cohesins in sister chromatid cohesion, formation of axial elements, and recombination during yeast meiosis. Cell 98 : 91–103.

8. PasierbekP, JantschM, MelcherM, SchleifferA, SchweizerD, et al. (2001) A Caenorhabditis elegans cohesion protein with functions in meiotic chromosome pairing and disjunction. Genes Dev 15 : 1349–1360.

9. SeversonAF, LingL, van ZuylenV, MeyerBJ (2009) The axial element protein HTP-3 promotes cohesin loading and meiotic axis assembly in C. elegans to implement the meiotic program of chromosome segregation. Genes Dev 23 : 1763–1778.

10. LiuZ, MakaroffCA (2006) Arabidopsis separase AESP is essential for embryo development and the release of cohesin during meiosis. Plant Cell 18 : 1213–1225.

11. EijpeM, OffenbergH, JessbergerR, RevenkovaE, HeytingC (2003) Meiotic cohesin REC8 marks the axial elements of rat synaptonemal complexes before cohesins SMC1ß and SMC3. J Cell Biol 160 : 657–670.

12. EllermeierC, SmithGR (2005) Cohesins are required for meiotic DNA breakage and recombination in Schizosaccharomyces pombe. Proc Natl Acad Sci USA 102 : 10952–10957.

13. WatrinE, PetersJ-M (2006) Cohesin and DNA damage repair. Exp Cell Res 312 : 2687–2693.

14. DorsettD, StrömL (2012) The ancient and evolving roles of cohesin in gene expression and DNA repair. Curr Biol 22: R240–R250.

15. LandeiraD, BartJ-M, Van TyneD, NavarroM (2009) Cohesin regulates VSG monoallelic expression in trypanosomes. J Cell Biol 186 : 243–254.

16. UhlmannF (2008) Molecular biology: cohesin branches out. Nature 451 : 777–778.

17. HadjurS, WilliamsLM, RyanNK, CobbBS, SextonT, et al. (2009) Cohesins form chromosomal cis-interactions at the developmentally regulated IFNG locus. Nature 460 : 410–413.

18. OriasE, CervantesMD, HamiltonE (2011) Tetrahymena thermophila, a unicellular eukaryote with separate germline and somatic genomes. Res Microbiol 162 : 578–586.

19. WolfeJ, HunterB, AdairWS (1976) A cytological study of micronuclear elongation during conjugation in Tetrahymena. Chromosoma 55 : 289–308.

20. RayCJr (1956) Meiosis and nuclear behaviour in Tetrahymena pyriformis. J Protozool 3 : 88–96.

21. SugaiT, HiwatashiK (1974) Cytologic and autoradiographic studies of the micronucleus at meiotic prophase in Tetrahymena pyriformis. J Protozool 21 : 542–548.

22. MartindaleDW, AllisCD, BrunsPJ (1982) Conjugation in Tetrahymena thermophila. A temporal analysis of cytological stages. Exp Cell Res 140 : 227–236.

23. LoidlJ, MochizukiK (2009) Tetrahymena meiotic nuclear reorganization is induced by a checkpoint kinase-dependent response to DNA damage. Mol Biol Cell 20 : 2428–2437.

24. EisenJA, CoyneRS, WuM, WuD, ThiagarajanM, et al. (2006) Macronuclear genome sequence of the ciliate Tetrahymena thermophila, a model eukaryote. PLoS Biol 4: e286 doi:10.1371/journal.pbio.0040286.

25. HaeringCH, SchoffneggerD, NishinoT, HelmhartW, NasmythK, et al. (2004) Structure and stability of cohesin's Smc1-kleisin interaction. Mol Cell 15 : 951–964.

26. MiaoW, XiongJ, BowenJ, WangW, LiuY, et al. (2009) Microarray analyses of gene expression during the Tetrahymena thermophila life cycle. PLoS ONE 4: e4429 doi:10.1371/journal.pone.0004429.

27. BuonomoSBC, ClyneRK, FuchsJ, LoidlJ, UhlmannF, et al. (2000) Disjunction of homologous chromosomes in meiosis I depends on proteolytic cleavage of the meiotic cohesin Rec8p by separin. Cell 103 : 387–398.

28. KitajimaTS, KawashimaSA, WatanabeY (2004) The konserved kinetochore protein Shugoshin protects centromeric cohesion during meiosis. Nature 427 : 510–517.

29. ZenvirthD, LoidlJ, KleinS, ArbelA, ShemeshR, et al. (1997) Switching yeast from meiosis to mitosis: double-strand break repair, recombination and synaptonemal complex. Genes Cells 2 : 487–498.

30. CervantesMD, CoyneRS, XiXH, YaoMC (2006) The condensin complex is essential for amitotic segregation of bulk chromosomes, but not nucleoli, in the ciliate Tetrahymena thermophila. Mol Cell Biol 26 : 4690–4700.

31. McDonaldBB (1966) The exchange of RNA and protein during conjugation in Tetrahymena. J Protozool 13 : 277–285.

32. WaizeneggerIC, HaufS, MeinkeA, PetersJ-M (2000) Two distinct pathways remove mammalian cohesin from chromosome arms in prophase and from centromeres in anaphase. Cell 103 : 399–410.

33. CaiX, DongFG, EdelmannRE, MakaroffCA (2003) The Arabidopsis SYN1 cohesin protein is required for sister chromatid arm cohesion and homologous chromosome pairing. J Cell Sci 116 : 2999–3007.

34. KitajimaTS, YokobayashiS, YamamotoM, WatanabeY (2003) Distinct cohesin complexes organize meiotic chromosome domains. Science 300 : 1152–1155.

35. LosadaA, HiranoM, HiranoT (2002) Cohesin release is required for sister chromatid resolution, but not for condensin-mediated compaction, at the onset of mitosis. Genes Dev 16 : 3004–3016.

36. KudoNR, WassmannK, AngerM, SchuhM, WirthKG, et al. (2006) Resolution of chiasmata in oocytes requires separase-mediated proteolysis. Cell 126 : 135–146.

37. TomonagaT, NagaoK, KawasakiY, FuruyaK, MurakamiA, et al. (2000) Characterization of fission yeast cohesin: essential anaphase proteolysis of Rad21 phosphorylated in the S phase. Genes Dev 14 : 2757–2770.

38. Howard-TillRA, LukaszewiczA, LoidlJ (2011) The recombinases Rad51 and Dmc1 play distinct roles in DNA break repair and recombination partner choice in the meiosis of Tetrahymena. PLoS Genet 7: e1001359 doi:10.1371/journal.pgen.1001359.

39. WoodardJ, KaneshiroE, GorovskyMA (1972) Cytochemical studies on the problem of macronuclear subnuclei in Tetrahymena. Genetics 70 : 251–260.

40. JessbergerR (2009) Cohesin's dual role in the DNA damage response: repair and checkpoint activation. EMBO J 28 : 2491–2493.

41. StrömL, LindroosHB, ShirahigeK, SjögrenC (2004) Postreplicative recruitment of cohesin to double-strand breaks is required for DNA repair. Mol Cell 16 : 1003–1015.

42. ÜnalE, Arbel-EdenA, SattlerU, ShroffR, LichtenM, et al. (2004) DNA damage response pathway uses histone modification to assemble a double-strand break-specific cohesin domain. Mol Cell 16 : 991–1002.

43. CampbellC, RomeroDP (1998) Identification and characterization of the RAD51 gene from the ciliate Tetrahymena thermophila. Nucleic Acids Res 26 : 3165–3172.

44. LukaszewiczA, Howard-TillRA, NovatchkovaM, MochizukiK, LoidlJ (2010) MRE11 and COM1/SAE2 are required for double-strand break repair and efficient chromosome pairing during meiosis of the protist Tetrahymena. Chromosoma 119 : 505–518.

45. Howard-TillRA, YaoMC (2006) Induction of gene silencing by hairpin RNA expression in Tetrahymena thermophila reveals a second small RNA pathway. Mol Cell Biol 26 : 8731–8742.

46. CleffmanG (1980) Chromatin elimination and the genetic organisation of the macronucleus in Tetrahymena thermophila. Chromosoma 78 : 313–325.

47. WeiY, MizzenCA, CookRG, GorovskyMA, AllisCD (1998) Phosphorylation of histone H3 at serine 10 is correlated with chromosome condensation during mitosis and meiosis in Tetrahymena. Proc Natl Acad Sci USA 95 : 7480–7484.

48. BaudrimontA, PenknerA, WoglarA, MamnunYM, HulekM, et al. (2011) A new thermosensitive smc-3 allele reveals involvement of cohesin in homologous recombination in C. elegans. PLoS ONE 6: e24799 doi:10.1371/journal.pone.0024799.

49. Heidinger-PauliJM, MertO, DavenportC, GuacciV, KoshlandD (2010) Systematic reduction of cohesin differentially affects chromosome segregation, condensation, and DNA repair. Curr Biol 20 : 957–963.

50. BrarGA, HochwagenA, EeLS, AmonA (2009) The multiple roles of cohesin in meiotic chromosome morphogenesis and pairing. Mol Biol Cell 20 : 1030–1047.

51. LightfootJ, TestoriS, BarrosoC, Martinez-PerezE (2011) Loading of meiotic cohesin by SCC-2 is required for early processing of DSBs and for the DNA damage checkpoint. Curr Biol 21 : 1421–1430.

52. FarcasA-M, UluocakP, HelmhartW, NasmythK (2011) Cohesin's concatenation of sister DNAs maintains their intertwining. Mol Cell 44 : 97–107.

53. ShimadaK, GasserSM (2007) The origin recognition complex functions in sister-chromatid cohesion in Saccharomyces cerevisiae. Cell 128 : 85–99.

54. GuacciV, KoshlandD (2012) Cohesin-independent segregation of sister chromatids in budding yeast. Mol Biol Cell 23 : 729–739.

55. BrunsPJ, BrussardTEB (1981) Nullisomic Tetrahymena: eliminating germinal chromosomes. Science 213 : 549–551.

56. Frankel J (2000) Cell biology of Tetrahymena thermophila. In: Asai DJ, Forney JD, editors. Tetrahymena thermophila. San Diego: Academic Press. pp. 27–125.

57. KimKP, WeinerBM, ZhangL, JordanA, DekkerJ, et al. (2010) Sister cohesion and structural axis components mediate homolog bias of meiotic recombination. Cell 143 : 924–937.

58. KugouK, FukudaT, YamadaS, ItoM, SasanumaH, et al. (2009) Rec8 guides canonical Spo11 distribution along yeast meiotic chromosomes. Mol Biol Cell 20 : 3064–3076.

59. HiranoT (2005) Cell biology: holding sisters for repair. Nature 433 : 467–468.

60. Cole E, Sugai T (2012) Developmental progression of Tetrahymena through the cell cycle and conjugation. In: Collins K, editors. Tetrahymena thermophila. San Diego: Academic Press. pp. 177–236.

61. WirthKG, WutzG, KudoNR, DesdouetsC, ZetterbergA, et al. (2006) Separase: a universal trigger for sister chromatid disjunction but not chromosome cycle progression. J Cell Biol 172 : 847–860.

62. KhetaniRS, BickelSE (2007) Regulation of meiotic cohesion and chromosome core morphogenesis during pachytene in Drosophila oocytes. J Cell Sci 120 : 3123–3137.

63. LeeJ, OgushiS, SaitouM, HiranoT (2011) Condensins I and II are essential for construction of bivalent chromosomes in mouse oocytes. Mol Biol Cell 22 : 3465–3477.

64. HerranY, Gutierrez-CaballeroC, Sanchez-MartinM, HernandezT, VieraA, et al. (2011) The cohesin subunit RAD21L functions in meiotic synapsis and exhibits sexual dimorphism in fertility. EMBO J 30 : 3091–3105.

65. IshiguroK, KimJ, Fujiyama-NakamuraS, KatoS, WatanabeY (2011) A new meiosis-specific cohesin complex implicated in the cohesin code for homologous pairing. EMBO Rep 12 : 267–275.

66. McDonaldBB (1962) Synthesis of deoxyribonucleic acid by micro -⁠ and macronuclei of Tetrahymena pyriformis. J Cell Biol 13 : 193–203.

67. DoerderFP, De BaultLE (1975) Cytofluorometric analysis of nuclear DNA during meiosis, fertilization and macronuclear development in the ciliate Tetrahymena pyriformis, syngen 1. J Cell Sci 17 : 471–493.

68. Heidinger-PauliJM, ÜnalE, GuacciV, KoshlandD (2008) The kleisin subunit of cohesin dictates damage-induced cohesion. Mol Cell 31 : 47–56.

69. Orias E, Hamilton EP, Orias JD (2000) Tetrahymena as a laboratory organism: useful strains, cell culture, and cell line maintenance. In: Asai DJ, Forney JD, editors. Tetrahymena thermophila. San Diego: Academic Press. pp. 189–211.

70. Cassidy-HanleyD, BowenJ, LeeJH, ColeE, VerPlankLA, et al. (1997) Germline and somatic transformation of mating Tetrahymena termophila by particle bombardment. Genetics 146 : 135–147.

71. KöcherT, PichlerP, SwartR, MechtlerK (2012) Analysis of protein mixtures from whole-cell extracts by single-run nanoLC-MS/MS using ultralong gradients. Nature Protoc 7 : 882–890.

72. LoidlJ, ScherthanH (2004) Organization and pairing of meiotic chromosomes in the ciliate Tetrahymena thermophila. J Cell Sci 117 : 5791–5801.

73. MochizukiK, NovatchkovaM, LoidlJ (2008) DNA double-strand breaks, but not crossovers, are required for the reorganization of meiotic nuclei in Tetrahymena. J Cell Sci 121 : 2148–2158.

74. RonquistF, TeslenkoM, van der MarkP, AyresDL, DarlingA, et al. (2012) MrBayes 3.2: efficient bayesian phylogenetic inference and model choice across a large model space. Syst Biol Epub ahead of print.