Budding Yeast Greatwall and Endosulfines Control Activity and Spatial Regulation of PP2A for Timely Mitotic Progression

English version České info

Entry into mitosis is triggered by cyclinB/Cdk1, whose activity is abruptly raised by a positive feedback loop. The Greatwall kinase phosphorylates proteins of the endosulfine family and allows them to bind and inhibit the main Cdk1-counteracting PP2A-B55 phosphatase, thereby promoting mitotic entry. In contrast to most eukaryotic systems, Cdc14 is the main Cdk1-antagonizing phosphatase in budding yeast, while the PP2A^Cdc55 phosphatase promotes, instead of preventing, mitotic entry by participating to the positive feedback loop of Cdk1 activation. Here we show that budding yeast endosulfines (Igo1 and Igo2) bind to PP2A^Cdc55 in a cell cycle-regulated manner upon Greatwall (Rim15)-dependent phosphorylation. Phosphorylated Igo1 inhibits PP2A^Cdc55 activity in vitro and induces mitotic entry in Xenopus egg extracts, indicating that it bears a conserved PP2A-binding and -inhibitory activity. Surprisingly, deletion of IGO1 and IGO2 in yeast cells leads to a decrease in PP2A phosphatase activity, suggesting that endosulfines act also as positive regulators of PP2A in yeast. Consistently, RIM15 and IGO1/2 promote, like PP2A^Cdc55, timely entry into mitosis under temperature-stress, owing to the accumulation of Tyr-phosphorylated Cdk1. In addition, they contribute to the nuclear export of PP2A^Cdc55, which has recently been proposed to promote mitotic entry. Altogether, our data indicate that Igo proteins participate in the positive feedback loop for Cdk1 activation. We conclude that Greatwall, endosulfines, and PP2A are part of a regulatory module that has been conserved during evolution irrespective of PP2A function in the control of mitosis. However, this conserved module is adapted to account for differences in the regulation of mitotic entry in different organisms.

Vyšlo v časopise: Budding Yeast Greatwall and Endosulfines Control Activity and Spatial Regulation of PP2A for Timely Mitotic Progression. PLoS Genet 9(7): e32767. doi:10.1371/journal.pgen.1003575
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pgen.1003575

Souhrn

Zdroje

1. MorganDO (1995) Principles of CDK regulation. Nature 374 : 131–134.

2. AbrieuA, BrassacT, GalasS, FisherD, LabbeJC, et al. (1998) The Polo-like kinase Plx1 is a component of the MPF amplification loop at the G2/M-phase transition of the cell cycle in Xenopus eggs. J Cell Sci 111(Pt 12): 1751–1757.

3. MortensenEM, HaasW, GygiM, GygiSP, KelloggDR (2005) Cdc28-dependent regulation of the Cdc5/Polo kinase. Curr Biol 15 : 2033–2037.

4. KumagaiA, DunphyWG (1996) Purification and molecular cloning of Plx1, a Cdc25-regulatory kinase from Xenopus egg extracts. Science 273 : 1377–1380.

5. QianYW, EriksonE, TaiebFE, MallerJL (2001) The polo-like kinase Plx1 is required for activation of the phosphatase Cdc25C and cyclin B-Cdc2 in Xenopus oocytes. Mol Biol Cell 12 : 1791–1799.

6. HarveySL, CharletA, HaasW, GygiSP, KelloggDR (2005) Cdk1-dependent regulation of the mitotic inhibitor Wee1. Cell 122 : 407–420.

7. MuellerPR, ColemanTR, DunphyWG (1995) Cell cycle regulation of a Xenopus Wee1-like kinase. Mol Biol Cell 6 : 119–134.

8. TangZ, ColemanTR, DunphyWG (1993) Two distinct mechanisms for negative regulation of the Wee1 protein kinase. EMBO J 12 : 3427–3436.

9. WatanabeN, AraiH, IwasakiJ, ShiinaM, OgataK, et al. (2005) Cyclin-dependent kinase (CDK) phosphorylation destabilizes somatic Wee1 via multiple pathways. Proc Natl Acad Sci U S A 102 : 11663–11668.

10. LindqvistA, Rodriguez-BravoV, MedemaRH (2009) The decision to enter mitosis: feedback and redundancy in the mitotic entry network. J Cell Biol 185 : 193–202.

11. PomereningJR, SontagED, FerrellJEJr (2003) Building a cell cycle oscillator: hysteresis and bistability in the activation of Cdc2. Nat Cell Biol 5 : 346–351.

12. BahlerJ (2005) Cell-cycle control of gene expression in budding and fission yeast. Annu Rev Genet 39 : 69–94.

13. AsanoS, ParkJE, SakchaisriK, YuLR, SongS, et al. (2005) Concerted mechanism of Swe1/Wee1 regulation by multiple kinases in budding yeast. EMBO J 24 : 2194–2204.

14. GrayCH, GoodVM, TonksNK, BarfordD (2003) The structure of the cell cycle protein Cdc14 reveals a proline-directed protein phosphatase. Embo J 22 : 3524–3535.

15. VisintinR, CraigK, HwangES, PrinzS, TyersM, et al. (1998) The phosphatase Cdc14 triggers mitotic exit by reversal of Cdk -⁠ dependent phosphorylation. Mol Cell 2 : 709–718.

16. ShouW, SeolJH, ShevchenkoA, BaskervilleC, MoazedD, et al. (1999) Exit from mitosis is triggered by Tem1-dependent release of the protein phosphatase Cdc14 from nucleolar RENT complex. Cell 97 : 233–244.

17. VisintinR, HwangES, AmonA (1999) Cfi1 prevents premature exit from mitosis by anchoring Cdc14 phosphatase in the nucleolus [see comments]. Nature 398 : 818–823.

18. WurzenbergerC, GerlichDW (2011) Phosphatases: providing safe passage through mitotic exit. Nat Rev Mol Cell Biol 12 : 469–482.

19. Mayer-JaekelRE, HemmingsBA (1994) Protein phosphatase 2A–a ‘menage a trois’. Trends Cell Biol 4 : 287–291.

20. JiangY (2006) Regulation of the cell cycle by protein phosphatase 2A in Saccharomyces cerevisiae. Microbiol Mol Biol Rev 70 : 440–449.

21. Mayer-JaekelRE, OhkuraH, FerrignoP, AndjelkovicN, ShiomiK, et al. (1994) Drosophila mutants in the 55 kDa regulatory subunit of protein phosphatase 2A show strongly reduced ability to dephosphorylate substrates of p34cdc2. J Cell Sci 107(Pt 9): 2609–2616.

22. MochidaS, IkeoS, GannonJ, HuntT (2009) Regulated activity of PP2A-B55 delta is crucial for controlling entry into and exit from mitosis in Xenopus egg extracts. EMBO J 28 : 2777–2785.

23. CastilhoPV, WilliamsBC, MochidaS, ZhaoY, GoldbergML (2009) The M phase kinase Greatwall (Gwl) promotes inactivation of PP2A/B55delta, a phosphatase directed against CDK phosphosites. Mol Biol Cell 20 : 4777–4789.

24. SchmitzMH, HeldM, JanssensV, HutchinsJR, HudeczO, et al. (2010) Live-cell imaging RNAi screen identifies PP2A-B55alpha and importin-beta1 as key mitotic exit regulators in human cells. Nat Cell Biol 12 : 886–893.

25. LorcaT, BernisC, VigneronS, BurgessA, BrioudesE, et al. (2010) Constant regulation of both the MPF amplification loop and the Greatwall-PP2A pathway is required for metaphase II arrest and correct entry into the first embryonic cell cycle. J Cell Sci 123 : 2281–2291.

26. VigneronS, BrioudesE, BurgessA, LabbeJC, LorcaT, et al. (2009) Greatwall maintains mitosis through regulation of PP2A. EMBO J 28 : 2786–2793.

27. YuJ, ZhaoY, LiZ, GalasS, GoldbergML (2006) Greatwall kinase participates in the Cdc2 autoregulatory loop in Xenopus egg extracts. Mol Cell 22 : 83–91.

28. Bettencourt-DiasM, GietR, SinkaR, MazumdarA, LockWG, et al. (2004) Genome-wide survey of protein kinases required for cell cycle progression. Nature 432 : 980–987.

29. YuJ, FlemingSL, WilliamsB, WilliamsEV, LiZ, et al. (2004) Greatwall kinase: a nuclear protein required for proper chromosome condensation and mitotic progression in Drosophila. J Cell Biol 164 : 487–492.

30. Gharbi-AyachiA, LabbeJC, BurgessA, VigneronS, StrubJM, et al. (2010) The substrate of Greatwall kinase, Arpp19, controls mitosis by inhibiting protein phosphatase 2A. Science 330 : 1673–1677.

31. MochidaS, MaslenSL, SkehelM, HuntT (2010) Greatwall phosphorylates an inhibitor of protein phosphatase 2A that is essential for mitosis. Science 330 : 1670–1673.

32. RangoneH, WegelE, GattMK, YeungE, FlowersA, et al. (2011) Suppression of scant identifies Endos as a substrate of greatwall kinase and a negative regulator of protein phosphatase 2A in mitosis. PLoS Genet 7: e1002225.

33. GloverDM (2012) The overlooked greatwall: a new perspective on mitotic control. Open Biol 2 : 120023.

34. LorcaT, CastroA (2012) The Greatwall kinase: a new pathway in the control of the cell cycle. Oncogene 32 : 537–543.

35. HarveySL, EncisoG, DephoureN, GygiSP, GunawardenaJ, et al. (2011) A phosphatase threshold sets the level of Cdk1 activity in early mitosis in budding yeast. Mol Biol Cell 22 : 3595–3608.

36. PalG, ParazMT, KelloggDR (2008) Regulation of Mih1/Cdc25 by protein phosphatase 2A and casein kinase 1. J Cell Biol 180 : 931–945.

37. LinFC, ArndtKT (1995) The role of Saccharomyces cerevisiae type 2A phosphatase in the actin cytoskeleton and in entry into mitosis. Embo J 14 : 2745–2759.

38. MinshullJ, StraightA, RudnerAD, DernburgAF, BelmontA, et al. (1996) Protein phosphatase 2A regulates MPF activity and sister chromatid cohesion in budding yeast. Curr Biol 6 : 1609–1620.

39. YangH, JiangW, GentryM, HallbergRL (2000) Loss of a protein phosphatase 2A regulatory subunit (Cdc55p) elicits improper regulation of Swe1p degradation. Mol Cell Biol 20 : 8143–8156.

40. TalarekN, CameroniE, JaquenoudM, LuoX, BontronS, et al. (2010) Initiation of the TORC1-regulated G0 program requires Igo1/2, which license specific mRNAs to evade degradation via the 5′-3′ mRNA decay pathway. Mol Cell 38 : 345–355.

41. BontronS, JaquenoudM, VagaS, TalarekN, BodenmillerB, et al. (2012) Yeast Endosulfines Control Entry into Quiescence and Chronological Life Span by Inhibiting Protein Phosphatase 2A. Cell Rep 3 : 16–22.

42. KimMY, BucciarelliE, MortonDG, WilliamsBC, Blake-HodekK, et al. (2012) Bypassing the Greatwall-Endosulfine Pathway: Plasticity of a Pivotal Cell-Cycle Regulatory Module in Drosophila melanogaster and Caenorhabditis elegans. Genetics 191 : 1181–1197.

43. RossioV, YoshidaS (2011) Spatial regulation of Cdc55-PP2A by Zds1/Zds2 controls mitotic entry and mitotic exit in budding yeast. J Cell Biol 193 : 445–454.

44. DulubovaI, HoriuchiA, SnyderGL, GiraultJA, CzernikAJ, et al. (2001) ARPP-16/ARPP-19: a highly conserved family of cAMP-regulated phosphoproteins. J Neurochem 77 : 229–238.

45. BreitkreutzA, ChoiH, SharomJR, BoucherL, NeduvaV, et al. (2010) A global protein kinase and phosphatase interaction network in yeast. Science 328 : 1043–1046.

46. WangH, WangX, JiangY (2003) Interaction with Tap42 is required for the essential function of Sit4 and type 2A phosphatases. Mol Biol Cell 14 : 4342–4351.

47. HombauerH, WeismannD, MudrakI, StanzelC, FellnerT, et al. (2007) Generation of active protein phosphatase 2A is coupled to holoenzyme assembly. PLoS Biol 5: e155.

48. ChiroliE, RossioV, LucchiniG, PiattiS (2007) The budding yeast PP2ACdc55 protein phosphatase prevents the onset of anaphase in response to morphogenetic defects. J Cell Biol 177 : 599–611.

49. QueraltE, UhlmannF (2008) Separase cooperates with Zds1 and Zds2 to activate Cdc14 phosphatase in early anaphase. J Cell Biol 182 : 873–883.

50. YasutisK, VignaliM, RyderM, TameireF, DigheSA, et al. (2010) Zds2p regulates Swe1p-dependent polarized cell growth in Saccharomyces cerevisiae via a novel Cdc55p interaction domain. Mol Biol Cell 21 : 4373–4386.

51. HealyAM, ZolnierowiczS, StapletonAE, GoeblM, DePaoli-RoachAA, et al. (1991) CDC55, a Saccharomyces cerevisiae gene involved in cellular morphogenesis: identification, characterization, and homology to the B subunit of mammalian type 2A protein phosphatase. Mol Cell Biol 11 : 5767–5780.

52. QueraltE, LehaneC, NovakB, UhlmannF (2006) Downregulation of PP2A(Cdc55) phosphatase by separase initiates mitotic exit in budding yeast. Cell 125 : 719–732.

53. WangY, BurkeDJ (1997) Cdc55p, the B-type regulatory subunit of protein phosphatase 2A, has multiple functions in mitosis and is required for the kinetochore/spindle checkpoint in Saccharomyces cerevisiae. Mol Cell Biol 17 : 620–626.

54. WangY, NgTY (2006) Phosphatase 2A negatively regulates mitotic exit in Saccharomyces cerevisiae. Mol Biol Cell 17 : 80–89.

55. YellmanCM, BurkeDJ (2006) The role of Cdc55 in the spindle checkpoint is through regulation of mitotic exit in Saccharomyces cerevisiae. Mol Biol Cell 17 : 658–666.

56. SanthanamA, HartleyA, DuvelK, BroachJR, GarrettS (2004) PP2A phosphatase activity is required for stress and Tor kinase regulation of yeast stress response factor Msn2p. Eukaryot Cell 3 : 1261–1271.

57. Di ComoCJ, ArndtKT (1996) Nutrients, via the Tor proteins, stimulate the association of Tap42 with type 2A phosphatases. Genes Dev 10 : 1904–1916.

58. JiangY, BroachJR (1999) Tor proteins and protein phosphatase 2A reciprocally regulate Tap42 in controlling cell growth in yeast. EMBO J 18 : 2782–2792.

59. GavinAC, AloyP, GrandiP, KrauseR, BoescheM, et al. (2006) Proteome survey reveals modularity of the yeast cell machinery. Nature 440 : 631–636.

60. HoY, GruhlerA, HeilbutA, BaderGD, MooreL, et al. (2002) Systematic identification of protein complexes in Saccharomyces cerevisiae by mass spectrometry. Nature 415 : 180–183.

61. Hood-DeGrenierJK (2011) Identification of phosphatase 2A-like Sit4-mediated signalling and ubiquitin-dependent protein sorting as modulators of caffeine sensitivity in S. cerevisiae. Yeast 28 : 189–204.

62. ReinkeA, ChenJC, AronovaS, PowersT (2006) Caffeine targets TOR complex I and provides evidence for a regulatory link between the FRB and kinase domains of Tor1p. J Biol Chem 281 : 31616–31626.

63. HoltLJ, TuchBB, VillenJ, JohnsonAD, GygiSP, et al. (2009) Global analysis of Cdk1 substrate phosphorylation sites provides insights into evolution. Science 325 : 1682–1686.

64. BurgessA, VigneronS, BrioudesE, LabbeJC, LorcaT, et al. (2010) Loss of human Greatwall results in G2 arrest and multiple mitotic defects due to deregulation of the cyclin B-Cdc2/PP2A balance. Proc Natl Acad Sci U S A 107 : 12564–12569.

65. WickyS, TjandraH, SchieltzD, YatesJ3rd, KelloggDR (2011) The Zds proteins control entry into mitosis and target protein phosphatase 2A to the Cdc25 phosphatase. Mol Biol Cell 22 : 20–32.

66. DuvelK, SanthanamA, GarrettS, SchneperL, BroachJR (2003) Multiple roles of Tap42 in mediating rapamycin-induced transcriptional changes in yeast. Mol Cell 11 : 1467–1478.

67. JacintoE, GuoB, ArndtKT, SchmelzleT, HallMN (2001) TIP41 interacts with TAP42 and negatively regulates the TOR signaling pathway. Mol Cell 8 : 1017–1026.

68. YanG, ShenX, JiangY (2006) Rapamycin activates Tap42-associated phosphatases by abrogating their association with Tor complex 1. EMBO J 25 : 3546–3555.

69. UhlmannF (2001) Secured cutting: controlling separase at the metaphase to anaphase transition. EMBO Rep 2 : 487–492.

70. RahalR, AmonA (2008) Mitotic CDKs control the metaphase-anaphase transition and trigger spindle elongation. Genes Dev 22 : 1534–1548.

71. LongtineMS, TheesfeldCL, McMillanJN, WeaverE, PringleJR, et al. (2000) Septin-dependent assembly of a cell cycle-regulatory module in Saccharomyces cerevisiae. Mol Cell Biol 20 : 4049–4061.

72. McMillanJN, LongtineMS, SiaRA, TheesfeldCL, BardesES, et al. (1999) The morphogenesis checkpoint in Saccharomyces cerevisiae: cell cycle control of Swe1p degradation by Hsl1p and Hsl7p. Mol Cell Biol 19 : 6929–6939.

73. Domingo-SananesMR, KapuyO, HuntT, NovakB (2011) Switches and latches: a biochemical tug-of-war between the kinases and phosphatases that control mitosis. Philos Trans R Soc Lond B Biol Sci 366 : 3584–3594.

74. Maniatis T, Fritsch EF, Sambrook J (1992) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor Laboratory, NY.

75. ShermanF (1991) Getting started with yeast. Methods Enzymol 194 : 3–21.

76. WachA, BrachatA, PohlmannR, PhilippsenP (1994) New heterologous modules for classical or PCR-based gene disruptions in Saccharomyces cerevisiae. Yeast 10 : 1793–1808.

77. 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.

78. SheffMA, ThornKS (2004) Optimized cassettes for fluorescent protein tagging in Saccharomyces cerevisiae. Yeast 21 : 661–670.

79. RancatiG, CrispoV, LucchiniG, PiattiS (2005) Mad3/BubR1 phosphorylation during spindle checkpoint activation depends on both Polo and Aurora kinases in budding yeast. Cell Cycle 4 : 972–980.

80. SchwobE, BohmT, MendenhallMD, NasmythK (1994) The B-type cyclin kinase inhibitor p40SIC1 controls the G1 to S transition in S. cerevisiae [see comments] [published erratum appears in Cell 1996 Jan 12;84(1):following 174]. Cell 79 : 233–244.

81. SuranaU, AmonA, DowzerC, McGrewJ, ByersB, et al. (1993) Destruction of the CDC28/CLB mitotic kinase is not required for the metaphase to anaphase transition in budding yeast. Embo J 12 : 1969–1978.

82. FraschiniR, D'AmbrosioC, VenturettiM, LucchiniG, PiattiS (2006) Disappearance of the budding yeast Bub2-Bfa1 complex from the mother-bound spindle pole contributes to mitotic exit. J Cell Biol 172 : 335–346.

83. FellnerT, LacknerDH, HombauerH, PiribauerP, MudrakI, et al. (2003) A novel and essential mechanism determining specificity and activity of protein phosphatase 2A (PP2A) in vivo. Genes Dev 17 : 2138–2150.

84. FraschiniR, FormentiE, LucchiniG, PiattiS (1999) Budding yeast Bub2 is localized at spindle pole bodies and activates the mitotic checkpoint via a different pathway from Mad2. J Cell Biol 145 : 979–991.

85. CalabriaI, BaroB, Rodriguez-RodriguezJA, RussinolN, QueraltE (2012) Zds1 regulates PP2ACdc55 activity and Cdc14 activation during mitotic exit through its Zds_C motif. J Cell Sci 125 : 2875–2884.