PP2A-Twins Is Antagonized by Greatwall and Collaborates with Polo for Cell Cycle Progression and Centrosome Attachment to Nuclei in Drosophila Embryos
Cell division and development are regulated by networks of kinases and phosphatases. In early Drosophila embryogenesis, 13 rapid nuclear divisions take place in a syncytium, requiring fine coordination between cell cycle regulators. The Polo kinase is a conserved, crucial regulator of M-phase. We have recently reported an antagonism between Polo and Greatwall (Gwl), another mitotic kinase, in Drosophila embryos. However, the nature of the pathways linking them remained elusive. We have conducted a comprehensive screen for additional genes functioning with polo and gwl. We uncovered a strong interdependence between Polo and Protein Phosphatase 2A (PP2A) with its B-type subunit Twins (Tws). Reducing the maternal contribution of Polo and PP2A-Tws together is embryonic lethal. We found that Polo and PP2A-Tws collaborate to ensure centrosome attachment to nuclei. While a reduction in Polo activity leads to centrosome detachments observable mostly around prophase, a reduction in PP2A-Tws activity leads to centrosome detachments at mitotic exit, and a reduction in both Polo and PP2A-Tws enhances the frequency of detachments at all stages. Moreover, we show that Gwl antagonizes PP2A-Tws function in both meiosis and mitosis. Our study highlights how proper coordination of mitotic entry and exit is required during embryonic cell cycles and defines important roles for Polo and the Gwl-PP2A-Tws pathway in this process.
Vyšlo v časopise:
PP2A-Twins Is Antagonized by Greatwall and Collaborates with Polo for Cell Cycle Progression and Centrosome Attachment to Nuclei in Drosophila Embryos. PLoS Genet 7(8): e32767. doi:10.1371/journal.pgen.1002227
Kategorie:
Research Article
prolekare.web.journal.doi_sk:
https://doi.org/10.1371/journal.pgen.1002227
Souhrn
Cell division and development are regulated by networks of kinases and phosphatases. In early Drosophila embryogenesis, 13 rapid nuclear divisions take place in a syncytium, requiring fine coordination between cell cycle regulators. The Polo kinase is a conserved, crucial regulator of M-phase. We have recently reported an antagonism between Polo and Greatwall (Gwl), another mitotic kinase, in Drosophila embryos. However, the nature of the pathways linking them remained elusive. We have conducted a comprehensive screen for additional genes functioning with polo and gwl. We uncovered a strong interdependence between Polo and Protein Phosphatase 2A (PP2A) with its B-type subunit Twins (Tws). Reducing the maternal contribution of Polo and PP2A-Tws together is embryonic lethal. We found that Polo and PP2A-Tws collaborate to ensure centrosome attachment to nuclei. While a reduction in Polo activity leads to centrosome detachments observable mostly around prophase, a reduction in PP2A-Tws activity leads to centrosome detachments at mitotic exit, and a reduction in both Polo and PP2A-Tws enhances the frequency of detachments at all stages. Moreover, we show that Gwl antagonizes PP2A-Tws function in both meiosis and mitosis. Our study highlights how proper coordination of mitotic entry and exit is required during embryonic cell cycles and defines important roles for Polo and the Gwl-PP2A-Tws pathway in this process.
Zdroje
1. MorganDO 2007 The Cell Cycle: Principles of Control. London New Science Press 297
2. ArchambaultVGloverDM 2009 Polo-like kinases: conservation and divergence in their functions and regulation. Nat Rev Mol Cell Biol 10 265 275
3. CarmenaMRuchaudSEarnshawWC 2009 Making the Auroras glow: regulation of Aurora A and B kinase function by interacting proteins. Curr Opin Cell Biol 21 796 805
4. BloomJCrossFR 2007 Multiple levels of cyclin specificity in cell-cycle control. Nat Rev Mol Cell Biol 8 149 160
5. Trinkle-MulcahyLLamondAI 2006 Mitotic phosphatases: no longer silent partners. Curr Opin Cell Biol 18 623 631
6. De WulfPMontaniFVisintinR 2009 Protein phosphatases take the mitotic stage. Curr Opin Cell Biol 21 806 815
7. StegmeierFAmonA 2004 Closing mitosis: the functions of the Cdc14 phosphatase and its regulation. Annu Rev Genet 38 203 232
8. SchmitzMHHeldMJanssensVHutchinsJRHudeczO 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
9. MochidaSIkeoSGannonJHuntT 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
10. FoeVEOdellGMEdgarBA 1993 Mitosis and morphogenesis in the Drosophila embryo: point and counterpoint. BateMAriasAMartinez The Development of Drosophila melanogaster New York Cold Spring Harbor Laboratory Press 746
11. BakerJTheurkaufWESchubigerG 1993 Dynamic changes in microtubule configuration correlate with nuclear migration in the preblastoderm Drosophila embryo. J Cell Biol 122 113 121
12. HiraokaYAgardDASedatJW 1990 Temporal and spatial coordination of chromosome movement, spindle formation, and nuclear envelope breakdown during prometaphase in Drosophila melanogaster embryos. J Cell Biol 111 2815 2828
13. StafstromJPStaehelinLA 1984 Dynamics of the nuclear envelope and of nuclear pore complexes during mitosis in the Drosophila embryo. Eur J Cell Biol 34 179 189
14. RothwellWFSullivanW 2000 The centrosome in early Drosophila embryogenesis. Curr Top Dev Biol 49 409 447
15. Rodrigues-MartinsARiparbelliMCallainiGGloverDMBettencourt-DiasM 2008 From centriole biogenesis to cellular function: centrioles are essential for cell division at critical developmental stages. Cell Cycle 7 11 16
16. BastoRLauJVinogradovaTGardiolAWoodsCG 2006 Flies without centrioles. Cell 125 1375 1386
17. SunkelCEGloverDM 1988 polo, a mitotic mutant of Drosophila displaying abnormal spindle poles. J Cell Sci 89 Pt1 25 38
18. PetronczkiMLenartPPetersJM 2008 Polo on the Rise-from Mitotic Entry to Cytokinesis with Plk1. Dev Cell 14 646 659
19. KumagaiADunphyWG 1996 Purification and molecular cloning of Plx1, a Cdc25-regulatory kinase from Xenopus egg extracts. Science 273 1377 1380
20. van VugtMAMedemaRH 2005 Getting in and out of mitosis with Polo-like kinase-1. Oncogene 24 2844 2859
21. MargalitAVlcekSGruenbaumYFoisnerR 2005 Breaking and making of the nuclear envelope. J Cell Biochem 95 454 465
22. LindqvistARodriguez-BravoVMedemaRH 2009 The decision to enter mitosis: feedback and redundancy in the mitotic entry network. J Cell Biol 185 193 202
23. ArchambaultVZhaoXWhite-CooperHCarpenterATGloverDM 2007 Mutations in Drosophila Greatwall/Scant Reveal Its Roles in Mitosis and Meiosis and Interdependence with Polo Kinase. PLoS Genet 3 e200 doi:10.1371/journal.pgen.0030200
24. YuJFlemingSLWilliamsBWilliamsEVLiZ 2004 Greatwall kinase: a nuclear protein required for proper chromosome condensation and mitotic progression in Drosophila. J Cell Biol 164 487 492
25. YuJZhaoYLiZGalasSGoldbergML 2006 Greatwall kinase participates in the Cdc2 autoregulatory loop in Xenopus egg extracts. Mol Cell 22 83 91
26. BurgessAVigneronSBrioudesELabbeJCLorcaT 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
27. VoetsEWolthuisRM 2010 MASTL is the human orthologue of Greatwall kinase that facilitates mitotic entry, anaphase and cytokinesis. Cell Cycle 9 3591 3601
28. ArchambaultVPinsonX 2010 Free centrosomes: where do they all come from? Fly (Austin) 4 172 177
29. SibonOCKelkarALemstraWTheurkaufWE 2000 DNA-replication/DNA-damage-dependent centrosome inactivation in Drosophila embryos. Nat Cell Biol 2 90 95
30. TakadaSKelkarATheurkaufWE 2003 Drosophila checkpoint kinase 2 couples centrosome function and spindle assembly to genomic integrity. Cell 113 87 99
31. ArchambaultVD'AvinoPPDeeryMJLilleyKSGloverDM 2008 Sequestration of Polo kinase to microtubules by phosphopriming-independent binding to Map205 is relieved by phosphorylation at a CDK site in mitosis. Genes Dev 22 2707 2720
32. SteegmaierMHoffmannMBaumALenartPPetronczkiM 2007 BI 2536, a potent and selective inhibitor of polo-like kinase 1, inhibits tumor growth in vivo. Curr Biol 17 316 322
33. BarrosTPKinoshitaKHymanAARaffJW 2005 Aurora A activates D-TACC-Msps complexes exclusively at centrosomes to stabilize centrosomal microtubules. J Cell Biol 170 1039 1046
34. LiHLiuXSYangXSongBWangY 2010 Polo-like kinase 1 phosphorylation of p150Glued facilitates nuclear envelope breakdown during prophase. Proc Natl Acad Sci U S A 107 14633 14638
35. ChaseDSerafinasCAshcroftNKosinskiMLongoD 2000 The polo-like kinase PLK-1 is required for nuclear envelope breakdown and the completion of meiosis in Caenorhabditis elegans. Genesis 26 26 41
36. RyderEAshburnerMBautista-LlacerRDrummondJWebsterJ 2007 The DrosDel deletion collection: a Drosophila genomewide chromosomal deficiency resource. Genetics 177 615 629
37. White-CooperHCarmenaMGonzalezCGloverDM 1996 Mutations in new cell cycle genes that fail to complement a multiply mutant third chromosome of Drosophila. Genetics 144 1097 1111
38. Mayer-JaekelREOhkuraHGomesRSunkelCEBaumgartnerS 1993 The 55 kd regulatory subunit of Drosophila protein phosphatase 2A is required for anaphase. Cell 72 621 633
39. ChenFArchambaultVKarALioPD'AvinoPP 2007 Multiple protein phosphatases are required for mitosis in Drosophila. Curr Biol 17 293 303
40. DeakPDonaldsonMGloverDM 2003 Mutations in makos, a Drosophila gene encoding the Cdc27 subunit of the anaphase promoting complex, enhance centrosomal defects in polo and are suppressed by mutations in twins/aar, which encodes a regulatory subunit of PP2A. J Cell Sci 116 4147 4158
41. JanssensVGorisJ 2001 Protein phosphatase 2A: a highly regulated family of serine/threonine phosphatases implicated in cell growth and signalling. Biochem J 353 417 439
42. SnaithHAArmstrongCGGuoYKaiserKCohenPT 1996 Deficiency of protein phosphatase 2A uncouples the nuclear and centrosome cycles and prevents attachment of microtubules to the kinetochore in Drosophila microtubule star (mts) embryos. J Cell Sci 109 Pt13 3001 3012
43. CastilhoPVWilliamsBCMochidaSZhaoYGoldbergML 2009 The M phase kinase Greatwall (Gwl) promotes inactivation of PP2A/B55delta, a phosphatase directed against CDK phosphosites. Mol Biol Cell 20 4777 4789
44. VigneronSBrioudesEBurgessALabbeJCLorcaT 2009 Greatwall maintains mitosis through regulation of PP2A. Embo J 28 2786 2793
45. KotadiaSKaoLRComerfordSAJonesRTHammerRE 2008 PP2A-dependent disruption of centrosome replication and cytoskeleton organization in Drosophila by SV40 small tumor antigen. Oncogene 27 6334 6346
46. ForesterCMMaddoxJLouisJVGorisJVirshupDM 2007 Control of mitotic exit by PP2A regulation of Cdc25C and Cdk1. Proc Natl Acad Sci U S A 104 19867 19872
47. WangCChangKCSomersGVirshupDAngBT 2009 Protein phosphatase 2A regulates self-renewal of Drosophila neural stem cells. Development 136 2287 2296
48. DobbelaereJJosueFSuijkerbuijkSBaumBTaponN 2008 A genome-wide RNAi screen to dissect centriole duplication and centrosome maturation in Drosophila. PLoS Biol 6 e224 doi:10.1371/journal.pbio.0060224
49. Mayer-JaekelREOhkuraHFerrignoPAndjelkovicNShiomiK 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 Pt9 2609 2616
50. BardinAJAmonA 2001 Men and sin: what's the difference? Nat Rev Mol Cell Biol 2 815 826
51. Gharbi-AyachiALabbeJCBurgessAVigneronSStrubJM 2010 The substrate of Greatwall kinase, Arpp19, controls mitosis by inhibiting protein phosphatase 2A. Science 330 1673 1677
52. MochidaSMaslenSLSkehelMHuntT 2010 Greatwall phosphorylates an inhibitor of protein phosphatase 2A that is essential for mitosis. Science 330 1670 1673
53. RangoneHWegelEGattMKYeungEFlowers A etal 2011 Suppression of Scant Identifies Endos as a Substrate of Greatwall Kinase and a Negative Regulator of Protein Phosphatase 2A in Mitosis. PLoS Gen 7 8 e1002225 doi:10.1371/journal.pgen.1002225
54. Von StetinaJRTranguchSDeySKLeeLAChaB 2008 alpha-Endosulfine is a conserved protein required for oocyte meiotic maturation in Drosophila. Development 135 3697 3706
Štítky
Genetika Reprodukčná medicínaČlánok vyšiel v časopise
PLOS Genetics
2011 Číslo 8
- Je „freeze-all“ pro všechny? Odborníci na fertilitu diskutovali na virtuálním summitu
- Gynekologové a odborníci na reprodukční medicínu se sejdou na prvním virtuálním summitu
Najčítanejšie v tomto čísle
- An EMT–Driven Alternative Splicing Program Occurs in Human Breast Cancer and Modulates Cellular Phenotype
- Chromosome Painting Reveals Asynaptic Full Alignment of Homologs and HIM-8–Dependent Remodeling of Chromosome Territories during Meiosis
- Discovery of Sexual Dimorphisms in Metabolic and Genetic Biomarkers
- Regulation of p53/CEP-1–Dependent Germ Cell Apoptosis by Ras/MAPK Signaling