Supplemental References

S10. Margolis, S.S., Walsh, S., Weiser, D.C., Yoshida, M., Shenolikar, S., and Kornbluth, S. (2003). PP1 control of M phase entry exerted through 14-3-3-regulated Cdc25 dephosphorylation. Embo J 22, 5734-5745. S11. Margolis, S.S., Perry, J.A., Weitzel, D.H., Freel, C.D., Yoshida, M., Haystead, T.A., and Kornbluth, S. (2006). A role for PP1 in the Cdc2/ B-mediated positive feedback activation of Cdc25. Mol Biol Cell 17, 1779-1789. S12. Axton, J.M., Dombradi, V., Cohen, P.T., and Glover, D.M. (1990). One of the protein 1 isoenzymes in Drosophila is essential for . Cell 63, 33-46. S13. Rogers, E., Bishop, J.D., Waddle, J.A., Schumacher, J.M., and Lin, R. (2002). The aurora AIR-2 functions in the release of chromosome cohesion in . J Cell Biol 157, 219-229. S14. Sassoon, I., Severin, F.F., Andrews, P.D., Taba, M.R., Kaplan, K.B., Ashford, A.J., Stark, M.J., Sorger, P.K., and Hyman, A.A. (1999). Regulation of Saccharomyces cerevisiae kinetochores by the type 1 phosphatase Glc7p. Genes Dev 13, 545-555. S15. Katayama, H., Zhou, H., Li, Q., Tatsuka, M., and Sen, S. (2001). Interaction and feedback regulation between STK15/BTAK/Aurora-A kinase and protein phosphatase 1 through mitotic cycle. J Biol Chem 276, 46219-46224. S16. Ohashi, S., Sakashita, G., Ban, R., Nagasawa, M., Matsuzaki, H., Murata, Y., Taniguchi, H., Shima, H., Furukawa, K., and Urano, T. (2006). Phospho-regulation of human protein kinase Aurora-A: analysis using anti-phospho-Thr288 monoclonal antibodies. 25, 7691-7702. S17. Helps, N.R., Luo, X., Barker, H.M., and Cohen, P.T. (2000). NIMA-related kinase 2 (Nek2), a cell-cycle-regulated protein kinase localized to centrosomes, is complexed to protein phosphatase 1. Biochem J 349, 509-518. S18. Glover, D.M., Leibowitz, M.H., McLean, D.A., and Parry, H. (1995). Mutations in aurora prevent centrosome separation leading to the formation of monopolar spindles. Cell 81, 95-105. S19. Meraldi, P., and Nigg, E.A. (2001). Centrosome cohesion is regulated by a balance of kinase and phosphatase activities. J Cell Sci 114, 3749-3757. S20. Eto, M., Elliott, E., Prickett, T.D., and Brautigan, D.L. (2002). Inhibitor-2 regulates protein phosphatase-1 complexed with NimA-related kinase to induce centrosome separation. J Biol Chem 277, 44013-44020. S21. Hsu, J.Y., Sun, Z.W., Li, X., Reuben, M., Tatchell, K., Bishop, D.K., Grushcow, J.M., Brame, C.J., Caldwell, J.A., Hunt, D.F., Lin, R., Smith, M.M., and Allis, C.D. (2000). Mitotic phosphorylation of histone H3 is governed by Ipl1/aurora kinase and Glc7/PP1 phosphatase in budding and . Cell 102, 279-291. S22. Sugiyama, K., Sugiura, K., Hara, T., Sugimoto, K., Shima, H., Honda, K., Furukawa, K., Yamashita, S., and Urano, T. (2002). Aurora-B associated protein as negative regulators of kinase activation. Oncogene 21, 3103- 3111. S23. Goto, H., Yasui, Y., Nigg, E.A., and Inagaki, M. (2002). Aurora-B phosphorylates Histone H3 at serine28 with regard to the mitotic chromosome condensation. Genes Cells 7, 11-17. S24. Dombradi, V., Axton, J.M., Barker, H.M., and Cohen, P.T. (1990). Protein phosphatase 1 activity in Drosophila mutants with abnormalities in mitosis and chromosome condensation. FEBS Lett 275, 39-43. S25. Dombradi, V., and Cohen, P.T. (1992). Protein phosphorylation is involved in the regulation of chromatin condensation during . FEBS Lett 312, 21-26. S26. Baksa, K., Morawietz, H., Dombradi, V., Axton, M., Taubert, H., Szabo, G., Torok, I., Udvardy, A., Gyurkovics, H., Szoor, B., and et al. (1993). Mutations in the protein phosphatase 1 gene at 87B can differentially affect suppression of position-effect variegation and mitosis in . Genetics 135, 117-125. S27. Bloecher, A., and Tatchell, K. (2000). Dynamic localization of protein phosphatase type 1 in the mitotic of Saccharomyces cerevisiae. J Cell Biol 149, 125-140. S28. Zeitlin, S.G., Shelby, R.D., and Sullivan, K.F. (2001). CENP-A is phosphorylated by Aurora B kinase and plays an unexpected role in completion of . J Cell Biol 155, 1147-1157. S29. Andrews, P.D., and Stark, M.J. (2000). Type 1 protein phosphatase is required for maintenance of cell wall integrity, morphogenesis and cell cycle progression in Saccharomyces cerevisiae. J Cell Sci 113 ( Pt 3), 507-520. S30. Cheng, A., Dean, N.M., and Honkanen, R.E. (2000). Serine/threonine protein phosphatase type 1gamma1 is required for the completion of cytokinesis in human A549 lung carcinoma cells. J Biol Chem 275, 1846-1854. S31. Vereshchagina, N., Bennett, D., Szoor, B., Kirchner, J., Gross, S., Vissi, E., White-Cooper, H., and Alphey, L. (2004). The essential role of PP1beta in Drosophila is to regulate nonmuscle myosin. Mol Biol Cell 15, 4395-4405. S32. Kinoshita, N., Yamano, H., Niwa, H., Yoshida, T., and Yanagida, M. (1993). Negative regulation of mitosis by the fission yeast protein phosphatase ppa2. Genes Dev 7, 1059-1071. S33. Janssens, V., and Goris, J. (2001). Protein phosphatase 2A: a highly regulated family of serine/threonine phosphatases implicated in and signalling. Biochem J 353, 417-439. S34. Lin, F.C., and Arndt, K.T. (1995). The role of Saccharomyces cerevisiae type 2A phosphatase in the actin cytoskeleton and in entry into mitosis. Embo J 14, 2745-2759. S35. Evans, D.R., and Stark, M.J. (1997). Mutations in the Saccharomyces cerevisiae type 2A protein phosphatase catalytic subunit reveal roles in cell wall integrity, actin cytoskeleton organization and mitosis. Genetics 145, 227- 241. S36. Snaith, H.A., Armstrong, C.G., Guo, Y., Kaiser, K., and Cohen, P.T. (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 ( Pt 13), 3001-3012. S37. Sonnichsen, B., Koski, L.B., Walsh, A., Marschall, P., Neumann, B., Brehm, M., Alleaume, A.M., Artelt, J., Bettencourt, P., Cassin, E., Hewitson, M., Holz, C., Khan, M., Lazik, S., Martin, C., Nitzsche, B., Ruer, M., Stamford, J., Winzi, M., Heinkel, R., Roder, M., Finell, J., Hantsch, H., Jones, S.J., Jones, M., Piano, F., Gunsalus, K.C., Oegema, K., Gonczy, P., Coulson, A., Hyman, A.A., and Echeverri, C.J. (2005). Full-genome RNAi profiling of early embryogenesis in Caenorhabditis elegans. Nature 434, 462-469. S38. Van Hoof, C., and Goris, J. (2003). Phosphatases in : to be or not to be, PP2A is in the heart of the question. Biochim Biophys Acta 1640, 97-104. S39. Kitajima, T.S., Sakuno, T., Ishiguro, K.I., Iemura, S.I., Natsume, T., Kawashima, S.A., and Watanabe, Y. (2006). Shugoshin collaborates with protein phosphatase 2A to protect cohesin. Nature 441, 46-52. S40. Tang, Z., Shu, H., Qi, W., Mahmood, N., Mumby, M.C., and Yu, H. (2006). PP2A Is Required for Centromeric Localization of Sgo1 and Proper Chromosome Segregation. Dev Cell 10, 575-585. S41. Riedel, C.G., Katis, V.L., Katou, Y., Mori, S., Itoh, T., Helmhart, W., Galova, M., Petronczki, M., Gregan, J., Cetin, B., Mudrak, I., Ogris, E., Mechtler, K., Pelletier, L., Buchholz, F., Shirahige, K., and Nasmyth, K. (2006). Protein phosphatase 2A protects centromeric sister chromatid cohesion during meiosis I. Nature 441, 53-61. S42. Le Goff, X., Buvelot, S., Salimova, E., Guerry, F., Schmidt, S., Cueille, N., Cano, E., and Simanis, V. (2001). The protein phosphatase 2A B'-regulatory subunit par1p is implicated in regulation of the S. pombe septation initiation network. FEBS Lett 508, 136-142. S43. Jiang, W., and Hallberg, R.L. (2001). Correct regulation of the septation initiation network in Schizosaccharomyces pombe requires the activities of par1 and par2. Genetics 158, 1413-1429. S44. Deak, P., Donaldson, M., and Glover, D.M. (2003). Mutations in makos, a Drosophila gene encoding the Cdc27 subunit of the 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. S45. Minshull, J., Straight, A., Rudner, A.D., Dernburg, A.F., Belmont, A., and Murray, A.W. (1996). Protein phosphatase 2A regulates MPF activity and sister chromatid cohesion in budding yeast. Curr Biol 6, 1609-1620. S46. Wang, Y., and Burke, D.J. (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. S47. Wang, Y., and Ng, T.Y. (2006). Phosphatase 2A negatively regulates mitotic exit in Saccharomyces cerevisiae. Mol Biol Cell 17, 80-89. S48. Yellman, C.M., and Burke, D.J. (2006). The role of Cdc55 in the spindle checkpoint is through regulation of mitotic exit in Saccharomyces cerevisiae. Mol Biol Cell 17, 658-666. S49. Mayer-Jaekel, R.E., Ohkura, H., Gomes, R., Sunkel, C.E., Baumgartner, S., Hemmings, B.A., and Glover, D.M. (1993). The 55 kd regulatory subunit of Drosophila protein phosphatase 2A is required for anaphase. Cell 72, 621-633. S50. Kao, G., Tuck, S., Baillie, D., and Sundaram, M.V. (2004). C. elegans SUR-6/PR55 cooperates with LET-92/protein phosphatase 2A and promotes Raf activity independently of inhibitory Akt phosphorylation sites. Development 131, 755-765. S51. Fraser, A.G., Kamath, R.S., Zipperlen, P., Martinez-Campos, M., Sohrmann, M., and Ahringer, J. (2000). Functional genomic analysis of C. elegans chromosome I by systematic RNA interference. Nature 408, 325-330. S52. Kinoshita, K., Nemoto, T., Nabeshima, K., Kondoh, H., Niwa, H., and Yanagida, M. (1996). The regulatory subunits of fission yeast protein phosphatase 2A (PP2A) affect cell morphogenesis, cell wall synthesis and cytokinesis. Genes Cells 1, 29-45. S53. Cohen, P.T., Philp, A., and Vazquez-Martin, C. (2005). Protein phosphatase 4--from obscurity to vital functions. FEBS Lett 579, 3278-3286. S54. Hastie, C.J., Carnegie, G.K., Morrice, N., and Cohen, P.T. (2000). A novel 50 kDa protein forms complexes with protein phosphatase 4 and is located at centrosomal microtubule organizing centres. Biochem J 347 Pt 3, 845-855. S55. Brewis, N.D., Street, A.J., Prescott, A.R., and Cohen, P.T. (1993). PPX, a novel protein serine/threonine phosphatase localized to centrosomes. Embo J 12, 987-996. S56. Sumiyoshi, E., Sugimoto, A., and Yamamoto, M. (2002). Protein phosphatase 4 is required for centrosome maturation in mitosis and sperm meiosis in C. elegans. J Cell Sci 115, 1403-1410. S57. Helps, N.R., Brewis, N.D., Lineruth, K., Davis, T., Kaiser, K., and Cohen, P.T. (1998). Protein phosphatase 4 is an essential enzyme required for organisation of microtubules at centrosomes in Drosophila embryos. J Cell Sci 111 ( Pt 10), 1331-1340. S58. Sutton, A., Immanuel, D., and Arndt, K.T. (1991). The SIT4 protein phosphatase functions in late G1 for progression into . Mol Cell Biol 11, 2133-2148. S59. Fernandez-Sarabia, M.J., Sutton, A., Zhong, T., and Arndt, K.T. (1992). SIT4 protein phosphatase is required for the normal accumulation of SWI4, CLN1, CLN2, and HCS26 RNAs during late G1. Genes Dev 6, 2417-2428. S60. Shimanuki, M., Kinoshita, N., Ohkura, H., Yoshida, T., Toda, T., and Yanagida, M. (1993). Isolation and characterization of the fission yeast protein phosphatase gene ppe1+ involved in cell shape control and mitosis. Mol Biol Cell 4, 303-313. S61. Bastians, H., and Ponstingl, H. (1996). The novel human protein serine/threonine phosphatase 6 is a functional homologue of budding yeast Sit4p and fission yeast ppe1, which are involved in cell cycle regulation. J Cell Sci 109 ( Pt 12), 2865-2874. S62. Goshima, G., Iwasaki, O., Obuse, C., and Yanagida, M. (2003). The role of Ppe1/PP6 phosphatase for equal chromosome segregation in fission yeast kinetochore. Embo J 22, 2752-2763. S63. Ollendorff, V., and Donoghue, D.J. (1997). The serine/threonine phosphatase PP5 interacts with CDC16 and CDC27, two tetratricopeptide repeat-containing subunits of the anaphase-promoting complex. J Biol Chem 272, 32011- 32018. S64. Zeke, T., Morrice, N., Vazquez-Martin, C., and Cohen, P.T. (2005). Human protein phosphatase 5 dissociates from heat-shock proteins and is proteolytically activated in response to arachidonic acid and the microtubule- depolymerizing drug nocodazole. Biochem J 385, 45-56. S65. Zuo, Z., Urban, G., Scammell, J.G., Dean, N.M., McLean, T.K., Aragon, I., and Honkanen, R.E. (1999). Ser/Thr protein phosphatase type 5 (PP5) is a negative regulator of glucocorticoid receptor-mediated growth arrest. Biochemistry 38, 8849-8857. S66. Zuo, Z., Dean, N.M., and Honkanen, R.E. (1998). Serine/threonine protein phosphatase type 5 acts upstream of to regulate the induction of (WAF1/Cip1) and mediate growth arrest. J Biol Chem 273, 12250-12258. S67. Ali, A., Zhang, J., Bao, S., Liu, I., Otterness, D., Dean, N.M., Abraham, R.T., and Wang, X.F. (2004). Requirement of protein phosphatase 5 in DNA-damage-induced ATM activation. Genes Dev 18, 249-254. S68. Zhang, J., Bao, S., Furumai, R., Kucera, K.S., Ali, A., Dean, N.M., and Wang, X.F. (2005). Protein phosphatase 5 is required for ATR-mediated checkpoint activation. Mol Cell Biol 25, 9910-9919. S69. Wechsler, T., Chen, B.P., Harper, R., Morotomi-Yano, K., Huang, B.C., Meek, K., Cleaver, J.E., Chen, D.J., and Wabl, M. (2004). DNA-PKcs function regulated specifically by protein phosphatase 5. Proc Natl Acad Sci U S A 101, 1247- 1252. S70. Baril, C., and Therrien, M. (2006). Alphabet, a Ser/Thr phosphatase of the protein phosphatase 2C family, negatively regulates RAS/MAPK signaling in Drosophila. Dev Biol 294, 232-245. S71. Takekawa, M., Maeda, T., and Saito, H. (1998). Protein phosphatase 2Calpha inhibits the human stress-responsive p38 and JNK MAPK pathways. Embo J 17, 4744-4752. S72. Ofek, P., Ben-Meir, D., Kariv-Inbal, Z., Oren, M., and Lavi, S. (2003). Cell cycle regulation and p53 activation by protein phosphatase 2C alpha. J Biol Chem 278, 14299-14305. S73. Herrmann, L., Dittmar, T., and Erdmann, K.S. (2003). The protein tyrosine phosphatase PTP-BL associates with the midbody and is involved in the regulation of cytokinesis. Mol Biol Cell 14, 230-240. S74. Flint, A.J., Gebbink, M.F., Franza, B.R., Jr., Hill, D.E., and Tonks, N.K. (1993). Multi-site phosphorylation of the protein tyrosine phosphatase, PTP1B: identification of cell cycle regulated and phorbol ester stimulated sites of phosphorylation. Embo J 12, 1937-1946. S75. Schievella, A.R., Paige, L.A., Johnson, K.A., Hill, D.E., and Erikson, R.L. (1993). Protein tyrosine phosphatase 1B undergoes mitosis-specific phosphorylation on serine. Cell Growth Differ 4, 239-246. S76. Bukczynska, P., Klingler-Hoffmann, M., Mitchelhill, K.I., Lam, M.H., Ciccomancini, M., Tonks, N.K., Sarcevic, B., Kemp, B.E., and Tiganis, T. (2004). The T-cell protein tyrosine phosphatase is phosphorylated on Ser-304 by cyclin- dependent protein in mitosis. Biochem J 380, 939-949. S77. Karim, F.D., and Rubin, G.M. (1999). PTP-ER, a novel tyrosine phosphatase, functions downstream of Ras1 to downregulate MAP kinase during Drosophila eye development. Mol Cell 3, 741-750. S78. Rintelen, F., Hafen, E., and Nairz, K. (2003). The Drosophila dual-specificity ERK phosphatase DMKP3 cooperates with the ERK tyrosine phosphatase PTP-ER. Development 130, 3479-3490. S79. Niwa, R., Nagata-Ohashi, K., Takeichi, M., Mizuno, K., and Uemura, T. (2002). Control of actin reorganization by Slingshot, a family of phosphatases that dephosphorylate ADF/cofilin. Cell 108, 233-246. S80. Rogers, S.L., Wiedemann, U., Stuurman, N., and Vale, R.D. (2003). Molecular requirements for actin-based lamella formation in Drosophila S2 cells. J Cell Biol 162, 1079-1088. S81. Kaji, N., Ohashi, K., Shuin, M., Niwa, R., Uemura, T., and Mizuno, K. (2003). Cell cycle-associated changes in Slingshot phosphatase activity and roles in cytokinesis in cells. J Biol Chem 278, 33450-33455. S82. Sugiura, R., Toda, T., Shuntoh, H., Yanagida, M., and Kuno, T. (1998). pmp1+, a suppressor of calcineurin deficiency, encodes a novel MAP kinase phosphatase in fission yeast. Embo J 17, 140-148. S83. Martin-Blanco, E., Gampel, A., Ring, J., Virdee, K., Kirov, N., Tolkovsky, A.M., and Martinez-Arias, A. (1998). puckered encodes a phosphatase that mediates a feedback loop regulating JNK activity during dorsal closure in Drosophila. Genes Dev 12, 557-570. S84. McEwen, D.G., and Peifer, M. (2005). Puckered, a Drosophila MAPK phosphatase, ensures cell viability by antagonizing JNK-induced apoptosis. Development 132, 3935-3946. S85. Jordan, K.C., Schaeffer, V., Fischer, K.A., Gray, E.E., and Ruohola-Baker, H. (2006). Notch signaling through tramtrack bypasses the mitosis promoting activity of the JNK pathway in the mitotic-to-endocycle transition of Drosophila follicle cells. BMC Dev Biol 6, 16. S86. MacCorkle-Chosnek, R.A., VanHooser, A., Goodrich, D.W., Brinkley, B.R., and Tan, T.H. (2001). Cell cycle regulation of c-Jun N-terminal kinase activity at the centrosomes. Biochem Biophys Res Commun 289, 173-180. S87. MacCorkle, R.A., and Tan, T.H. (2004). Inhibition of JNK2 disrupts anaphase and produces aneuploidy in mammalian cells. J Biol Chem 279, 40112-40121. S88. Maehama, T., Taylor, G.S., and Dixon, J.E. (2001). PTEN and : novel phosphoinositide phosphatases. Annu Rev Biochem 70, 247-279. S89. Boutros, R., Dozier, C., and Ducommun, B. (2006). The when and wheres of CDC25 phosphatases. Curr Opin Cell Biol 18, 185-191. S90. Myer, D.L., Bahassi el, M., and Stambrook, P.J. (2005). The Plk3-Cdc25 circuit. Oncogene 24, 299-305. S91. Donzelli, M., and Draetta, G.F. (2003). Regulating mammalian checkpoints through Cdc25 inactivation. EMBO Rep 4, 671-677. S92. Smits, V.A., and Medema, R.H. (2001). Checking out the G(2)/M transition. Biochim Biophys Acta 1519, 1-12. S93. O'Brien, K.P., Remm, M., and Sonnhammer, E.L. (2005). Inparanoid: a comprehensive database of eukaryotic orthologs. Nucleic Acids Res 33, D476-480. S94. Remm, M., Storm, C.E., and Sonnhammer, E.L. (2001). Automatic clustering of orthologs and in-paralogs from pairwise species comparisons. J Mol Biol 314, 1041-1052.

Table S3. Drosophila Protein Phosphatases Required for Cell-Cycle Progression and Their Putative Orthologues

Gene Putative Gene Family Phenotypes in Current Study Previous Known Functions Name Orthologues Pp1-87B (HS) PPP1CA, PP1 catalytic Downregulation leads to an G2-M transition: required for M phase entry through 14-3-3- PPP1CC; (CE) increase in mitotic index with regulated Cdc25 dephosphorylation in Xenopus [S10, S11]. gsp-2; (SC) cells accumulating at Spindle assembly and chromosome stability: required for GLC7; (SP) , cell death and mitotic spindle organization and sister chromatid segregation in dis2 cells containing abnormally D. melanogaster [S12]; along with gsp-1, believed to play a role elongated anaphase-like in maintaining chromosome cohesion protein REC-8 to sustain spindles with unseparated chromosome cohesion by antagonizing the aurora kinase AIR-2 sister chromatids moving activity in C. elegans meiosis [S13]; required for kinetochore- toward to the same spindle microtubule interactions in S. cerevisiae [S14]. pole. An active spindle- Centrosome separation: inactives Aurora-A [S15, S16] and Nek2 assembly checkpoint and [S17], both of which are required for centrosome separation in D. undegraded in such melanogaster [S18] and in H. sapiens [S19, S20]. cells suggest a failure in Dephosphorylation of mitotic histone H3: antagonizes Aurora chromosome congression. kinase to dephosphorylate histone H3 in S. cerevisiae, C. Required for elegans [S21] and H. sapiens [S22, S23]; in addition, it is also dephosphorylating P-H3 upon implicated in regulation of chromosome condensation at mitotic exit. interphase in D. melanogaster [S24-S26]. Cytokinesis: present at cleavage furrow at the end of the M phase in S. cerevisiae [S27] and H. sapiens [S28]; conditional mutation in S. cerevisiae [S29] or microinjection of antisense oligonucleotides in H. sapiens [S30] resulted in defects in cytokinesis. flw (flap (HS)PPP1CB; PP1 catalytic RNAi cells or mutant larval This PP1 isoform previously shown to dephosphorylate myosin wing) (CE)gsp-1 neuroblasts display light chain in D. melanogaster [S31]. chromosome alignment/segregation defects. mts (HS)PPP2CB, PP2A Downregulation of either the G2-M transition: genetic, drug, and biochemical experiments (microtub PPP2CA; catalytic catalytic or structural A from S. pombe, Xenopus, and starfish indicate that PP2A ule star) (CE)let-92; subunit of PP2A leads to an negatively regulates the G2-M transition by interacting with the (SC)PPH22, increase in the mitotic index machinery involved in Cdc2 activation ([S32] and reviewed in PPH21; with cells accumulating at [S33]); evidence from S. cerevisiae however, supports a positive (SP)ppa2 prometaphase, abnormal role for PP2A in the G2-M transition [S34, S35]. PP2A- (HS)PPP2R1A PP2A chromosome behavior, and Spindle assembly: D. melanogaster mutants [S36] and C. 29B , PPP2R1B; regulatory aberrant elongated arrays of elegans RNAi embryos [S37] display defects in spindle (CE)paa-1; spindle microtubules assembly. (SC)TPD3; emanating from a single Apoptosis: inhibits apoptosis in D. melanogaster (reviewed in (SP)paa1 centrosomal mass. [S38]). Consistently, knockdown of the structural A subunit leads to instability of the catalytic subunit. Required for dephosphorylating P-H3 upon mitotic exit. wdb (HS)PPP2R5E, PP2A Downregulation leads to an Chromosome stability: associates with hSgo1 and prevents (widerbor PPP2R5A; regulatory increase in the mitotic index precocious cohesin dissociation at centromeres [S39, S40]; st) (CE)W08G11. with cells accumulating at associates with SP-Sgo1 and protect cohesin Rec8 from 4; (SP)Par1 prometaphase, defects in separase cleavage during meiosis I [S39, S41]. spindle assembly and Cytokinesis: negatively regulates S. pombe SIN signaling [S42, chromosome S43]. alignment/segregation; complexes with MEI-S332 and is required for the normal localization of MEI-S332 to kinetochores. tws/aar (HS)PPP2R2A PP2A Downregulation leads to a high -anaphase transition and chromosome stability: (twins/ , PPP2R2B, regulatory proportion of anaphase figures negatively regulates anaphase entry in D. melanogaster [S44] and abnormal PPP2R2D, showing lagging S. cerevisiae [S45-S48] through a mechanism independent of the anaphase PPP2R2C; chromosomes. known spindle checkpoint pathways; mutants from D. resolutio (CE)sur-6; melanogaster [S49] and C. elegans [S37, S50] display lagging n) (SC)CDC55; chromosomes and chromosome bridges. (SP)pab1 Spindle assembly: mutants from C. elegans display defective spindle assembly [S37, S50]. Cytokinesis: downregulation in C. elegans led to failure in cytokinesis [S37] [S51]; overexpression in S. pombe blocked septation, leading to an accumulation of binucleated cells [S52]. Pp4-19C (HS)PPP4C; PP4 catalytic Downregulation of either the Centrosome maturation: PP4 catalytic subunit localizes to (CE)pph-4.1; (PP2A-like) catalytic or regulatory subunit centrosomes during mitosis in D. melanogaster and C. elegans (SP)SPBC26H leads to an accumulation of G2 and to the pericentriolar material in human cells [S53]; human 8.05c cells associated with increased PPP4R2 binds to and colocalizes with PPP4C at centrosomal

PPP4R2r (HS)PPP4R2 PP4 cell size. microtubule organizing centers [S54, S55]; implicated in the regulatory recruitment of pericentriolar material (PCM) components to centrosomes in C. elegans and D. melanogaster [S56, S57]; Drosophila mutant exhibit mitotic spindle microtubules are either absent, or aberrant and unconnected to the centrosomes in syncytial embryos [S57]. PPV (HS)PPP6C; PP6 (PP2A- Downregulation leads to an Cell-cycle progression: required in late G1 for progression into (SC)SIT4; like) increase in mitotic index with S phase in S. cerevisiae [S58, S59]; mutants of S. pombe however, (SP)ppe1 cells accumulating at display G2 arrest [S60]; their human counterparts can prometaphase. functionally complement loss-of-function mutations of S. pombe ppe1 and S. cerevisiae SIT4 [S61]. Chromosome stability: bound to chromatin and mutants display unequal segregation of chromosomes under the challenge of microtubule inhibitor in S. pombe [S62] PpD3 (HS)PPP5C; PP5 Downregulation leads to an Cell-cycle control: in H. sapiens, the potential cell-cycle (CE)Y39B6A. increased frequency of both function of this gene has been implied by its capability to binds 2; mitotic cells with low and high to several cell-cycle regulators such as cdc16, cdc27 [S63], hsp70 (SC)SPBC3F6. numbers of centrosomes, and hsp90 [S64]; also implicated in a p53-dependent cell-cycle 01c; (SP)PPT1 perhaps indicating a role in arrest at G1 in response to glucocorticoids [S65, S66], S phase centrosome separation. cell-cycle arrest in response to ionizing irradiation and UV irradiation [S67, S68] and in DNA-repair processes after ionizing radiation [S69]. CG1906/ (HS)PPM1B, PP2C Downregulation leads to a Cell-cycle signaling: Drosophila CG1906/Alphabet negatively Alphabet PPM1A decrease in cells and regulates RAS/MAPK signaling [S70]; implicated in human p38 an increase in cells and JNK signaling pathway [S71] and p53-dependent G2-M associated with increased cell arrest and apoptosis [S72]. size. CG9311 (HS)PTPN23/ PTP Downregulation leads to Centrosome and cytokinesis: its human counterpart localizes PTP-BL; defective spindles and mis- at the centrosomes, the spindle midzone and the midbody, and (SC)BRO1; positioned centrosomes plays a regulatory role in cytokinesis [S73]. (SP)BRO1 detached from the spindle pole. domain protein csw (HS)PTPN11/ PTP RNAi cells or mutant larval N/A (corkscre PTP2C/SHP-2, neuroblasts display low mitotic w) PTPN6/PTP- indices and abnormal 1C/SHP-1; chromosome behaviour in (CE)ptp-2 mitosis. Ptp61F (HS)PTPN1/P PTP Downregulation leads to The potential cell-cycle functions of its human counterparts TP1B, abnormal chromosome have been suggested by the notion that both PTPN1 and PTPN2 PTPN2/TC- behaviour in mitosis, increased undergo mitosis-specific phosphorylation [S74-S76]. This PTP G2 phase cells, and increased phosphorylation may be mediated by CDKs (cyclin-dependent intermediate phase cells protein kinases) [S76]. between 2C and 4C. PTP-ER N/A PTP Downregulation leads to a Cell-cycle signaling: functions downstream of Ras1 to decrease in G1 phase cells, an downregulate ERK/MAPK [S77]; PTP-ER and DMKP3 perform increase in G2 phase cells and partially redundant functions on the same substrate, ERK [S78] abnormal chromosome behaviour in mitosis. ssh (HS)SSH3, DSP Downregulation leads to an Cytokinesis: regulates actin reorganization as downregulation (slingshot SSH1, SSH2; increase in mitotic index. of Ssh in D. melanogaster [S79, S80] or expression of a ) (SP)Pmp1 phosphatase-inactive SSH1 in H. sapiens [S81] led to abnormal accumulations of f-actin. Cytokinesis defects were not observed in Drosophila S2 cells treated with dsRNA to inhibit Ssh [S80]. In H. sapiens however, the phosphatase activity of SSH1 decreases in the early stages of mitosis and is elevated in and cytokinesis. It colocalizes with f-actin and accumulates onto the cleavage furrow and the midbody. Expression of a phosphatase-inactive SSH1 induces aberrant accumulation of f-actin and phospho-cofilin near the midbody in the final stage of cytokinesis and frequently leads to the regression of the cleavage furrow and the formation of multinucleate cells [S81]. Defective cytokinesis was also found in Pmp1 null mutant in S. pombe [S82]. puc (SC)MSG5, DSP Downregulation leads to a Cell-cycle signaling: the single phosphatase known to (puckered SDP1/YIL113 decrease in G1 phase cells, an antagonize Jun N-terminal kinase (JNK) signaling in D. ) W increase in G2 phase cells, melanogaster [S83, S84]. Lack of JNK activity results in the increased cell size, reduced initiation of premature endocycles and loss of Puckered leads to copies of centrosomes and loss of endocycles in the Drosophila follicle cells [S85]. defects in chromosome Centrosome functions: human JNK is reported to localize to alignment/segregation during centrosomes where it is active from early S through late mitosis. anaphase peaking at metaphase [S86, S87]. myotubul (HS)MTMR2, DSP Downregulation leads to N/A arin MTMR1, defects in chromosome MTM1; alignment/segregation during (CE)mtm-1 mitosis, a decrease in G1 phase cells, an increase in G2 phase cells and a significant increase in intermediate phase cells that may reflect aneuploidy arising from chromosome instability in mitosis. sbf (SET (HS)CMT4B2/ myotubularin- Downregulation leads to an N/A domain SBF2/MTMR1 like; anti- increase in mitotic index, binding 3, SBF1; phosphatase chromosome factor) (CE)mtm-5 alignment/segregation defects during mitosis, a decrease in G1 phase cells, an increase in G2 phase cells and a significant increase in intermediate phase cells that may reflect aneuploidy arising from chromosome instability in mitosis. CG3530 (HS)MTMR6, DSP; Downregulation leads to a N/A MTMR8; myotubularin- decrease in G1 phase cells and (CE)mtm-6; like an increase in G2 phase cells. (SC)YMR1; (SP)SPAC19A 8.03 Pten (HS)PTEN, DSP; PIP3 Downregulation leads to a Cell-cycle signaling: inhibits the activation of Akt (reviewed in PTENP1; phosphatase decrease in G1 phase cells and [S88]). (CE)F46F11.3; an increase in G2 phase cells (SP) ptn1 associated with increased cell size. string (HS)CDC25B, DSP Downregulation leads to an G2-M and G1-S transition: reviewed in [S89-S92]. (stg) CDC25C, accumulation of large G2 cells. CDC25A; (CE)R05H5.2; (SC)MIH1; (SP)cdc25

Abbreviations are as follows: HS, H. Sapiens; CE, C. elegans; SC, S. Cerevisiae; SP, S. Pombe. Orthologues of Drosophila protein phosphatases were obtained from other organisms in the Inparanoid eukaryotic database [S93] [S94] (http://inparanoid.cgb.ki.se/ confidence value = 0.05 or higher).