Journal of Cell Science, Vol 110, Issue 15 1793-1804, Copyright © 1997 by Company of Biologists

JOURNAL ARTICLES

Distinct subunit functions and cell cycle regulated phosphorylation of 20S APC/cyclosome required for anaphase in fission yeast

H Yamada, K Kumada and M Yanagida
Department of Biophysics, Graduate School of Science, Kyoto University, Japan.

We show here that the fission yeast gene products Cut9 and Nuc2 are the subunits of the 20S complex, the putative APC (anaphase promoting complex)/cyclosome which contains ubiquitin ligase activity required for cyclin and Cut2 destruction. The assembly of Cut9 into the 20S complex requires functional Nuc2, and vice versa. The size of fission yeast APC/cyclosome is similar to that of higher eukaryotes, but differs greatly from that (36S) of budding yeast. The 20S complex is present in cells arrested at different stages of the cell cycle, and becomes slightly heavier in mitosis than interphase. Cut9 in the 20S complex is hyperphosphorylated specifically at the time of metaphase. The truncated forms of Cut9 block entry into mitosis, however. The 20S assembly impaired in the cut9 mutant can be restored by elevating the level of a novel gene product Hcnl, similar to budding yeast Cdc26. Furthermore, deletion of protein kinase PKA (Pkal) suppresses the phenotype of the cut9 mutation and reduces phosphorylation of Cut9. In contrast, PP1 (Dis2) phosphatase mutation shows the reverse effect on the phenotype of cut9. The Cut9 subunit is likely to be a target for regulating APC/ cyclosome function through protein-protein interactions and phosphorylation.

This article has been cited by other articles:

				K. Narbonne-Reveau, S. Senger, M. Pal, A. Herr, H. E. Richardson, M. Asano, P. Deak, and M. A. Lilly APC/CFzr/Cdh1 promotes cell cycle progression during the Drosophila endocycle Development, April 15, 2008; 135(8): 1451 - 1461. [Abstract] [Full Text] [PDF]

				Y. Kimata, A. Matsuyama, K. Nagao, K. Furuya, C. Obuse, M. Yoshida, and M. Yanagida Diminishing HDACs by drugs or mutations promotes normal or abnormal sister chromatid separation by affecting APC/C and adherin J. Cell Sci., April 1, 2008; 121(7): 1107 - 1118. [Abstract] [Full Text] [PDF]

				J.-Y. Huang, G. Morley, D. Li, and M. Whitaker Cdk1 phosphorylation sites on Cdc27 are required for correct chromosomal localisation and APC/C function in syncytial Drosophila embryos J. Cell Sci., June 15, 2007; 120(12): 1990 - 1997. [Abstract] [Full Text] [PDF]

				Y. Kobayashi, S. Saitoh, Y. Ogiyama, S. Soejima, and K. Takahashi The fission yeast DASH complex is essential for satisfying the spindle assembly checkpoint induced by defects in the inner-kinetochore proteins Genes Cells, March 1, 2007; 12(3): 311 - 328. [Abstract] [Full Text] [PDF]

				S. A. Kawashima, T. Tsukahara, M. Langegger, S. Hauf, T. S. Kitajima, and Y. Watanabe Shugoshin enables tension-generating attachment of kinetochores by loading Aurora to centromeres Genes & Dev., February 15, 2007; 21(4): 420 - 435. [Abstract] [Full Text] [PDF]

				H.-J. Yoon, A. Feoktistova, J.-S. Chen, J. L. Jennings, A. J. Link, and K. L. Gould Role of Hcn1 and Its Phosphorylation in Fission Yeast Anaphase-promoting Complex/Cyclosome Function J. Biol. Chem., October 27, 2006; 281(43): 32284 - 32293. [Abstract] [Full Text] [PDF]

				J. U. Gutterman, H. T. Lai, P. Yang, V. Haridas, A. Gaikwad, and S. Marcus Effects of the tumor inhibitory triterpenoid avicin G on cell integrity, cytokinesis, and protein ubiquitination in fission yeast PNAS, September 6, 2005; 102(36): 12771 - 12776. [Abstract] [Full Text] [PDF]

				S. Saitoh, K. Ishii, Y. Kobayashi, and K. Takahashi Spindle Checkpoint Signaling Requires the Mis6 Kinetochore Subcomplex, Which Interacts with Mad2 and Mitotic Spindles Mol. Biol. Cell, August 1, 2005; 16(8): 3666 - 3677. [Abstract] [Full Text] [PDF]

				B. B. Gadea and J. V. Ruderman Aurora Kinase Inhibitor ZM447439 Blocks Chromosome-induced Spindle Assembly, the Completion of Chromosome Condensation, and the Establishment of the Spindle Integrity Checkpoint in Xenopus Egg Extracts Mol. Biol. Cell, March 1, 2005; 16(3): 1305 - 1318. [Abstract] [Full Text] [PDF]

				E. B. Gomez, V. T. Angeles, and S. L. Forsburg A Screen for Schizosaccharomyces pombe Mutants Defective in Rereplication Identifies New Alleles of rad4+, cut9+ and psf2+ Genetics, January 1, 2005; 169(1): 77 - 89. [Abstract] [Full Text] [PDF]

				A. Kerres, C. Vietmeier-Decker, J. Ortiz, I. Karig, C. Beuter, J. Hegemann, J. Lechner, and U. Fleig The Fission Yeast Kinetochore Component Spc7 Associates with the EB1 Family Member Mal3 and Is Required for Kinetochore-Spindle Association Mol. Biol. Cell, December 1, 2004; 15(12): 5255 - 5267. [Abstract] [Full Text] [PDF]

				Y. Kimata and M. Yanagida Suppression of a mitotic mutant by tRNA-Ala anticodon mutations that produce a dominant defect in late mitosis J. Cell Sci., May 1, 2004; 117(11): 2283 - 2293. [Abstract] [Full Text] [PDF]

				M. Schwickart, J. Havlis, B. Habermann, A. Bogdanova, A. Camasses, T. Oelschlaegel, A. Shevchenko, and W. Zachariae Swm1/Apc13 Is an Evolutionarily Conserved Subunit of the Anaphase-Promoting Complex Stabilizing the Association of Cdc16 and Cdc27 Mol. Cell. Biol., April 15, 2004; 24(8): 3562 - 3576. [Abstract] [Full Text] [PDF]

				R. de Lahondes, V. Ribes, and B. Arcangioli Fission Yeast Sap1 Protein Is Essential for Chromosome Stability Eukaryot. Cell, October 1, 2003; 2(5): 910 - 921. [Abstract] [Full Text] [PDF]

				M. Bolte, P. Dieckhoff, C. Krause, G. H. Braus, and S. Irniger Synergistic inhibition of APC/C by glucose and activated Ras proteins can be mediated by each of the Tpk1-3 proteins in Saccharomyces cerevisiae Microbiology, May 1, 2003; 149(5): 1205 - 1216. [Abstract] [Full Text] [PDF]

				Q.-W. Jin, A. L. Pidoux, C. Decker, R. C. Allshire, and U. Fleig The Mal2p Protein Is an Essential Component of the Fission Yeast Centromere Mol. Cell. Biol., October 15, 2002; 22(20): 7168 - 7183. [Abstract] [Full Text] [PDF]

				J. W. Harper, J. L. Burton, and M. J. Solomon The anaphase-promoting complex: it's not just for mitosis any more Genes & Dev., September 1, 2002; 16(17): 2179 - 2206. [Full Text] [PDF]

				A. E. Ikui, K. Furuya, M. Yanagida, and T. Matsumoto Control of localization of a spindle checkpoint protein, Mad2, in fission yeast J. Cell Sci., April 15, 2002; 115(8): 1603 - 1610. [Abstract] [Full Text] [PDF]

				M. H. Glickman and A. Ciechanover The Ubiquitin-Proteasome Proteolytic Pathway: Destruction for the Sake of Construction Physiol Rev, April 1, 2002; 82(2): 373 - 428. [Abstract] [Full Text] [PDF]

				M. J. Heitz, J. Petersen, S. Valovin, and I. M. Hagan MTOC formation during mitotic exit in fission yeast J. Cell Sci., March 14, 2002; 114(24): 4521 - 4532. [Abstract] [Full Text] [PDF]

				R. R. West, T. Malmstrom, and J. R. McIntosh Kinesins klp5+ and klp6+ are required for normal chromosome movement in mitosis J. Cell Sci., January 3, 2002; 115(5): 931 - 940. [Abstract] [Full Text] [PDF]

				A. M. Bentley, B. C. Williams, M. L. Goldberg, and A. J. Andres Phenotypic characterization of Drosophila ida mutants: defining the role of APC5 in cell cycle progression J. Cell Sci., January 3, 2002; 115(5): 949 - 961. [Abstract] [Full Text] [PDF]

				L. Chang, J. L. Morrell, A. Feoktistova, and K. L. Gould Study of Cyclin Proteolysis in Anaphase-Promoting Complex (APC) Mutant Cells Reveals the Requirement for APC Function in the Final Steps of the Fission Yeast Septation Initiation Network Mol. Cell. Biol., October 1, 2001; 21(19): 6681 - 6694. [Abstract] [Full Text] [PDF]

				M. Gmachl, C. Gieffers, A. V. Podtelejnikov, M. Mann, and J.-M. Peters The RING-H2 finger protein APC11 and the E2 enzyme UBC4 are sufficient to ubiquitinate substrates of the anaphase-promoting complex PNAS, August 1, 2000; 97(16): 8973 - 8978. [Abstract] [Full Text] [PDF]

				A. D. Rudner and A. W. Murray Phosphorylation by Cdc28 Activates the Cdc20-dependent Activity of the Anaphase-promoting Complex J. Cell Biol., June 26, 2000; 149(7): 1377 - 1390. [Abstract] [Full Text] [PDF]

				E. R. Kramer, N. Scheuringer, A. V. Podtelejnikov, M. Mann, and J.-M. Peters Mitotic Regulation of the APC Activator Proteins CDC20 and CDH1 Mol. Biol. Cell, May 1, 2000; 11(5): 1555 - 1569. [Abstract] [Full Text]

				S. Irniger, M. Bäumer, and G. H. Braus Glucose and Ras Activity Influence the Ubiquitin Ligases APC/C and SCF in Saccharomyces cerevisiae Genetics, April 1, 2000; 154(4): 1509 - 1521. [Abstract] [Full Text]

				H. Yamada, S Matsumoto, and T Matsumoto High dosage expression of a zinc finger protein, Grt1, suppresses a mutant of fission yeast slp1(+), a homolog of CDC20/p55CDC/Fizzy J. Cell Sci., January 11, 2000; 113(22): 3989 - 3999. [Abstract] [PDF]

				S. Kotani, H. Tanaka, H. Yasuda, and K. Todokoro Regulation of APC Activity by Phosphorylation and Regulatory Factors J. Cell Biol., August 23, 1999; 146(4): 791 - 800. [Abstract] [Full Text] [PDF]

				W. Zachariae and K. Nasmyth Whose end is destruction: cell division and the anaphase-promoting complex Genes & Dev., August 15, 1999; 13(16): 2039 - 2058. [Full Text]

				G. Goshima, S. Saitoh, and M. Yanagida Proper metaphase spindle length is determined by centromere proteins Mis12 and Mis6 required for faithful chromosome segregation Genes & Dev., July 1, 1999; 13(13): 1664 - 1677. [Abstract] [Full Text]

				D. C. Farruggio, F. M. Townsley, and J. V. Ruderman Cdc20 associates with the kinase aurora2/Aik PNAS, June 22, 1999; 96(13): 7306 - 7311. [Abstract] [Full Text] [PDF]

				R. Fraschini, E. Formenti, G. Lucchini, and S. Piatti Budding Yeast Bub2 Is Localized at Spindle Pole Bodies and Activates the Mitotic Checkpoint via a Different Pathway from Mad2 J. Cell Biol., May 31, 1999; 145(5): 979 - 991. [Abstract] [Full Text] [PDF]

				R. Grossberger, C. Gieffers, W. Zachariae, A. V. Podtelejnikov, A. Schleiffer, K. Nasmyth, M. Mann, and J.-M. Peters Characterization of the DOC1/APC10 Subunit of the Yeast and the Human Anaphase-promoting Complex J. Biol. Chem., May 14, 1999; 274(20): 14500 - 14507. [Abstract] [Full Text] [PDF]

				L. D. Berry, A. Feoktistova, M. D. Wright, and K. L. Gould The Schizosaccharomyces pombe dim1+ Gene Interacts with the Anaphase-Promoting Complex or Cyclosome (APC/C) Component lid1+ and Is Required for APC/C Function Mol. Cell. Biol., April 1, 1999; 19(4): 2535 - 2546. [Abstract] [Full Text] [PDF]

				I. Sassoon, F. F. Severin, P. D. Andrews, M.-R. Taba, K. B. Kaplan, A. J. Ashford, M. J.R. Stark, P. K. Sorger, and A. A. Hyman Regulation of Saccharomyces cerevisiae kinetochores by the type 1 phosphatase Glc7p Genes & Dev., March 1, 1999; 13(5): 545 - 555. [Abstract] [Full Text]

				K. Furuya, K. Takahashi, and M. Yanagida Faithful anaphase is ensured by Mis4, a sister chromatid cohesion molecule required in S phase and not destroyed in G1 phase Genes & Dev., November 1, 1998; 12(21): 3408 - 3418. [Abstract] [Full Text]

				R. R. West, E. V. Vaisberg, R. Ding, P. Nurse, and J. R. McIntosh cut11+: A Gene Required for Cell Cycle-dependent Spindle Pole Body Anchoring in the Nuclear Envelope and Bipolar Spindle Formation in Schizosaccharomyces pombe Mol. Biol. Cell, October 1, 1998; 9(10): 2839 - 2855. [Abstract] [Full Text]

				D. Patra and W. G. Dunphy Xe-p9, a Xenopus Suc1/Cks protein, is essential for the Cdc2-dependent phosphorylation of the anaphase- promoting complex at mitosis Genes & Dev., August 15, 1998; 12(16): 2549 - 2559. [Abstract] [Full Text]

				N. Rhind and P. Russell Tyrosine Phosphorylation of Cdc2 Is Required for the Replication Checkpoint in Schizosaccharomyces pombe Mol. Cell. Biol., July 1, 1998; 18(7): 3782 - 3787. [Abstract] [Full Text]

				E. L. Grishchuk, J. L. Howe, and J. R. McIntosh A Screen for Genes Involved in the Anaphase Proteolytic Pathway Identifies tsm1+, a Novel Schizosaccharomyces pombe Gene Important for Microtubule Integrity Genetics, July 1, 1998; 149(3): 1251 - 1264. [Abstract] [Full Text] [PDF]

				G. J. Gorbsky, R.-H. Chen, and A. W. Murray Microinjection of Antibody to Mad2 Protein into Mammalian Cells in Mitosis Induces Premature Anaphase J. Cell Biol., June 1, 1998; 141(5): 1193 - 1205. [Abstract] [Full Text] [PDF]

				C. Lies, J Cheng, S. James, N. Morris, M. O'Connell, and P. Mirabito BIMAAPC3, a component of the Aspergillus anaphase promoting complex/cyclosome, is required for a G2 checkpoint blocking entry into mitosis in the absence of NIMA function J. Cell Sci., January 5, 1998; 111(10): 1453 - 1465. [Abstract] [PDF]

				M. Sohrmann, S. Schmidt, I. Hagan, and V. Simanis Asymmetric segregation on spindle poles of the Schizosaccharomyces pombe septum-inducing protein kinase Cdc7p Genes & Dev., January 1, 1998; 12(1): 84 - 94. [Abstract] [Full Text]

				Y. Saka, F. Esashi, T. Matsusaka, S. Mochida, and M. Yanagida Damage and replication checkpoint control in fission yeast is ensured by interactions of Crb2, a protein with BRCT motif, with Cut5 and Chk1 Genes & Dev., December 15, 1997; 11(24): 3387 - 3400. [Abstract] [Full Text] [PDF]

				K. L. Gould and V. Simanis The control of septum formation in fission yeast Genes & Dev., November 15, 1997; 11(22): 2939 - 2951. [Full Text] [PDF]

				H. Mitsuzawa, H. Seino, F. Yamao, and A. Ishihama Two WD Repeat-containing TATA-binding Protein-associated Factors in Fission Yeast That Suppress Defects in the Anaphase-promoting Complex J. Biol. Chem., May 11, 2001; 276(20): 17117 - 17124. [Abstract] [Full Text] [PDF]

				K. M. May, N. Reynolds, C. F. Cullen, M. Yanagida, and H. Ohkura Polo boxes and Cut23 (Apc8) mediate an interaction between polo kinase and the anaphase-promoting complex for fission yeast mitosis J. Cell Biol., January 7, 2002; 156(1): 23 - 28. [Abstract] [Full Text] [PDF]