Isolation of a Schizosaccharomyces pombe rad21^ts Mutant That Is Aberrant in Chromosome Segregation, Microtubule Function, DNA Repair and Sensitive to Hydroxyurea: Possible Involvement of Rad21 in Ubiquitin-Mediated Proteolysis

Corresponding author: Hideo Ikeda, Department of Molecular Biology, Institute of Medical Science, University of Tokyo, 4-6-1, Shirokanedai Minato-ku, P.O. Takanawa, Tokyo 108, Japan, ike{at}hgc.ims.u-tokyo.ac.jp (E-mail).

Communicating editor: F. WINSTON

The fission yeast DNA repair gene rad21⁺ is essential for cell growth. To investigate the function essential for cell proliferation, we have isolated a temperature-sensitive mutant of the rad21⁺ gene. The mutant, rad21-K1, showed abnormal mitosis at the nonpermissive temperature. Some cells contained abnormal nuclear structures, such as condensed chromosomes with short spindles, or chromosomes stretched or unequally separated by elongating spindles. Other cells exhibited the displaced nucleus or a cut-like phenotype. Similar abnormalities were observed when the Rad21 protein was depleted from cells. We therefore concluded that Rad21 is essential for proper segregation of chromosomes. Moreover, the rad21-K1 mutant is sensitive not only to UV and -ray irradiation but to thiabendazole and hydroxyurea, indicating that Rad21 plays important roles in microtubule function, DNA repair, and S phase function. The relation to the microtubule function was further confirmed by the fact that rad21⁺ genetically interacts with tubulin genes, nda2⁺ and nda3 ⁺. Finally, the growth of the rad21-K1 mutant was inhibited at the permissive temperature by introduction of another mutation in the cut9 ⁺ gene, coding for a component of the 20S cyclosome/anaphase promoting complex, which is involved in ubiquitin-mediated proteolysis. The results suggest that these diverse functions of Rad21 may be facilitated through ubiquitin-mediated proteolysis.

This article has been cited by other articles:

				Y. Tange and O. Niwa Schizosaccharomyces pombe Bub3 Is Dispensable for Mitotic Arrest Following Perturbed Spindle Formation Genetics, June 1, 2008; 179(2): 785 - 792. [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]

				A. B. Ansbach, C. Noguchi, I. W. Klansek, M. Heidlebaugh, T. M. Nakamura, and E. Noguchi RFCCtf18 and the Swi1-Swi3 Complex Function in Separate and Redundant Pathways Required for the Stabilization of Replication Forks to Facilitate Sister Chromatid Cohesion in Schizosaccharomyces pombe Mol. Biol. Cell, February 1, 2008; 19(2): 595 - 607. [Abstract] [Full Text] [PDF]

				K. M. Lee, S. Nizza, T. Hayes, K. L. Bass, A. Irmisch, J. M. Murray, and M. J. O'Connell Brc1-Mediated Rescue of Smc5/6 Deficiency: Requirement for Multiple Nucleases and a Novel Rad18 Function Genetics, April 1, 2007; 175(4): 1585 - 1595. [Abstract] [Full Text] [PDF]

				K. Asakawa, K. Kume, M. Kanai, T. Goshima, K. Miyahara, S. Dhut, W. W. Tee, D. Hirata, and T. Toda The V260I Mutation in Fission Yeast {alpha}-Tubulin Atb2 Affects Microtubule Dynamics and EB1-Mal3 Localization and Activates the Bub1 Branch of the Spindle Checkpoint Mol. Biol. Cell, March 1, 2006; 17(3): 1421 - 1435. [Abstract] [Full Text] [PDF]

				S. W. Bai, J. Rouquette, M. Umeda, W. Faigle, D. Loew, S. Sazer, and V. Doye The Fission Yeast Nup107-120 Complex Functionally Interacts with the Small GTPase Ran/Spi1 and Is Required for mRNA Export, Nuclear Pore Distribution, and Proper Cell Division Mol. Cell. Biol., July 15, 2004; 24(14): 6379 - 6392. [Abstract] [Full Text] [PDF]

				S. H. Harvey, D. M. Sheedy, A. R. Cuddihy, and M. J. O'Connell Coordination of DNA Damage Responses via the Smc5/Smc6 Complex Mol. Cell. Biol., January 15, 2004; 24(2): 662 - 674. [Abstract] [Full Text] [PDF]

				S. Yokobayashi, M. Yamamoto, and Y. Watanabe Cohesins Determine the Attachment Manner of Kinetochores to Spindle Microtubules at Meiosis I in Fission Yeast Mol. Cell. Biol., June 1, 2003; 23(11): 3965 - 3973. [Abstract] [Full Text] [PDF]

				Y. Mito, A. Sugimoto, and M. Yamamoto Distinct Developmental Function of Two Caenorhabditis elegans Homologs of the Cohesin Subunit Scc1/Rad21 Mol. Biol. Cell, June 1, 2003; 14(6): 2399 - 2409. [Abstract] [Full Text] [PDF]

				Md. T. Hoque and F. Ishikawa Cohesin Defects Lead to Premature Sister Chromatid Separation, Kinetochore Dysfunction, and Spindle-assembly Checkpoint Activation J. Biol. Chem., October 25, 2002; 277(44): 42306 - 42314. [Abstract] [Full Text] [PDF]

				D. R. Williams and J. R. McIntosh mcl1+, the Schizosaccharomyces pombe Homologue of CTF4, Is Important for Chromosome Replication, Cohesion, and Segregation Eukaryot. Cell, October 1, 2002; 1(5): 758 - 773. [Abstract] [Full Text] [PDF]

				T. Morishita, Y. Tsutsui, H. Iwasaki, and H. Shinagawa The Schizosaccharomyces pombe rad60 Gene Is Essential for Repairing Double-Strand DNA Breaks Spontaneously Occurring during Replication and Induced by DNA-Damaging Agents Mol. Cell. Biol., May 15, 2002; 22(10): 3537 - 3548. [Abstract] [Full Text] [PDF]

				F. Chen, M. Kamradt, M. Mulcahy, Y. Byun, H. Xu, M. J. McKay, and V. L. Cryns Caspase Proteolysis of the Cohesin Component RAD21 Promotes Apoptosis J. Biol. Chem., May 3, 2002; 277(19): 16775 - 16781. [Abstract] [Full Text] [PDF]

				T. Takeda, K. Ogino, K. Tatebayashi, H. Ikeda, K.-i. Arai, and H. Masai Regulation of Initiation of S Phase, Replication Checkpoint Signaling, and Maintenance of Mitotic Chromosome Structures during S Phase by Hsk1 Kinase in the Fission Yeast Mol. Biol. Cell, May 1, 2001; 12(5): 1257 - 1274. [Abstract] [Full Text]

				K. Tatebayashi, T. Tani, and H. Ikeda Fission Yeast Mog1p Homologue, Which Interacts With the Small GTPase Ran, Is Required for Mitosis-to-Interphase Transition and poly(A)+ RNA Metabolism Genetics, April 1, 2001; 157(4): 1513 - 1522. [Abstract] [Full Text]

				T. Shobuike, K. Tatebayashi, T. Tani, S. Sugano, and H. Ikeda The dhp1+ gene, encoding a putative nuclear 5'{->}3' exoribonuclease, is required for proper chromosome segregation in fission yeast Nucleic Acids Res., March 15, 2001; 29(6): 1326 - 1333. [Abstract] [Full Text] [PDF]

				D. J. F. Griffiths, V. F. Liu, P. Nurse, and T. S.-F. Wang Role of Fission Yeast Primase Catalytic Subunit in the Replication Checkpoint Mol. Biol. Cell, January 1, 2001; 12(1): 115 - 128. [Abstract] [Full Text]

				S. Tan and T. S.-F. Wang Analysis of Fission Yeast Primase Defines the Checkpoint Responses to Aberrant S Phase Initiation Mol. Cell. Biol., November 1, 2000; 20(21): 7853 - 7866. [Abstract] [Full Text]

				H. A. Snaith, G. W. Brown, and S. L. Forsburg Schizosaccharomyces pombe Hsk1p Is a Potential Cds1p Target Required for Genome Integrity Mol. Cell. Biol., November 1, 2000; 20(21): 7922 - 7932. [Abstract] [Full Text]

				T. Tomonaga, K. Nagao, Y. Kawasaki, K. Furuya, A. Murakami, J. Morishita, T. Yuasa, T. Sutani, S. E. Kearsey, F. Uhlmann, et al. Characterization of fission yeast cohesin: essential anaphase proteolysis of Rad21 phosphorylated in the S phase Genes & Dev., November 1, 2000; 14(21): 2757 - 2770. [Abstract] [Full Text]

				K. Takahashi, E. S. Chen, and M. Yanagida Requirement of Mis6 Centromere Connector for Localizing a CENP-A-Like Protein in Fission Yeast Science, June 23, 2000; 288(5474): 2215 - 2219. [Abstract] [Full Text]

				K. Tanaka, T. Yonekawa, Y. Kawasaki, M. Kai, K. Furuya, M. Iwasaki, H. Murakami, M. Yanagida, and H. Okayama Fission Yeast Eso1p Is Required for Establishing Sister Chromatid Cohesion during S Phase Mol. Cell. Biol., May 15, 2000; 20(10): 3459 - 3469. [Abstract] [Full Text]

				M. D. Krawchuk, L. C. DeVeaux, and W. P. Wahls Meiotic Chromosome Dynamics Dependent Upon the rec8+, rec10+ and rec11+ Genes of the Fission Yeast Schizosaccharomyces pombe Genetics, September 1, 1999; 153(1): 57 - 68. [Abstract] [Full Text]

				H. M. Verkade, S. J. Bugg, H. D. Lindsay, A. M. Carr, and M. J. O'Connell Rad18 Is Required for DNA Repair and Checkpoint Responses in Fission Yeast Mol. Biol. Cell, September 1, 1999; 10(9): 2905 - 2918. [Abstract] [Full Text]

				T. Hirano SMC-mediated chromosome mechanics: a conserved scheme from bacteria to vertebrates? Genes & Dev., January 1, 1999; 13(1): 11 - 19. [Full Text]