Accessible Requires Authentication Published by De Gruyter August 8, 2005

Psc3 cohesin of Schizosaccharomyces pombe: cell cycle analysis and identification of three distinct isoforms

Eduard Ilyushik, David W. Pryce, Dawid Walerych, Tracy Riddell, Jane A. Wakeman, Christopher J. McInerny and Ramsay J. McFarlane
From the journal

Abstract

Cohesins are a group of proteins that function to mediate correct chromosome segregation, DNA repair and meiotic recombination. This report presents the amino acid sequence for the Schizosaccharomyces pombe cohesin Psc3 based on the translation of the cDNA sequence, showing that the protein is smaller than previously predicted. Interestingly, comparison of the amino acid and DNA coding sequences of Psc3 with fission yeast Rec11 meiotic region-specific recombination activator shows that both intron positioning within the genes and the amino-terminal half of the two proteins are highly conserved. We demonstrate that although the intergenic region upstream of the psc3+ start codon contains a consensus sequence for the cell-cycle regulatory MluI cell-cycle box, psc3+ transcription is not differentially regulated during the mitotic cell cycle. Finally, we demonstrate that an epitope-tagged version of Psc3 undergoes no major changes during the mitotic cell cycle. However, instead we identify at least three distinct isoforms of Psc3, suggesting that post-translational modification of Psc3 contributes to the regulation of cohesion function.

:

Corresponding author

References

Amon, A. (2001). Together until separin do us part. Nat. Cell Biol.3, E12–E14. Search in Google Scholar

Bähler, J., Wu, J.-Q., Longtine, M.S., Shah, N.G., McKenzie, A., Steever, A.B., Wach, A., Philippsen, P., and Pringle, J.R. (1998). Heterologous modules for efficient and versatile PCR-based gene targeting in Schizosaccharomyces pombe. Yeast14, 943–951. Search in Google Scholar

Baum, B., Wuarin, J., and Nurse, P. (1997). Control of S-phase transcription in the fission yeast mitotic cycle. EMBO J.16, 4676–4688. Search in Google Scholar

Birkenbihl, R.P. and Subramani, S. (1992). Cloning and characterization of rad21 an essential gene of Schizosaccharomyces pombe involved in DNA double-strand-break repair. Nucleic Acids Res.20, 6605–6611. Search in Google Scholar

Birkenbihl, R.P. and Subramani, S. (1995). The rad21 gene product of Schizosaccharomyces pombe is a nuclear, cell cycle regulated phosphoprotein. J. Biol. Chem.270, 7703–7711. Search in Google Scholar

Davis, L. and Smith, G.R. (2001). Meiotic recombination and chromosome segregation in Schizosaccharomyces pombe.Proc. Natl. Acad. Sci. USA98, 8395–8402. Search in Google Scholar

DeVeaux, L.C. and Smith, G.R. (1994). Region-specific activators of meiotic recombination in Schizosaccharomyces pombe.Genes Dev.8, 203–210. Search in Google Scholar

Feinberg, A.P. and Volgelstein, B. (1983). A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal. Biochem.1, 6–13. Search in Google Scholar

Hagstrom, K.A. and Meyer, B.J. (2003). Condensin and cohesion: more than chromosome compactor and glue. Nat. Rev. Genet.4, 520–534. Search in Google Scholar

Hauf, S., Waizenegger, I.C., and Peters, J.-M. (2001). Cohesin cleavage by separase required for anaphase and cytokinesis in human cells. Science293, 1320–1323. Search in Google Scholar

Hauf, S., Roitinger, E., Dittrich, C.M., Mechtler, K., and Peters, J.-M. (2005). Dissociation of cohesin from chromosome arms and loss of arm cohesion during early mitosis depends on phosphorylation. PLoS Biol.3, 419–432. Search in Google Scholar

Hiraoka, Y., Toda, T., and Yanagida, M. (1984). The NDA3 gene of fission yeast encodes beta-tubulin: a cold-sensitive nda3 mutation reversibly blocks spindle formation and chromosome movement in mitosis. Cell39, 349–358. Search in Google Scholar

Hofmann, J.F.X. and Beach D. (1994). cdt1 is an essential target of the Cdc10/Sct1 transcription factor: requirement for DNA replication and inhibition of mitosis. EMBO J.13, 425–434. Search in Google Scholar

Hoving, S., Voshol, H., and Oostrum, J. (2000). Towards high performance two-dimensional gel electrophoresis using ultrazoom gels. Electrophoresis21, 2617–2621. Search in Google Scholar

Ivanov, D., Schleiffer, A., Eisenhaber, F., Mechtler, K., Haering, C.H., and Nasmyth, K. (2002). Eco1 is a novel acetyltransferase that can acetylate proteins involved in cohesion. Curr. Biol.12, 323–328. Search in Google Scholar

Jones, S. and Sgouros, J. (2001). The cohesin complex: sequence homologies, interaction networks and shared motifs. Genome Biol.2, 1–12. Search in Google Scholar

Krawchuk, M.D., DeVeaux, L.C., and Wahls, W.P. (1999). Meiotic chromosome dynamics dependent upon the rec8+, rec10+ and rec11+ genes of the fission yeast Schizosaccharomyces pombe. Genetics153, 57–68. Search in Google Scholar

Lara-Pezzi, E., Pezzi, N., Prieto, I., Barthelemy, I., Carreiro, C., Martínez, A., Maldonado-Radríguez, A., López-Cabrera, M., and Barbero, J.L. (2004). Evidence of a transcriptional co-activator function of cohesin STAG/SA/Scc3. J. Biol. Chem.279, 6553–6559. Search in Google Scholar

Li, Y.F., Numata, M., Wahls, W.P., and Smith, G.R. (1997). Region-specific meiotic recombination in Schizosaccharomyces pombe: the rec11 gene. Mol. Microbiol.23, 869–878. Search in Google Scholar

Lowndes, N.F., McInerny, C.J., Johnson, A.L., Fantes, P.A., and Johnston, L.H. (1992). Control of DNA synthesis genes in fission yeast by the cell cycle gene cdc10+. Nature355, 449–455. Search in Google Scholar

McInerny, C.J., Kersey, P.J., Creanor, J., and Fantes, P.A. (1995). Positive and negative roles for cdc10 in cell cycle gene expression. Nucleic Acids Res.23, 4761–4768. Search in Google Scholar

Michaelis, C., Ciosk, R., and Nasmyth, K. (1997). Cohesins: chromosomal proteins that prevent premature separation of sister chromatids. Cell91, 35–45. Search in Google Scholar

Moreno, S., Klar, A., and Nurse, P. (1991). Molecular genetic analysis of fission yeast Schizosaccharomyces pombe. Methods Enzymol.194, 795–823. Search in Google Scholar

Nagao, K., Adachi, Y., and Yanagida, M. (2004). Separase mediated cleavage of cohesin at interphase is required for DNA repair. Nature430, 1044–1048. Search in Google Scholar

Nasmyth, K. (2002). Segregating sister genomes: the molecular biology of chromosome separation. Science297, 559–565. Search in Google Scholar

Nonaka, N., Kitajima, T., Yokobayashi, S., Xiao, G., Yamamoto, M., Grewal, S.I., and Watanabe, Y. (2002). Recruitment of cohesion to heterochromatic regions by Swi6/HP1 in fission yeast. Nat. Cell Biol.4, 89–93. Search in Google Scholar

Rechsteiner, M. and Rogers, S.W. (1996). PEST sequences and regulation by proteolysis. Trends Biochem. Sci.21, 267–271. Search in Google Scholar

Rogers, S., Wells, R., and Rechsteiner, M. (1986). Amino acid sequences common to rapidly degraded proteins: the PEST hypothesis. Science234, 364–368. Search in Google Scholar

Sambrook, J., Fritsch, E.F., and Maniatis, T. (1989). Molecular Cloning: A Laboratory Manual, 2nd Edition (Cold Spring Harbor, USA: Cold Spring Harbor Laboratory Press). Search in Google Scholar

Sjogren, C. and Nasmyth, K. (2001). Sister chromatid cohesion is required for postreplicative double-strand break repair in Saccharomyces cerevisiae. Curr. Biol.11, 991–995. Search in Google Scholar

Tomonaga, T., Nagao, K., Kawaasaki, Y., Furuya, K., Murakami, A., Morishita, J., Yuasa, T., Sutani, T., Kearsey, S.E., Uhlmann, F., Nasmyth, K., and Yanagida, M. (2000). Characterization of fission yeast cohesin: essential anaphase proteolysis of Rad21 phosphorylated in the S phase. Genes Dev.14, 2757–2770. Search in Google Scholar

Toth, A., Ciosk, R., Uhlmann, F., Galova, M., Schleiffer, A., and Nasmyth, K. (1999). Yeast cohesin complex requires a conserved protein, Eco1p (Ctf7), to establish cohesion between sister chromatids during DNA replication. Genes Dev.13, 320–333. Search in Google Scholar

Ulmann, F. (2004). The mechanism of sister chromatid cohesion. Exp. Cell Res.296, 80–85. Search in Google Scholar

Whitehall, S., Stacey, P., Dawson, K., and Jones, N. (1999). Cell cycle-regulated transcription in fission yeast: Cdc10-Res protein interactions during the cell cycle and domains required for regulated transcription. Mol. Biol. Cell10, 3705–3715. Search in Google Scholar

Wood, V., Gwilliam, R., Rajandream, M.A., Lyne, M., Lyne, R., Stewart, A., Sgouros, J., Peat, N., Hayles, J., Baker, S., et al. (2002). The genome sequence of Schizosaccharomyces pombe. Nature415, 871–880. Search in Google Scholar

Zhang, M.Q. and Marr, T.G. (1994). Fission yeast gene structure and recognition. Nucleic Acids Res.22, 1750–1759. Search in Google Scholar

Published Online: 2005-08-08
Published in Print: 2005-07-01

©2005 by Walter de Gruyter Berlin New York