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Genomic expression patterns in cell separation mutants of Schizosaccharomyces pombe defective in the genes sep10 + and sep15 + coding for the Mediator subunits Med31 and Med8

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Abstract

Cell division is controlled by a complex network involving regulated transcription of genes and postranslational modification of proteins. The aim of this study is to demonstrate that the Mediator complex, a general regulator of transcription, is involved in the regulation of the second phase (cell separation) of cell division of the fission yeast Schizosaccharomyces pombe. In previous studies we have found that the fission yeast cell separation genes sep10 + and sep15 + code for proteins (Med31 and Med8) associated with the Mediator complex. Here, we show by genome-wide gene expression profiling of mutants defective in these genes that both Med8 and Med31 control large, partially overlapping sets of genes scattered over the entire genome and involved in diverse biological functions. Six cell separation genes controlled by the transcription factors Sep1 and Ace2 are among the target genes. Since neither sep1 + nor ace2 + is affected in the mutant cells, we propose that the Med8 and Med31 proteins act as coactivators of the Sep1-Ace2-dependent cell separation genes. The results also indicate that the subunits of Mediator may contribute to the coordination of cellular processes by fine-tuning of the expression of larger sets of genes.

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References

  • Alfa C, Fantes P, Hyams J, McLeod M, Warbrick E (1993) Experiments with fission yeast. A laboratory course manual. Cold Spring Harbor Laboratory, New York

    Google Scholar 

  • Alonzo-Nunez M, An H, Mehta S, Petit C, Sipiczki M, del Rey F, Gould KL, Vazquez de Aldana CR (2005) Ace2 controls the expression of genes required for cell separation in Schizosaccharomyces pombe. Mol Biol Cell 16:2003–2017

    Article  CAS  Google Scholar 

  • Aslett M, Wood V (2006) Gene ontology annotation status of the fission yeast genome: preliminary coverage approaches 100%. Yeast 23:913–919

    Article  PubMed  CAS  Google Scholar 

  • Azad AK, Tani T, Shiki N, Tsuneyoshi S, Urushiyama S, Ohshima Y (1997) Isolation and molecular characterization of mRNA transport mutants in Schizosaccharomyces pombe. Mol Biol Cell 8:825–841

    PubMed  CAS  Google Scholar 

  • Bähler J (2005) A transcriptional pathway for cell separation in fission yeast. Cell Cycle 4:39–41

    PubMed  Google Scholar 

  • Blazek E, Mittler G, Meisterernst M (2005) The mediator of RNA polymerase II. Chromosoma 113:399–408

    Article  PubMed  CAS  Google Scholar 

  • Bourbon H-M et al (2004) A unified nomenclature for protein subunits of Mediator complexes linking transcriptional regulators to RNA polymerase II. Mol Cell 14:553–557

    Article  PubMed  CAS  Google Scholar 

  • Caspari T (1997) Onset of gluconate-H+ symport in Schizosaccharomyces pombe is regulated by the kinases Wis1 and Pka1, and requires the gti1 + gene product. J Cell Sci 110:2599–2608

    PubMed  CAS  Google Scholar 

  • Chaves RS, Herrero P, Moreno F (1999) Med8, a subunit of the mediator CTD complex of RNA polymerase II, directly binds to regulatory elements of SUC2 and HXK2 genes. Biochem Biophys Res Commun 254:345–350

    Article  PubMed  CAS  Google Scholar 

  • Chen D, Toone WM, Mata J, Lyne R, Burns G, Kivinen K, Brazma A, Jones N, Bähler J (2003) Global transcription responses of fission yeast to environmental stress. Mol Biol Cell 14:214–229

    Article  PubMed  CAS  Google Scholar 

  • Cortes JC, Katoh-Fukui R, Moto K, Ribas JC, Ishiguro J (2004) Schizosaccharomyces pombe Pmr1p is essential for cell wall integrity and is required for polarized cell growth and cytokinesis. Eukaryot Cell 3:1124–1135

    Article  PubMed  CAS  Google Scholar 

  • Davis J, Takagi Y, Kornberg R, Asturias F (2002) Structure of the yeast RNA polymerase II holoenzyme. Mediator confirmation and polymerase interaction. Mol Cell 10:409–415

    Article  PubMed  CAS  Google Scholar 

  • Dekker N, Speijer D, Grun CH, van den Berg M, de Haan A, Hochstenbach F (2004) Role of the α-glucanase Agn1p in fission-yeast cell separation. Mol Biol Cell 15:3903–3914

    Article  PubMed  CAS  Google Scholar 

  • Feoktistova A, McCollum D, Ohi R, Gould KL (1999) Identification and characterisation of Schizosaccharomyces pombe asp1 +, a gene that interacts with mutations in the Arp2/3 complex and actin. Genetics 152:895–908

    PubMed  CAS  Google Scholar 

  • Fisher RP (2005) Secrets of a double agent: CDK7 in cell-cycle control and transcription. J Cell Sci 118:5171–5180

    Article  PubMed  CAS  Google Scholar 

  • Forsburg SL (1993) Comparison of Schizosaccharomyces pombe expression systems. Nucleic Acids Res 21:2955–2956

    Article  PubMed  CAS  Google Scholar 

  • Gatti L, Chen D, Beretta GL, Rustici G, Carenini N, Corna E, Colangelo D, Zunino F, Bähler J, Perego P (2004) Global gene expression of fission yeast in response to cisplatin. Cell Mol Life Sci 61:2253–2263

    Article  PubMed  CAS  Google Scholar 

  • Grallert A, Miklos I, Sipiczki M (1997) Division-site selection, cell separation and formation of anucleate minicells in Schizosaccharomyces pombe mutants resistant to cell wall lytic enzymes. Protoplasma 198:218–229

    Article  CAS  Google Scholar 

  • Grallert A, Grallert B, Ribar B, Sipiczki M (1998) Coordination of initiation of nuclear division and initiation of cell division in Schizosaccharomyces pombe: genetic interactions of mutations. J Bacteriol 180:892–900

    PubMed  CAS  Google Scholar 

  • Grallert A, Grallert B, Zilahi E, Szilagyi Z, Sipiczki M (1999) Eleven novel sep genes of Schizosaccharomyces pombe required for efficient cell separation and sexual differentiation. Yeast 15:669–686

    Article  PubMed  CAS  Google Scholar 

  • Guglielmi B, van Berkum N-L, Klapholz B, Bijma T, Boube M, Boschiero C, Bourbon H-M, Holstege FCP, Werner M (2004) A high resolution protein interaction map of the yeast Mediator complex. Nucleic Acids Res 32:5379–5391

    Article  PubMed  CAS  Google Scholar 

  • Hao Z, Furunobu A, Nagata A, Okayama H (1997) A zinc finger protein required for stationary phase viability in fission yeast. J Cell Sci 110:2557–2566

    PubMed  CAS  Google Scholar 

  • Harper JW, Elledge SJ (1998) The role of Cdk7 in CAK function, a retro-retrospective. Genes Dev 12:285–289

    PubMed  CAS  Google Scholar 

  • Heiland S, Radovanovic N, Hofer M, Winderickx J, Lichtenberg H (2000) Multiple hexose transporters of Schizosaccharomyces pombe. J Bacteriol 182:2153–2162

    Article  PubMed  CAS  Google Scholar 

  • Holstege FC, Jennings EG, Wyrick JJ, Lee TI, Hengartner CJ, Green MR, Golub TR, Lander ES, Young RA (1998) Dissecting the regulatory circuitry of a eukaryotic genome. Cell 95:717–728

    Article  PubMed  CAS  Google Scholar 

  • Humbel BM, Konomi M, Takagi T, Kamasawa N, Ishijima SA, Osumi M (2001) In situ localization of β-glucans in the cell wall of Schizosaccharomyces pombe. Yeast 18:433–444

    Article  PubMed  CAS  Google Scholar 

  • Jiang W, Hallberg RL (2001) Correct regulation of the septation initiation network in Schizosaccharomyces pombe requires the activities of par1 and par2. Genetics 158:1413–1429

    PubMed  CAS  Google Scholar 

  • Jiang YW, Stillman DJ (1992) Involvement of the SIN4 global transcriptional regulator in the chromatin structure of Saccharomyces cerevisiae. Mol Cell Biol 12:4503–4514

    PubMed  CAS  Google Scholar 

  • Johnson BF, Yoo BY, Calleja GB (1973) Cell division in yeasts: movement of organelles associated with cell plate growth of Schizosaccharomyces pombe. J Bacteriol 115:358–366

    PubMed  CAS  Google Scholar 

  • Johnson BF, Calleja GB, Zuker M, McDonald TJ (1982) Cell division: key to cellular morphogenesis in the fission yeast Schizosaccharomyces pombe. Int Rev Cytol 75:167–208

    Google Scholar 

  • Kim TW, Kwon Y-J, Kim JM, Song Y-H, Kim SN, Kim Y-J (2004) MED16 and MED23 of Mediator are coactivators of lipopolysaccharide- and heat-shock-induced transcriptional activators. Proc Natl Acad Sci USA 101:12153–12158

    Article  PubMed  CAS  Google Scholar 

  • Lee MK, Miklos I, Du H, Watt S, Szilagyi Z, Saiz JE, Madabhushi R, Penkett CJ, Sipiczki M, Bähler J, Fisher RP (2005) Impairment of the TFIIH-associated CDK-activating kinase selectively affects cell cycle-regulated gene expression in fission yeast. Mol Biol Cell 16:2734–2745

    Article  PubMed  CAS  Google Scholar 

  • Levin HL, Weaver DC, Boeke JD (1990) Two related families of retrotransposons from Schizosaccharomyces pombe. Mol Cell Biol 10:6791–6798

    PubMed  CAS  Google Scholar 

  • Linder T, Gustafsson CM (2004) The Soh1/MED31 protein is an ancient component of Schizosaccharomyces pombe and Saccharomyces cerevisiae Mediator. J Biol Chem 279:49455–49459

    Article  PubMed  CAS  Google Scholar 

  • Lyne R, Burns G, Mata J, Penkett CJ, Rustici G, Chen D, Langford C, Vetrie D, Bähler J (2003) Whole-genome microarrays of fission yeast: characteristics, accuracy, reproducibility, and processing of array data. BMC Genomics 4:27

    Article  PubMed  Google Scholar 

  • Martin-Cuadrado AB, Duenas E, Sipiczki M, Vazquez de Aldana CR, del Rey F (2003) The endo-β-1,3-glucanase Eng1p is required for dissolution of the primary septum during cell separation in Schizosaccharomyces pombe. J Cell Sci 116:1689–1698

    Article  PubMed  CAS  Google Scholar 

  • McCollum D, Balasubramanian MK, Pelcher LE, Hemmingsen SM, Gould KL (1995) Schizosaccharomyces pombe cdc4 + gene encodes a novel EF-hand protein essential for cytokinesis. J Cell Biol 130:651–660

    Article  PubMed  CAS  Google Scholar 

  • Mitchison M (1970) Physiological and cytological methods for Schizosaccharomyces pombe. Methods Cell Physiol 4:131–165

    Google Scholar 

  • Moreno S, Klar A, Nurse P (1991) Molecular genetic analysis of fission yeast Schizosaccharomyces pombe. Methods Enzymol 194:795–823

    Article  PubMed  CAS  Google Scholar 

  • Moreno-Herrero F, Herrero P, Colchero J, Baro AM, Moreno F (1999) Analysis by atomic force microscopy of Med8 binding to cis-acting regulatory elements of the SUC2 and HXK2 genes of Saccharomyces cerevisiae. FEBS Lett 459:427–432

    Article  PubMed  CAS  Google Scholar 

  • Mutoh T, Nakano K, Mabuchi I (2005) Rho1-GEFs Rgf1 and Rgf2 are involved in formation of cell wall and septum, while Rgf3 is involved in cytokinesis in fission yeast. Genes Cells 10:1189–1202

    Article  PubMed  CAS  Google Scholar 

  • Myers LC, Gustafsson CM, Hayashibara KC, Brown PO, Kornberg RD (1999) Mediator protein mutations that selectively abolish activated transcription. Proc Natl Acad Sci USA 96:67–72

    Article  PubMed  CAS  Google Scholar 

  • Nagao K, Adachi Y, Yanagida M (2004) Separase-mediated cleavage of cohesin at interphase is required for DNA repair. Nature 430:1044–1048

    Article  PubMed  CAS  Google Scholar 

  • Palomino A, Herrero P, Moreno F (2005) Rgt1, a glucose sensing transcription factor, is required for transcriptional repression of the HXK2 gene in Saccharomyces cerevisiae. Biochem J 388:697–703

    Article  PubMed  CAS  Google Scholar 

  • Ribar B, Banrevi A, Sipiczki M (1997) sep1 + encodes a transcription-factor homologue of the HNF-3/forkhead DNA-binding-domain family in Schizosaccharomyces pombe. Gene 202:1–5

    Article  PubMed  CAS  Google Scholar 

  • Rustici G, Mata J, Kivinen K, Lio P, Penkett CJ, Burns G, Hayles J, Brazma A, Nurse P, Bähler J (2004) Periodic gene expression program of the fission yeast cell cycle. Nat Genet 36:809–817

    Article  PubMed  CAS  Google Scholar 

  • Saiz JE, Fisher RP (2002) A CDK-activating kinase network is required in cell cycle control and transcription in fission yeast. Curr Biol 12:1100–1105

    Article  PubMed  CAS  Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor

    Google Scholar 

  • Sato S, Suzuki H, Widyastuti U, Hotta Y, Tabata S (1994) Identification and characterization of genes induced during sexual differentiation in Schizosaccharomyces pombe. Curr Genet 26:31–37

    Article  PubMed  CAS  Google Scholar 

  • Seino H, Kishi T, Nishitani H, Yamao F (2003) Two ubiquitin-conjugating enzymes, UbcP1/Ubc4 and UbcP4/Ubc11, have distinct functions for ubiquitination of mitotic cyclin. Mol Cell Biol 23:3497–3505

    Article  PubMed  CAS  Google Scholar 

  • Sharma S, Marguerat S, Mehta S, Watt S, Bähler J (2006) The fission yeast Rpb4 subunit of RNA polymerase II plays a specialized role in cell separation. Mol Genet Genomics 276:545–554

    Article  PubMed  CAS  Google Scholar 

  • Sipiczki M (2007) Splitting of the fission yeast septum. FEMS Yeast Res 7:761–770

    Article  PubMed  CAS  Google Scholar 

  • Sipiczki M, Bozsik A (2000) The use of morphomutants to investigate septum formation and cell separation in Schizosaccharomyces pombe. Arch Microbiol 107:386–392

    Article  Google Scholar 

  • Sipiczki M, Ferenczy L (1977) Protoplast fusion of Schizosaccharomyces pombe auxotrophic mutants of identical mating type. Mol Gen Genet 151:77–81

    Article  PubMed  CAS  Google Scholar 

  • Sipiczki M, Grallert B, Miklos I (1993) Mycelial and syncytial growth in Schizosaccharomyces pombe induced by novel septation mutants. J Cell Sci 104:485–493

    PubMed  Google Scholar 

  • Sipiczki M, Grallert A, Miklos I, Zilahi E, Bozsik A, Szilagyi Z (1999) Genetics, physiology and cytology of yeast-mycelial dimorphism in fission yeasts. Acta Microbiol Immunol Hung 46:297–302

    Article  PubMed  CAS  Google Scholar 

  • Smith DA, Toone WM, Chen D, Bähler J, Jones N, Morgan BA, Quinn J (2002) The Srk1 protein kinase is a target for the Sty1 stress-activated MAPK in fission yeast. J Biol Chem 277:33411–33421

    Article  PubMed  CAS  Google Scholar 

  • Spahr H, Samuelsen CO, Baraznenok V, Ernest I, Huylebroeck D, Remacle J.E, Samuelsson T, Kieselbach T, Holmberg S, Gustafsson CM (2001) Analysis of Schizosaccharomyces pombe mediator reveals a set of essential subunits conserved between yeast and metazoan cells. Proc Natl Acad Sci USA 98:11985–11990

    Article  PubMed  CAS  Google Scholar 

  • Szilagyi Z, Grallert A, Nemeth N, Sipiczki M (2002) The Schizosaccharomyces pombe genes sep10 and sep11 encode putative general transcriptional regulators involved in multiple cellular processes. Mol Genet Genomics 268:553–562

    Article  PubMed  CAS  Google Scholar 

  • Tasto JJ, Morrell JL, Gould KL (2003) An anillin homologue, Mid2p, acts during fission yeast cytokinesis to organize the septin ring and promote cell separation. J Cell Biol 160:1093–1103

    Article  PubMed  CAS  Google Scholar 

  • Thompson JD, Higgins DG, Gibson TJ (1994) CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22:4673–4680

    Article  PubMed  CAS  Google Scholar 

  • Toya M, Iino Y, Yamamoto M (1999) Fission yeast Pob1p, which is homologous to budding yeast Boi proteins and exhibits subcellular localization close to actin patches, is essential for cell elongation and separation. Mol Biol Cell 10:2745–2757

    PubMed  CAS  Google Scholar 

  • Wang L, Griffiths KJ, Zhang YH, Ivey FD, Hoffman CS (2005) Schizosaccharomyces pombe adenylate cyclase suppressor mutations suggest a role for cAMP phosphodiesterase regulation in feedback control of glucose/cAMP signaling. Genetics 171:1523–1533

    Article  PubMed  CAS  Google Scholar 

  • Yamashita YM, Nakaseko Y, Samejima I, Kumada K, Yamada H., Michaelson D, Yanagida M (1996) 20S cyclosome complex formation and proteolytic activity inhibited by the cAMP/PKA pathway. Nature 384:276:279

    Article  PubMed  CAS  Google Scholar 

  • Yang P, Du H, Hoffman CS, Marcus S (2003) The phospholipase B homolog Plb1 is a mediator of osmotic stress response and of nutrient-dependent repression of sexual differentiation in the fission yeast Schizosaccharomyces pombe. Mol Genet Genomics 269:116–125

    PubMed  CAS  Google Scholar 

  • Zilahi E, Miklos I, Sipiczki M (2000) The Schizosaccharomyces pombe sep15 + gene encodes a protein homologues to the Med8 subunit of the Saccharomyces cerevisiae transcriptional mediator complex. Curr Genet 38:227–232

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

We thank Ilona Lakatos for technical assistance. This research was supported by grants from the Hungarian National Fund for Scientific Research (OTKA T042694 and OTKA F48765), by Cancer Research UK [CUK] Grant No. C9546/A6517 (to J.B.), and a Wellcome Trust Short-Term Travel Grant to I.M.

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Authors and Affiliations

  1. Department of Genetics and Applied Microbiology, University of Debrecen, Debrecen, Hungary

    Ida Miklos, Zsolt Szilagyi, Erika Zilahi, Gyula Batta, Zsuzsa Antunovics, Klara Enczi & Matthias Sipiczki

  2. Cancer Research UK Fission Yeast Functional Genomics Group, Wellcome Trust Sanger Institute, Hinxton, Cambridge, CB10 1HH, UK

    Stephen Watt & Jürg Bähler

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  1. Ida Miklos
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  2. Zsolt Szilagyi
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  3. Stephen Watt
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  4. Erika Zilahi
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  5. Gyula Batta
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  7. Klara Enczi
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  8. Jürg Bähler
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  9. Matthias Sipiczki
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Correspondence to Matthias Sipiczki.

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Communicated by S. Hohmann.

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Miklos, I., Szilagyi, Z., Watt, S. et al. Genomic expression patterns in cell separation mutants of Schizosaccharomyces pombe defective in the genes sep10 + and sep15 + coding for the Mediator subunits Med31 and Med8. Mol Genet Genomics 279, 225–238 (2008). https://doi.org/10.1007/s00438-007-0296-z

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