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Cdc14 inhibits transcription by RNA polymerase I during anaphase

Nature volume 458, pages 219–222 (2009)Cite this article

Abstract

Chromosome condensation and the global repression of gene transcription1 are features of mitosis in most eukaryotes. The logic behind this phenomenon is that chromosome condensation prevents the activity of RNA polymerases. In budding yeast, however, transcription was proposed to be continuous during mitosis2. Here we show that Cdc14, a protein phosphatase required for nucleolar segregation3 and mitotic exit4, inhibits transcription of yeast ribosomal genes (rDNA) during anaphase. The phosphatase activity of Cdc14 is required for RNA polymerase I (Pol I) inhibition in vitro and in vivo. Moreover Cdc14-dependent inhibition involves nucleolar exclusion of Pol I subunits. We demonstrate that transcription inhibition is necessary for complete chromosome disjunction, because ribosomal RNA (rRNA) transcripts block condensin binding to rDNA, and show that bypassing the role of Cdc14 in nucleolar segregation requires in vivo degradation of nascent transcripts. Our results show that transcription interferes with chromosome condensation, not the reverse. We conclude that budding yeast, like most eukaryotes, inhibit Pol I transcription before segregation as a prerequisite for chromosome condensation and faithful genome separation.

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Figure 1: Transcription is inhibited in budding yeast during anaphase.
Figure 2: Cdc14 phosphatase inhibits rRNA transcription and prevents binding of Pol I subunits to ribosomal genes.
Figure 3: Cdc14 inhibits RNA Pol I transcription in vitro.
Figure 4: Condensin localization to rDNA and nucleolar segregation requires removal of rRNA transcripts.

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References

  1. Taylor, J. Nucleic acid synthesis in relation to the cell division cycle. Ann. NY Acad. Sci. 90, 409–421 (1960)

    Article  ADS  CAS  Google Scholar 

  2. Elliott, S. G. & McLaughlin, C. S. Regulation of RNA synthesis in yeast. III. Synthesis during the cell cycle. Mol. Gen. Genet. 169, 237–243 (1979)

    Article  CAS  Google Scholar 

  3. Granot, D. & Snyder, M. Segregation of the nucleolus during mitosis in budding and fission yeast. Cell Motil. Cytoskeleton 20, 47–54 (1991)

    Article  CAS  Google Scholar 

  4. Stegmeier, F. & Amon, A. Closing mitosis: the functions of the Cdc14 phosphatase and its regulation. Annu. Rev. Genet. 38, 203–232 (2004)

    Article  CAS  Google Scholar 

  5. Lavoie, B. D., Hogan, E. & Koshland, D. In vivo requirements for rDNA chromosome condensation reveal two cell-cycle-regulated pathways for mitotic chromosome folding. Genes Dev. 18, 76–87 (2004)

    Article  CAS  Google Scholar 

  6. Machin, F., Torres-Rosell, J., Jarmuz, A. & Aragon, L. Spindle-independent condensation-mediated segregation of yeast ribosomal DNA in late anaphase. J. Cell Biol. 168, 209–219 (2005)

    Article  CAS  Google Scholar 

  7. Stegmeier, F., Visintin, R. & Amon, A. Separase, polo kinase, the kinetochore protein Slk19, and Spo12 function in a network that controls Cdc14 localization during early anaphase. Cell 108, 207–220 (2002)

    Article  CAS  Google Scholar 

  8. Machin, F. et al. Transcription of ribosomal genes can cause nondisjunction. J. Cell Biol. 173, 893–903 (2006)

    Article  CAS  Google Scholar 

  9. Tomson, B. N., D’Amours, D., Adamson, B. S., Aragon, L. & Amon, A. Ribosomal DNA transcription-dependent processes interfere with chromosome segregation. Mol. Cell. Biol. 26, 6239–6247 (2006)

    Article  CAS  Google Scholar 

  10. Shou, W. et al. Exit from mitosis is triggered by Tem1-dependent release of the protein phosphatase Cdc14 from nucleolar RENT complex. Cell 97, 233–244 (1999)

    Article  CAS  Google Scholar 

  11. Visintin, R., Hwang, E. S. & Amon, A. Cfi1 prevents premature exit from mitosis by anchoring Cdc14 phosphatase in the nucleolus. Nature 398, 818–823 (1999)

    Article  ADS  CAS  Google Scholar 

  12. Azzam, R. et al. Phosphorylation by cyclin B-Cdk underlies release of mitotic exit activator Cdc14 from the nucleolus. Science 305, 516–519 (2004)

    Article  ADS  CAS  Google Scholar 

  13. Shou, W. et al. Net1 stimulates RNA polymerase I transcription and regulates nucleolar structure independently of controlling mitotic exit. Mol. Cell 8, 45–55 (2001)

    Article  CAS  Google Scholar 

  14. Hendrickson, C., Meyn, M. A., Morabito, L. & Holloway, S. L. The KEN box regulates Clb2 proteolysis in G1 and at the metaphase-to-anaphase transition. Curr. Biol. 11, 1781–1787 (2001)

    Article  CAS  Google Scholar 

  15. Fath, S. et al. Differential roles of phosphorylation in the formation of transcriptional active RNA polymerase I. Proc. Natl Acad. Sci. USA 98, 14334–14339 (2001)

    Article  ADS  CAS  Google Scholar 

  16. Gerber, J. et al. Site specific phosphorylation of yeast RNA polymerase I. Nucleic Acids Res. 36, 793–802 (2008)

    Article  CAS  Google Scholar 

  17. D’Amours, D., Stegmeier, F. & Amon, A. Cdc14 and condensin control the dissolution of cohesin-independent chromosome linkages at repeated DNA. Cell 117, 455–469 (2004)

    Article  Google Scholar 

  18. Sullivan, M., Higuchi, T., Katis, V. L. & Uhlmann, F. Cdc14 phosphatase induces rDNA condensation and resolves cohesin-independent cohesion during budding yeast anaphase. Cell 117, 471–482 (2004)

    Article  CAS  Google Scholar 

  19. Torres-Rosell, J., Machin, F., Jarmuz, A. & Aragon, L. Nucleolar segregation lags behind the rest of the genome and requires Cdc14p activation by the FEAR network. Cell Cycle 3, 496–502 (2004)

    Article  CAS  Google Scholar 

  20. Wang, B. D., Yong-Gonzalez, V. & Strunnikov, A. V. Cdc14p/FEAR pathway controls segregation of nucleolus in S. cerevisiae by facilitating condensin targeting to rDNA chromatin in anaphase. Cell Cycle 3, 960–967 (2004)

    CAS  PubMed  PubMed Central  Google Scholar 

  21. Wang, B. D., Butylin, P. & Strunnikov, A. Condensin function in mitotic nucleolar segregation is regulated by rDNA transcription. Cell Cycle 5, 2260–2267 (2006)

    Article  CAS  Google Scholar 

  22. Fujii, T., Yamaoka, H., Gomi, K., Kitamoto, K. & Kumagai, C. Cloning and nucleotide sequence of the ribonuclease T1 gene (rntA) from Aspergillus oryzae and its expression in Saccharomyces cerevisiae and Aspergillus oryzae. Biosci. Biotechnol. Biochem. 59, 1869–1874 (1995)

    Article  CAS  Google Scholar 

  23. D’Ambrosio, C., Kelly, G., Shirahige, K. & Uhlmann, F. Condensin-dependent rDNA decatenation introduces a temporal pattern to chromosome segregation. Curr. Biol. 18, 1084–1089 (2008)

    Article  Google Scholar 

  24. Freeman, L., Aragon-Alcaide, L. & Strunnikov, A. The condensin complex governs chromosome condensation and mitotic transmission of rDNA. J. Cell Biol. 149, 811–824 (2000)

    Article  CAS  Google Scholar 

  25. Baxter, J. & Diffley, J. F. Topoisomerase II inactivation prevents the completion of DNA replication in budding yeast. Mol. Cell 30, 790–802 (2008)

    Article  CAS  Google Scholar 

  26. Bhalla, N., Biggins, S. & Murray, A. W. Mutation of YCS4, a budding yeast condensin subunit, affects mitotic and nonmitotic chromosome behavior. Mol. Biol. Cell 13, 632–645 (2002)

    Article  CAS  Google Scholar 

  27. Nelson, J. D., Denisenko, O. & Bomsztyk, K. Protocol for the fast chromatin immunoprecipitation (ChIP) method. Nature Protocols 1, 179–185 (2006)

    Article  CAS  Google Scholar 

  28. Keener, J., Josaitis, C. A., Dodd, J. A. & Nomura, M. Reconstitution of yeast RNA polymerase I transcription in vitro from purified components. TATA-binding protein is not required for basal transcription. J. Biol. Chem. 273, 33795–33802 (1998)

    Article  CAS  Google Scholar 

  29. Janke, C. et al. A versatile toolbox for PCR-based tagging of yeast genes: new fluorescent proteins, more markers and promoter substitution cassettes. Yeast 21, 947–962 (2004)

    Article  CAS  Google Scholar 

  30. Powers, T. & Walter, P. Regulation of ribosome biogenesis by the rapamycin-sensitive TOR-signaling pathway in Saccharomyces cerevisiae. Mol. Biol. Cell 10, 987–1000 (1999)

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We are grateful to A. Amon, D. Morgan, K. Kitamoto and J. Van Etten for providing reagents, and M. Nomura for advice. We thank members of the Aragon laboratory for discussions. We acknowledge grant support from UAB HSF-GEF to D.A.S. and from ‘La Junta de Extremadura’ to A.C.-B. The Aragon laboratory is supported by the Medical Research Council of the UK.

Author contributions F.M. made the initial observation that transcription impairs rDNA segregation in the absence of Cdc14 function. All experiments shown were performed by A.C.-B., except ChIPs which were performed by M.M.-S. and in vitro phosphatase assays which were performed by D.A.S. A.J. cloned the RNAse constructs. H.T. provided a battery of Rpa43 mutants. All experiments shown were conceived by L.A. and the person performing the specific experiment. L.A. wrote the paper.

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  1. Félix Machín

    Present address: Present address: Unidad de Investigación, Hospital Universitario Candelaria, Carretera del Rosario, 38010 Tenerife, Spain.,

Authors and Affiliations

  1. Cell Cycle Group, MRC Clinical Sciences Centre, Imperial College, Du Cane Road, London W12 0NN, UK,

    Andrés Clemente-Blanco, María Mayán-Santos, Félix Machín, Adam Jarmuz & Luis Aragón

  2. Department of Biochemistry and Molecular Genetics, University of Alabama at Birmingham, 442 Kaul Human Genetics Building, 720 20th Street South, Birmingham, Alabama 35294, USA,

    David A. Schneider

  3. Institut für Biochemie, Mikrobiologie und Genetik, Universität Regensburg, Munich, Germany

    Herbert Tschochner

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Correspondence to Luis Aragón.

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Clemente-Blanco, A., Mayán-Santos, M., Schneider, D. et al. Cdc14 inhibits transcription by RNA polymerase I during anaphase. Nature 458, 219–222 (2009). https://doi.org/10.1038/nature07652

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