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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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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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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DOI: https://doi.org/10.1038/nature07652
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