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Mitotic progression, arrest, exit or death is determined by centromere integrity and independent of de novo transcription

View ORCID ProfileMarco Novais-Cruz, Maria Alba Abad, View ORCID ProfileWilfred F.J. van Ijcken, Niels Galjart, View ORCID ProfileA. Arockia Jeyaprakash, View ORCID ProfileHelder Maiato, View ORCID ProfileCristina Ferrás
doi: https://doi.org/10.1101/287151
Marco Novais-Cruz
1Chromosome Instability & Dynamics Laboratory, Instituto de Biologia Molecular e Celular, Universidade do Porto, Rua Alfredo Allen 208, 4200-135 Porto, Portugal
2i3S - Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Rua Alfredo Allen 208, 4200-135 Porto, Portugal
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Maria Alba Abad
3Wellcome Trust Centre for Cell Biology, University of Edinburgh, EH9 3QR, UK
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Wilfred F.J. van Ijcken
4Center for Biomics, Erasmus Medical Center, P.O. Box 2040, 3000 CA Rotterdam, the Netherlands
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Niels Galjart
5Department of Cell Biology, Erasmus Medical Center, P.O. Box 2040, 3000 CA Rotterdam, the Netherlands
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A. Arockia Jeyaprakash
3Wellcome Trust Centre for Cell Biology, University of Edinburgh, EH9 3QR, UK
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Helder Maiato
1Chromosome Instability & Dynamics Laboratory, Instituto de Biologia Molecular e Celular, Universidade do Porto, Rua Alfredo Allen 208, 4200-135 Porto, Portugal
2i3S - Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Rua Alfredo Allen 208, 4200-135 Porto, Portugal
6Cell Division Group, Experimental Biology Unit, Department of Biomedicine, Faculdade de Medicina, Universidade do Porto, Alameda Prof. Hernâni Monteiro, 4200-319 Porto, Portugal
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  • For correspondence: cristina.ferras@ibmc.up.pt maiato@i3s.up.pt
Cristina Ferrás
1Chromosome Instability & Dynamics Laboratory, Instituto de Biologia Molecular e Celular, Universidade do Porto, Rua Alfredo Allen 208, 4200-135 Porto, Portugal
2i3S - Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Rua Alfredo Allen 208, 4200-135 Porto, Portugal
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  • ORCID record for Cristina Ferrás
  • For correspondence: cristina.ferras@ibmc.up.pt maiato@i3s.up.pt
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Abstract

Recent studies have challenged the prevailing dogma that transcription is repressed during mitosis. Transcription was also proposed to sustain the spindle assembly checkpoint (SAC) for several hours in response to unattached kinetochores. Here we used live-cell imaging of human cells in culture, combined with RNA-seq and qPCR, to investigate the requirement for de novo transcription during mitosis. Under conditions of persistently unattached kinetochores, transcription inhibition with actinomycin D, or treatment with other DNA-intercalating drugs, delocalized the chromosomal passenger complex (CPC) protein Aurora B from centromeres, compromising SAC robustness and cell fate. However, we were unable to detect significant changes in transcript levels. Moreover, inhibition of transcription independently of DNA intercalation had no effect on SAC response, mitotic progression, exit or death. Mechanistically, we show that DNA intercalating agents reduce the interaction of the CPC with nucleosomes. Thus, the capacity of human cells to progress, sustain, exit or die in mitosis relies on centromere integrity, rather than de novo transcription.

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Posted March 22, 2018.
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Mitotic progression, arrest, exit or death is determined by centromere integrity and independent of de novo transcription
Marco Novais-Cruz, Maria Alba Abad, Wilfred F.J. van Ijcken, Niels Galjart, A. Arockia Jeyaprakash, Helder Maiato, Cristina Ferrás
bioRxiv 287151; doi: https://doi.org/10.1101/287151
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Mitotic progression, arrest, exit or death is determined by centromere integrity and independent of de novo transcription
Marco Novais-Cruz, Maria Alba Abad, Wilfred F.J. van Ijcken, Niels Galjart, A. Arockia Jeyaprakash, Helder Maiato, Cristina Ferrás
bioRxiv 287151; doi: https://doi.org/10.1101/287151

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