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A quantitative map of human Condensins provides new insights into mitotic chromosome architecture

View ORCID ProfileNike Walther, View ORCID ProfileM. Julius Hossain, View ORCID ProfileAntonio Z. Politi, Birgit Koch, Moritz Kueblbeck, View ORCID ProfileØyvind Ødegård-Fougner, Marko Lampe, View ORCID ProfileJan Ellenberg
doi: https://doi.org/10.1101/237834
Nike Walther
1Cell Biology & Biophysics Unit, European Molecular Biology Laboratory (EMBL), 69117 Heidelberg, Germany
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M. Julius Hossain
1Cell Biology & Biophysics Unit, European Molecular Biology Laboratory (EMBL), 69117 Heidelberg, Germany
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Antonio Z. Politi
1Cell Biology & Biophysics Unit, European Molecular Biology Laboratory (EMBL), 69117 Heidelberg, Germany
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Birgit Koch
1Cell Biology & Biophysics Unit, European Molecular Biology Laboratory (EMBL), 69117 Heidelberg, Germany
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Moritz Kueblbeck
1Cell Biology & Biophysics Unit, European Molecular Biology Laboratory (EMBL), 69117 Heidelberg, Germany
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Øyvind Ødegård-Fougner
1Cell Biology & Biophysics Unit, European Molecular Biology Laboratory (EMBL), 69117 Heidelberg, Germany
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Marko Lampe
2Advanced Light Microscopy Facility (ALMF), European Molecular Biology Laboratory (EMBL), 69117 Heidelberg, Germany
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Jan Ellenberg
1Cell Biology & Biophysics Unit, European Molecular Biology Laboratory (EMBL), 69117 Heidelberg, Germany
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  • ORCID record for Jan Ellenberg
  • For correspondence: jan.ellenberg@embl.de
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Abstract

The two Condensin complexes in human cells are essential for mitotic chromosome structure. We used homozygous genome editing to fluorescently tag Condensin I and II subunits and mapped their absolute abundance, spacing and dynamic localization during mitosis by fluorescence correlation spectroscopy-calibrated live cell imaging and super-resolution microscopy. While ∼35,000 Condensin II complexes are stably bound to chromosomes throughout mitosis, ∼195,000 Condensin I complexes dynamically bind in two steps, in prometaphase and early anaphase. The two Condensins rarely co-localize at the chromatid axis, where Condensin II is centrally confined but Condensin I reaches ∼50% of the chromatid diameter from its center. Based on our comprehensive quantitative data, we propose a three-step hierarchical loop model of mitotic chromosome compaction: Condensin II initially fixes loops of a maximum size of ∼450 kb at the chromatid axis whose size is then reduced by Condensin I binding to ∼90 kb in prometaphase and ∼70 kb in anaphase, achieving maximum chromosome compaction upon sister chromatid segregation.

  • Abbreviations

    AB
    antibody
    APD
    avalanche photodiode
    CV
    coefficient of variation
    FCS
    fluorescence correlation spectroscopy
    FWHM
    full width at half maximum
    GaAsP
    Gallium arsenide phosphide
    HK
    HeLa Kyoto
    IN
    input
    IP
    immunoprecipitation / eluate
    NEBD
    nuclear envelope breakdown
    Noc
    nocodazole
    ON
    overnight
    POI
    protein of interest
    ROI
    region of interest
    SB
    Southern Blot
    SMC
    structural maintenance of chromosomes
    SN
    supernatant
    STED
    stimulated emission depletion
    WB
    Western Blot
    ZFN
    zinc finger nuclease
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    Posted March 22, 2018.
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    A quantitative map of human Condensins provides new insights into mitotic chromosome architecture
    Nike Walther, M. Julius Hossain, Antonio Z. Politi, Birgit Koch, Moritz Kueblbeck, Øyvind Ødegård-Fougner, Marko Lampe, Jan Ellenberg
    bioRxiv 237834; doi: https://doi.org/10.1101/237834
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    A quantitative map of human Condensins provides new insights into mitotic chromosome architecture
    Nike Walther, M. Julius Hossain, Antonio Z. Politi, Birgit Koch, Moritz Kueblbeck, Øyvind Ødegård-Fougner, Marko Lampe, Jan Ellenberg
    bioRxiv 237834; doi: https://doi.org/10.1101/237834

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