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Condensin controls mitotic chromosome stiffness and stability without forming a structurally contiguous scaffold

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Abstract

During cell division, chromosomes must be folded into their compact mitotic form to ensure their segregation. This process is thought to be largely controlled by the action of condensin SMC protein complexes on chromatin fibers. However, how condensins organize metaphase chromosomes is not understood. We have combined micromanipulation of single human mitotic chromosomes, sub-nanonewton force measurement, siRNA interference of condensin subunit expression, and fluorescence microscopy, to analyze the role of condensin in large-scale chromosome organization. Condensin depletion leads to a dramatic (~ 10-fold) reduction in chromosome elastic stiffness relative to the native, non-depleted case. We also find that prolonged metaphase stalling of cells leads to overloading of chromosomes with condensin, with abnormally high chromosome stiffness. These results demonstrate that condensin is a main element controlling the stiffness of mitotic chromosomes. Isolated, slightly stretched chromosomes display a discontinuous condensing staining pattern, suggesting that condensins organize mitotic chromosomes by forming isolated compaction centers that do not form a continuous scaffold.

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Abbreviations

(h)CAP:

(human) Chromosome-associated protein

SMC:

Structural maintenance of chromosomes (complex)

siRNA:

Small interfering RNA

WT:

Wild-type (untreated)

NEB:

Nuclear envelope breakdown

CREST:

Calcinosis, Raynaud’s phenomenon, Esophageal dysmotility, Sclerodactyly, and Telangiectasia (syndrome)

CCD:

Charge-coupled device

PBS:

Phosphate-buffered saline

DMEM:

Dulbecco modified eagle medium

FBS:

Fetal bovine serum

Pa:

Pascals (SI unit of pressure)

N:

Newton (SI unit of force)

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Author contribution statement

MS, RB, JH, and JFM conceived and designed the research. MS, RB, and JH conducted experiments. MS, RB, JH, and JFM analyzed data. MS, RB, JH, and JFM wrote, read, and approved the manuscript.

Funding

This work was supported by the NIH through grants R01-GM105847, U54-CA193419 (CR-PS-OC) and a subcontract to grant U54-DK107980, and by the NSF through grants MCB-1022117, DMR-1611076, and DMR-1206868.

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  1. Mingxuan Sun

    Present address: Department of Molecular and Cell Biology, University of California, Berkeley, CA, 94720, USA

Authors and Affiliations

  1. Department of Molecular Biosciences, Northwestern University, Evanston, IL, 60208, USA

    Mingxuan Sun, Ronald Biggs, Jessica Hornick & John F. Marko

  2. Department of Physics and Astronomy, Northwestern University, Evanston, IL, 60208, USA

    John F. Marko

  3. Department of Molecular Biosciences, Northwestern University, 2205 Tech Drive, Evanston, IL, 60208-3500, USA

    John F. Marko

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  1. Mingxuan Sun
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  2. Ronald Biggs
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Correspondence to John F. Marko.

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Responsible Editor: Rachel O’Neill.

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Sun, M., Biggs, R., Hornick, J. et al. Condensin controls mitotic chromosome stiffness and stability without forming a structurally contiguous scaffold. Chromosome Res 26, 277–295 (2018). https://doi.org/10.1007/s10577-018-9584-1

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  • DOI: https://doi.org/10.1007/s10577-018-9584-1

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