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Chromosome Research

, Volume 26, Issue 4, pp 277–295 | Cite as

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

  • Mingxuan Sun
  • Ronald Biggs
  • Jessica Hornick
  • John F. MarkoEmail author
Original Article
  • 453 Downloads

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.

Keywords

metaphase chromosome chromosome compaction condensin colchicine cell division mitosis structural maintenance of chromosome protein chromosome mechanics mechanobiology chromosome stretching non-histone proteins 

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)

Notes

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 information

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.

Supplementary material

10577_2018_9584_MOESM1_ESM.pdf (1.4 mb)
ESM 1 (PDF 1436 kb)

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Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  • Mingxuan Sun
    • 1
    • 2
  • Ronald Biggs
    • 1
  • Jessica Hornick
    • 1
  • John F. Marko
    • 1
    • 3
    • 4
    Email author
  1. 1.Department of Molecular BiosciencesNorthwestern UniversityEvanstonUSA
  2. 2.Department of Molecular and Cell BiologyUniversity of CaliforniaBerkeleyUSA
  3. 3.Department of Physics and AstronomyNorthwestern UniversityEvanstonUSA
  4. 4.Department of Molecular BiosciencesNorthwestern UniversityEvanstonUSA

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