Akiyoshi

Bhat

Bishop

Booth

Bouck

Carmena

Coelho

D'Amours

Earnshaw

Fernandes

Gasser

Gassmann

Georgatos

Gerlich

Gibcus

Green

Hagstrom

Hanefeld

Hirano

Hirota

Hochegger

Hori

Houlard

Hudson

Jaqaman

Kakui

Kanemaki

Kruitwagen

Lewis

Llères

Maddox

Mali

Markaki

Martin

Miller

Molnar

Nagasaka

Natsume

Naumova

Nicklas

Nishihashi

Nishimura

Nurse

Ohta

Oliveira

Paulson

Piskadlo

Ribeiro

Saitoh

Saka

Samejima

Samoshkin

Saunders

Smith

Somma

Sonoda

Steffensen

Sullivan

Sutani

Takagi

Vagnarelli

Vassilev

West

Wignall

Wood

Zhiteneva

Edinburgh Research Explorer

Samejima, Kumiko

Booth, Daniel G.

Ogawa, Hiromi

Paulson, James R.

Xie, Linfeng

Watson, Cara A.

Platani, Melpomeni

Kanemaki, Masato T.

Earnshaw, William C.

Repository@Nottingham

Functional analysis after rapid degradation of condensins and 3D-EM reveals chromatin volume is uncoupled from chromosome architecture in mitosis

The requirement for condensin in chromosome formation in somatic cells remains unclear as imperfectly condensed chromosomes do form in conventional condensin-depleted cells. In order to dissect the roles of condensin at different stages of vertebrate mitosis, we have established a versatile cellular system that combines auxin-mediated rapid degradation with chemical genetics to obtain near-synchronous mitotic entry of chicken DT40 cells in the presence and absence of condensin. We analysed the outcome by live and fixed-cell microscopy methods, including serial block face scanning electron microscopy with digital reconstruction. Following rapid depletion of condensin, chromosomal defects were much more obvious compared to those seen after slow depletion of condensin. The total mitotic chromatin volume was similar to that in control cells, but a single mass of mitotic chromosomes was clustered at one side of a bent mitotic spindle. Cultures arrest at prometaphase, eventually exiting mitosis without segregating chromosomes. Experiments titrating the auxin concentration showed that different condensin levels are required for anaphase chromosome segregation and formation of a normal chromosome architecture.</jats:p

Kumiko Samejima

Daniel G. Booth

Hiromi Ogawa

James R. Paulson

Linfeng Xie

Cara A. Watson

Melpomeni Platani

Masato T. Kanemaki

William C. Earnshaw

Crossref

Journal of Cell Science

Rapid degradation of condensins and 3D-EM reveal chromatin volume is uncoupled from chromosome architecture in mitosis

https://www.pure.ed.ac.uk/ws/files/57171704/Functional_analysis_after_rapid_degradation_PUB_OA_CC_BY_jcs210187.full.pdf

Functional analysis after rapid degradation of condensins and 3D-EM reveals chromatin volume is uncoupled from chromosome architecture in mitosis

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Edinburgh Research Explorer

Repository@Nottingham

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