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Spatial organization of cytokinesis signaling reconstituted in a cell-free system

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Science  10 Oct 2014:
Vol. 346, Issue 6206, pp. 244-247
DOI: 10.1126/science.1256773

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Abstract

During animal cell division, the cleavage furrow is positioned by microtubules that signal to the actin cortex at the cell midplane. We developed a cell-free system to recapitulate cytokinesis signaling using cytoplasmic extract from Xenopus eggs. Microtubules grew out as asters from artificial centrosomes and met to organize antiparallel overlap zones. These zones blocked the interpenetration of neighboring asters and recruited cytokinesis midzone proteins, including the chromosomal passenger complex (CPC) and centralspindlin. The CPC was transported to overlap zones, which required two motor proteins, Kif4A and a Kif20A paralog. Using supported lipid bilayers to mimic the plasma membrane, we observed the recruitment of cleavage furrow markers, including an active RhoA reporter, at microtubule overlaps. This system opens further approaches to understanding the biophysics of cytokinesis signaling.

Reconstituting the right stuff for division

Cytokinesis, when two daughter cells are physically separated from one another, is the final stage of cell division. How dividing cells assemble a cleavage furrow ready for cytokinesis has long interested cell biologists. A major stumbling block to probing the underlying mechanisms has been the lack of a cell-free and fully controllable experimental system. Now, Nguyen et al. have reconstituted cytokinesis organization outside living cells, using a system derived from frog eggs. In the cell-free system, the cell cycle state is “frozen,” and the spatial scale is unusually large. The authors examined the biophysics involved in signaling during cytokinesis over many minutes and many micrometers using powerful imaging techniques.

Science, this issue p. 244

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