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


Author(s): Nguyen, Phuong A; Groen, Aaron C; Loose, Martin; Ishihara, Keisuke ; Wühr, Martin ; Field, Christine M; Mitchison, Timothy J
Article Title: Spatial organization of cytokinesis signaling reconstituted in a cell-free system
Affiliation
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.
Keywords: Signal Transduction; Physiology; Xenopus laevis; Genetics; Cytokinesis; chemistry; Cell Membrane; microtubule; metabolism; chromokinesin; DNA binding protein; guanosine triphosphate; kinesin; nuclear protein; RhoA guanine nucleotide binding protein biological model; cell free system; centrosome; lipid bilayer
Journal Title: Science
Volume: 346
Issue 6206
ISSN: 0036-8075
Publisher: American Association for the Advancement of Science  
Date Published: 2014-10-10
Start Page: 244
End Page: 247
Sponsor: This work was supported by NIH grant GM39565 (T.J.M.); MBL fellowships from the Evans Foundation, MBL Associates, and the Colwin Fund (T.J.M. and C.M.F.); HFSP fellowship LT000466/2012-L (M.L.); and NIH grant GM103785 (M.W.).
DOI: 10.1126/science.1256773
Open access: no