title: Spatial organization of cytokinesis signaling reconstituted in a cell-free system.
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.
authors.affiliation: Department of Systems Biology, Harvard Medical School, Boston, MA 02115, USA. Marine Biological Laboratory, Woods Hole, MA 02543, USA.
authors.forename: Phuong A
authors.initials: PA
authors.lastname: Nguyen
authors.affiliation: Department of Systems Biology, Harvard Medical School, Boston, MA 02115, USA. Marine Biological Laboratory, Woods Hole, MA 02543, USA.
authors.forename: Aaron C
authors.initials: AC
authors.lastname: Groen
authors.affiliation: Department of Systems Biology, Harvard Medical School, Boston, MA 02115, USA.
authors.forename: Martin
authors.initials: M
authors.lastname: Loose
authors.affiliation: Department of Systems Biology, Harvard Medical School, Boston, MA 02115, USA. Marine Biological Laboratory, Woods Hole, MA 02543, USA.
authors.forename: Keisuke
authors.initials: K
authors.lastname: Ishihara
authors.affiliation: Department of Systems Biology, Harvard Medical School, Boston, MA 02115, USA.
authors.forename: Martin
authors.initials: M
authors.lastname: Wühr
authors.affiliation: Department of Systems Biology, Harvard Medical School, Boston, MA 02115, USA. Marine Biological Laboratory, Woods Hole, MA 02543, USA. Christine_Field@hms.harvard.edu.
authors.forename: Christine M
authors.initials: CM
authors.lastname: Field
authors.affiliation: Department of Systems Biology, Harvard Medical School, Boston, MA 02115, USA. Marine Biological Laboratory, Woods Hole, MA 02543, USA.
authors.forename: Timothy J
authors.initials: TJ
authors.lastname: Mitchison
mesh_terms: D000818:Animals; D002462:Cell Membrane / Q000737:chemistry / Q000502:physiology*; D002474:Cell-Free System; D018385:Centrosome / Q000502:physiology; D048749:Cytokinesis; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D006160:Guanosine Triphosphate / Q000378:metabolism; D016547:Kinesins / Q000235:genetics / Q000378:metabolism; D008051:Lipid Bilayers; D008870:Microtubules / Q000502:physiology; D008954:Models, Biological; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D014982:Xenopus laevis; D020742:rhoA GTP-Binding Protein / Q000378:metabolism
chemical_list: D004268:DNA-Binding Proteins; D008051:Lipid Bilayers; D009687:Nuclear Proteins; C092349:chromokinesin; D006160:Guanosine Triphosphate; D016547:Kinesins; D020742:rhoA GTP-Binding Protein