Kinetic framework of spindle assembly checkpoint signaling

Abstract

Author Posting. © The Author(s), 2013. This is the author's version of the work. It is posted here by permission of Nature Publishing Group for personal use, not for redistribution. The definitive version was published in Nature Cell Biology 15 (2013): 1370-1377, doi:10.1038/ncb2842.The mitotic spindle assembly checkpoint (SAC) delays anaphase onset until all chromosomes have attached to both spindle poles1, 2. Here, we investigated SAC signaling kinetics in response to acute detachment of individual chromosomes using laser microsurgery. Most detached chromosomes delayed anaphase until they had realigned to the metaphase plate. A substantial fraction of cells, however, entered anaphase in the presence of unaligned chromosomes. We identify two mechanisms by which cells can bypass the SAC: First, single unattached chromosomes inhibit the anaphase promoting complex/cyclosome (APC/C) less efficiently than a full complement of unattached chromosomes. Second, because of the relatively slow kinetics of reimposing APC/C inhibition during metaphase, cells were unresponsive to chromosome detachment up to several minutes before anaphase onset. Our study defines when cells irreversibly commit to enter anaphase and shows that the SAC signal strength correlates with the number of unattached chromosomes. Detailed knowledge about SAC signaling kinetics is important for understanding the emergence of aneuploidy and the response of cancer cells to chemotherapeutics targeting the mitotic spindle.Research in the Gerlich laboratory has received funding from the European Community’s Seventh Framework Programme FP7/2007-2013 under grant agreements n° 241548 (MitoSys) and n° 258068 (Systems Microscopy), from an ERC Starting Grant (agreement n° 281198), from the EMBO Young Investigator Programme, from the Swiss National Science Foundation, from the Austrian Science Fund (FWF)-funded project “SFB Chromosome Dynamics”, and from a Summer Research Award of the Marine Biology Laboratory Woods Hole (Laura and Arthur Colwin Endowed Summer Research Fellowship Fund). A.E.D. is a fellow of the Zurich Ph.D. Program in Molecular Life Sciences and has received funding from a PhD fellowship by the Boehringer Ingelheim Fonds and from a Peter Müller fellowship.2014-04-0