@article{18dda77f4bd1442bb4586d8784f91f37,
title = "Augmin accumulation on long-lived microtubules drives amplification and kinetochore-directed growth",
abstract = "Dividing cells reorganize their microtubule cytoskeleton into a bipolar spindle, which moves one set of sister chromatids to each nascent daughter cell. Early spindle assembly models postulated that spindle pole-derived microtubules search the cytoplasmic space until they randomly encounter a kinetochore to form a stable attachment. More recent work uncovered several additional, centrosome-independent microtubule generation pathways, but the contributions of each pathway to spindle assembly have remained unclear. Here, we combined live microscopy and mathematical modeling to show that most microtubules nucleate at noncentrosomal regions in dividing human cells. Using a live-cell probe that selectively labels aged microtubule lattices, we demonstrate that the distribution of growing microtubule plus ends can be almost entirely explained by Augmin-dependent amplification of long-lived microtubule lattices. By ultrafast 3D lattice light-sheet microscopy, we observed that this mechanism results in a strong directional bias of microtubule growth toward individual kinetochores. Our systematic quantification of spindle dynamics reveals highly coordinated microtubule growth during kinetochore fiber assembly.",
author = "David, {Ana F.} and Philippe Roudot and Legant, {Wesley R.} and Eric Betzig and Gaudenz Danuser and Gerlich, {Daniel W.}",
note = "Funding Information: Imaging data used in this publication was produced in collaboration with the Advanced Imaging Center, a facility jointly supported by the Gordon and Betty Moore Foundation and Howard Hughes Medical Institute at the Janelia Research Campus. Research in the laboratory of D.W. Gerlich has been supported by the European Community{\textquoteright}s FP7/2007-2013 under grant agreements 241548 (MitoSys) and 258068 (Systems Microscopy), a European Research Council starting grant (281198), the Wiener Wissenschafts-, Forschungs-und Technologiefonds (project no. LS14-009), and the Austrian Science Fund (FWF special research program SFB “Chromosome Dynamics”; project no. SFB F34-06). Research in the laboratory of G. Danuser has been supported by the National Institutes of Health (grant R01 GM067230), and the Human Frontier Science Program (fellowship LT000954/2015 to P. Roudot). The authors declare no competing financial interests. Funding Information: The authors thank Kai Johnsson and Luc Reymond for providing SiR-tubulin and SiR-Hoechst, Grazvydas Lukinavicius and Iva Tolic for comments on the manuscript, IMBA/IMP/GMI Bio-Optics for technical support, and Life Science Editors for editing assistance. Imaging data used in this publication was produced in collaboration with the Advanced Imaging Center, a facility jointly supported by the Gordon and Betty Moore Foundation and Howard Hughes Medical Institute at the Janelia Research Campus. Research in the laboratory of D.W. Gerlich has been supported by the European Community's FP7/2007-2013 under grant agreements 241548 (MitoSys) and 258068 (Systems Microscopy), a European Research Council starting grant (281198), the Wiener Wissenschafts-, Forschungs- und Technologiefonds (project no. LS14-009), and the Austrian Science Fund (FWF special research program SFB ?Chromosome Dynamics?; project no. SFB F34-06). Research in the laboratory of G. Danuser has been supported by the National Institutes of Health (grant R01 GM067230), and the Human Frontier Science Program (fellowship LT000954/2015 to P. Roudot). Publisher Copyright: {\textcopyright} 2019 David et al. This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms/). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 International license, as described at https://creativecommons.org/licenses/by-nc-sa/4.0/).",
year = "2019",
doi = "10.1083/jcb.201805044",
language = "English (US)",
volume = "218",
pages = "2150--2168",
journal = "Journal of Cell Biology",
issn = "0021-9525",
publisher = "Rockefeller University Press",
number = "7",
}