Augmin accumulation on long-lived microtubules drives amplification and kinetochore-directed growth

Ana F. David, Philippe Roudot, Wesley R. Legant, Eric Betzig, Gaudenz Danuser, Daniel W. Gerlich

Research output: Contribution to journalArticle

8 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)2150-2168
Number of pages19
JournalJournal of Cell Biology
Volume218
Issue number7
DOIs
StatePublished - Jan 1 2019

ASJC Scopus subject areas

  • Cell Biology

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