期刊
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
卷 106, 期 17, 页码 6998-7003出版社
NATL ACAD SCIENCES
DOI: 10.1073/pnas.0901587106
关键词
centrosome; mitosis; RNAi; spindle checkpoint
资金
- Ministry of Education, Culture, Sports, Science and Technology (Japan)
- Nakajima Foundation
- Uehara Foundation
- Cell Science Foundation
- Human Frontier Science Program
- Global Centers of Excellence program
- European Molecular Biology Organization postdoctoral fellowship
- National Institutes of Health [38499]
- Grants-in-Aid for Scientific Research [20114006] Funding Source: KAKEN
The mitotic spindle is constructed from microtubules (MTs) nucleated from centrosomes, chromosome proximal regions, and preexisting spindle MTs. Augmin, a recently identified protein complex, is a critical factor in spindle MT-based MT generation in Drosophila S2 cells. Previously, we identified one subunit of human augmin. Here, by using mass spectrometry, we identified the full human augmin complex of 8 subunits and show that it interacts with the gamma-tubulin ring complex (gamma-TuRC). Unlike augmin-depleted S2 cells, in which the defect in spindle-mediated MT generation is mostly compensated by centrosomal MTs, augmin knockdown alone in HeLa cells triggers the spindle checkpoint, reduces tension on sister kinetochores, and severely impairs metaphase progression. Human augmin knockdown also reduces the number of central spindle MTs during anaphase and causes late-stage cytokinesis failure. A link between augmin and gamma-TuRC is likely critical for these functions, because a gamma-TuRC mutant that attenuates interaction with augmin does not restore function in vivo. These results demonstrate that MT generation mediated by augmin and gamma-TuRC is critical for chromosome segregation and cytokinesis in human cells.
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