TY - JOUR
T1 - The augmin complex plays a critical role in spindle microtubule generation for mitotic progression and cytokinesis in human cells
AU - Uehara, Ryota
AU - Nozawa, Ryu Suke
AU - Tomioka, Akiko
AU - Petry, Sabine
AU - Vale, Ronald D.
AU - Obuse, Chikashi
AU - Goshima, Gohta
PY - 2009/4/28
Y1 - 2009/4/28
N2 - 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 γ-tubulin ring complex (γ-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 γ-TuRC is likely critical for these functions, because a γ-TuRC mutant that attenuates interaction with augmin does not restore function in vivo. These results demonstrate that MT generation mediated by augmin and γs critical for chromosome segregation and cytokinesis in human cells.
AB - 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 γ-tubulin ring complex (γ-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 γ-TuRC is likely critical for these functions, because a γ-TuRC mutant that attenuates interaction with augmin does not restore function in vivo. These results demonstrate that MT generation mediated by augmin and γs critical for chromosome segregation and cytokinesis in human cells.
KW - Centrosome
KW - Mitosis
KW - RNAi
KW - Spindle checkpoint
UR - http://www.scopus.com/inward/record.url?scp=66349104548&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=66349104548&partnerID=8YFLogxK
U2 - 10.1073/pnas.0901587106
DO - 10.1073/pnas.0901587106
M3 - Article
C2 - 19369198
AN - SCOPUS:66349104548
SN - 0027-8424
VL - 106
SP - 6998
EP - 7003
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
IS - 17
ER -