Aurora A contributes to p150(glued) phosphorylation and function during mitosis.
Aurora A is a spindle pole-associated protein kinase required for mitotic spindle assembly and chromosome segregation. In this study, we show that Drosophila melanogaster aurora A phosphorylates the dynactin subunit p150(glued) on sites required for its association with the mitotic spindle. Dynactin strongly accumulates on microtubules during prophase but disappears as soon as the nuclear envelope breaks down, suggesting that its spindle localization is tightly regulated. If aurora A's function is compromised, dynactin and dynein become enriched on mitotic spindle microtubules. Phosphorylation sites are localized within the conserved microtubule-binding domain (MBD) of the p150(glued). Although wild-type p150(glued) binds weakly to spindle microtubules, a variant that can no longer be phosphorylated by aurora A remains associated with spindle microtubules and fails to rescue depletion of endogenous p150(glued). Our results suggest that aurora A kinase participates in vivo to the phosphoregulation of the p150(glued) MBD to limit the microtubule binding of the dynein-dynactin complex and thus regulates spindle assembly.
Romé
Pierre
P
Centre National de la Recherche Scientifique UMR6061, Institut de Génétique et Dévelopement de Rennes, Université de Rennes, 35043 Rennes, France.
Montembault
Emilie
E
Franck
Nathalie
N
Pascal
Aude
A
Glover
David M
DM
Giet
Régis
R
D000818:Animals; D064096:Aurora Kinase A; D064127:Aurora Kinases; D001665:Binding Sites; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000378:metabolism; D000072159:Dynactin Complex; D004398:Dyneins / Q000378:metabolism; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism*; D008870:Microtubules / Q000378:metabolism; D008938:Mitosis; D010766:Phosphorylation; D011494:Protein Kinases / Q000235:genetics / Q000378:metabolism*; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism*; D021122:Protein Subunits / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism; D008941:Spindle Apparatus / Q000378:metabolism
C105393:DCTN1-p150 protein, Drosophila; D029721:Drosophila Proteins; D000072159:Dynactin Complex; D008869:Microtubule-Associated Proteins; D021122:Protein Subunits; D034741:RNA, Small Interfering; D011494:Protein Kinases; D064096:Aurora Kinase A; D064127:Aurora Kinases; D017346:Protein Serine-Threonine Kinases; D004398:Dyneins