Abstract
The dynactin complex cooperates with the dynein complex in various systems for mitotic completion. Here we analysed the mitotic phenotype of Drosophila S2 cells following the knockdown of the dynactin subunit p150(Glued). We found that p150(Glued)-depleted cells were delayed in metaphase and that the centrosomes were poorly connected to mitotic spindle poles. In addition, anaphase occurred with asynchronous chromosome segregation. Although cyclin B was degraded in these anaphase cells, Aurora B, MEI-S322 and BubR1 were not released from the non-segregating chromosomes. We also found that the density and organisation of the central spindle were compromised, with Aurora B and polo kinases absent from the diminished number of microtubules. Pavarotti-KLP, a component of the centralspindlin complex required for the formation of stable microtubule bundles, was not immediately targeted to the plus ends of the microtubules following anaphase onset as happened in controls. Instead, it accumulated transiently at the cell cortex during early anaphase and its targeting to the central spindle was delayed. These data suggest that the dynactin complex contributes to cytokinesis by promoting stable targeting of the centralspindlin complex to microtubule plus ends at anaphase onset. The contribution of the dynein-dynactin complex to synchronous chromosome segregation and cytokinesis is discussed.
Highlights
Dynein is a microtubule motor that moves toward the minus end of microtubules as part of a large macromolecular complex (Collins and Vallee, 1989; Vallee et al, 1989)
We consistently found that cyclin B was degraded in Glued-depleted cells with elongated spindles and aberrantly positioned chromosomes (Fig. 4A, right panels), indicating that these cells had progressed into anaphase
Contribution of dynein-dynactin complex (DDC) to synchronous chromosome segregation Here we show that in Drosophila cultured cells, knockdown of p150Glued leads to a metaphase arrest and after the eventual transition into anaphase, disrupts chromosome segregation and central spindle formation
Summary
Dynein is a microtubule motor that moves toward the minus end of microtubules as part of a large macromolecular complex (Collins and Vallee, 1989; Vallee et al, 1989). Cytoplasmic dynein, which is present at the cell cortex, pulls on centrosomeconnected microtubules and is responsible for the positioning of the centrosomes in the centre of the cell (Koonce et al, 1999; Vallee and Stehman, 2005). It has been suggested that one major function of dynactin is to target dynein where it is required in the cell (Vallee et al, 1995)
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