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Vol. 16, Issue 7, 3176-3186, July 2005

This Article Full Text Full Text (PDF) Supplemental Material A correction has been published All Versions of this Article: E04-12-1110v1 E04-12-1110v2 16/7/3176    most recent Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Morales-Mulia, S. Articles by Scholey, J. M. Search for Related Content PubMed PubMed Citation Articles by Morales-Mulia, S. Articles by Scholey, J. M.

Spindle Pole Organization in Drosophila S2 Cells by Dynein, Abnormal Spindle Protein (Asp), and KLP10A

Section of Molecular and Cellular Biology, Center for Genetics and Development, University of California–Davis, Davis, CA 95616

Submitted December 23, 2004; Accepted April 27, 2005
Monitoring Editor: Yixian Zheng

Dynein is a critical mitotic motor whose inhibition causes defects in spindle pole organization and separation, chromosome congression or segregation, and anaphase spindle elongation, but results differ in different systems. We evaluated the functions of the dynein–dynactin complex by using RNA interference (RNAi)-mediated depletion of distinct subunits in Drosophila S2 cells. We observed a striking detachment of centrosomes from spindles, an increase in spindle length, and a loss of spindle pole focus. RNAi depletion of Ncd, another minus-end motor, produced disorganized spindles consisting of multiple disconnected mini-spindles, a different phenotype consistent with distinct pathways of spindle pole organization. Two candidate dynein-dependent spindle pole organizers also were investigated. RNAi depletion of the abnormal spindle protein, Asp, which localizes to focused poles of control spindles, produced a severe loss of spindle pole focus, whereas depletion of the pole-associated microtubule depolymerase KLP10A increased spindle microtubule density. Depletion of either protein produced long spindles. After RNAi depletion of dynein–dynactin, we observed subtle but significant mislocalization of KLP10A and Asp, suggesting that dynein–dynactin, Asp, and KLP10A have complex interdependent functions in spindle pole focusing and centrosome attachment. These results extend recent findings from Xenopus extracts to Drosophila cultured cells and suggest that common pathways contribute to spindle pole organization and length determination.

The online version of this article contains supplemental material at MBC Online (http://www.molbiolcell.org).

Address correspondence to: Jonathan M. Scholey (jmscholey{at}ucdavis.edu).

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