Regulation of microtubule polymerization and depolymerization is required for proper cell development. Here, we report that two proteins of the Drosophila melanogaster kinesin-13 family, KLP10A and KLP59C, cooperate to drive microtubule depolymerization in interphase cells. Analyses of microtubule dynamics in S2 cells depleted of these proteins indicate that both proteins stimulate depolymerization, but alter distinct parameters of dynamic instability; KLP10A stimulates catastrophe (a switch from growth to shrinkage) whereas KLP59C suppresses rescue (a switch from shrinkage to growth). Moreover, immunofluorescence and live analyses of cells expressing tagged kinesins reveal that KLP10A and KLP59C target to polymerizing and depolymerizing microtubule plus ends, respectively. Our data also suggest that KLP10A is deposited on microtubules by the plus-end tracking protein, EB1. Our findings support a model in which these two members of the kinesin-13 family divide the labour of microtubule depolymerization.
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We thank P. Sampaio and C. Sunkel (Porto, Portugal) for the anti-Mast antibody, E. Ghersi and L. D'adamio (AECOM) for advice on in vitro binding assays, M. Cammer of the AECOM analytical imaging facility for advice on image acquisition and analysis, and H. Sosa (AECOM) for helpful comments on the manuscript. We also thank R. Tsien (UCSD) for providing the mRFP construct, and K. Slep (UCSF) for the APC and some EB1 constructs. This work was supported by grants from the NIH to D.J.S. and R.D.V. V.M. is a Fulbright Fellow and D.J.S. is a Scholar of the Leukemia and Lymphoma Society.
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