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Expression Levels of a Kinesin-13 Microtubule Depolymerase Modulates the Effectiveness of Anti-Microtubule Agents

Title: Expression Levels of a Kinesin-13 Microtubule Depolymerase Modulates the Effectiveness of Anti-Microtubule Agents
Authors: Schimizzi, Gregory V.; Currie, Joshua D.; Rogers, Stephen L.
Source: PloS One, 5(6)
Publication Year: 2010
Collection: Carolina Digital Repository (UNC - University of North Carolina)
Subject Terms: Microscopy; Kinesin; Science; Cell Biology/Cytoskeleton; Cell Biology/Cell Growth and Division; Fluorescence; DNA Primers; Microtubules; Animals; Cultured; Biochemistry/Chemical Biology of the Cell; Research Article; Drosophila; Base Sequence; RNA Interference; Cells; Medicine
Description: BackgroundChemotheraputic drugs often target the microtubule cytoskeleton as a means to disrupt cancer cell mitosis and proliferation. Anti-microtubule drugs inhibit microtubule dynamics, thereby triggering apoptosis when dividing cells activate the mitotic checkpoint. Microtubule dynamics are regulated by microtubule-associated proteins (MAPs); however, we lack a comprehensive understanding about how anti-microtubule agents functionally interact with MAPs. In this report, we test the hypothesis that the cellular levels of microtubule depolymerases, in this case kinesin-13 s, modulate the effectiveness of the microtubule disrupting drug colchicine.Methodology/Principal FindingsWe used a combination of RNA interference (RNAi), high-throughput microscopy, and time-lapse video microscopy in Drosophila S2 cells to identify a specific MAP, kinesin-like protein 10A (KLP10A), that contributes to the efficacy of the anti-microtubule drug colchicine. KLP10A is an essential microtubule depolymerase throughout the cell cycle. We find that depletion of KLP10A in S2 cells confers resistance to colchicine-induced microtubule depolymerization to a much greater extent than depletion of several other destabilizing MAPs. Using image-based assays, we determined that control cells retained 58% (±2%SEM) of microtubule polymer when after treatment with 2 µM colchicine for 1 hour, while cells depleted of KLP10A by RNAi retained 74% (±1%SEM). Likewise, overexpression of KLP10A-GFP results in increased susceptibility to microtubule depolymerization by colchicine.Conclusions/SignificanceOur results demonstrate that the efficacy of microtubule destabilization by a pharmacological agent is dependent upon the cellular expression of a microtubule depolymerase. These findings suggest that expression levels of Kif2A, the human kinesin-13 family member, may be an attractive biomarker to assess the effectiveness of anti-microtubule chemotherapies. Knowledge of how MAP expression levels affect the action of anti-microtubule drugs may prove useful ...
Document Type: article in journal/newspaper
Language: English
Relation: https://cdr.lib.unc.edu/downloads/sn00b529t?file=thumbnail; https://cdr.lib.unc.edu/downloads/sn00b529t
DOI: 10.17615/1e34-ef14
Availability: https://doi.org/10.17615/1e34-ef14; https://cdr.lib.unc.edu/downloads/sn00b529t?file=thumbnail; https://cdr.lib.unc.edu/downloads/sn00b529t
Rights: http://rightsstatements.org/vocab/InC/1.0/
Accession Number: edsbas.4E784207
Database: BASE