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Open Access Research

Inhibition of clathrin by pitstop 2 activates the spindle assembly checkpoint and induces cell death in dividing HeLa cancer cells

Charlotte M Smith1, Volker Haucke23, Adam McCluskey4, Phillip J Robinson1 and Megan Chircop15*

Author Affiliations

1 Children's Medical Research Institute, The University of Sydney, 214 Hawkesbury Road, Westmead, NSW, 2145, Australia

2 Institute of Chemistry and Biochemistry & Neurocure Cluster of Excellence, Freie Universität Berlin, Berlin, 14195, Germany

3 Leibniz-Institut für Molekulare Pharmakologie (FMP), Berlin-Buch, Germany

4 Chemistry, School of Environmental & Life Sciences, The University of Newcastle, Callaghan, NSW, 2308, Australia

5 Children’s Medical Research Institute, Locked Bag 23, Wentworthville, NSW, 2145, Australia

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Molecular Cancer 2013, 12:4  doi:10.1186/1476-4598-12-4

Published: 17 January 2013

Abstract

Background

During metaphase clathrin stabilises the mitotic spindle kinetochore(K)-fibres. Many anti-mitotic compounds target microtubule dynamics. Pitstop 2™ is the first small molecule inhibitor of clathrin terminal domain and inhibits clathrin-mediated endocytosis. We investigated its effects on a second function for clathrin in mitosis.

Results

Pitstop 2 did not impair clathrin recruitment to the spindle but disrupted its function once stationed there. Pitstop 2 trapped HeLa cells in metaphase through loss of mitotic spindle integrity and activation of the spindle assembly checkpoint, phenocopying clathrin depletion and aurora A kinase inhibition.

Conclusions

Pitstop 2 is therefore a new tool for investigating clathrin spindle dynamics. Pitstop 2 reduced viability in dividing HeLa cells, without affecting dividing non-cancerous NIH3T3 cells, suggesting that clathrin is a possible novel anti-mitotic drug target.

Keywords:
Clathrin; Pitstop; Spindle assembly checkpoint; Metaphase; Cell death; Cancer