Abstract
The regulation of mitotic spindle orientation is essential to ensure proper cell division and development (Kiyomitsua and Cheeseman Nat Cell Biol 14:311–317, 2012). For identification of potential spindle orientation regulators, determination of the mitotic spindle angle is a well-known but time-consuming procedure. Here we describe a simple and time-saving phenotypic screening assay for the identification of potential spindle orientation regulators. This screen is based on the analysis of monopolar mitotic spindle structures, which form upon inhibition of the mitotic kinesin Eg5/KSP by the small-molecule inhibitor dimethylenastron (DME) or similar compounds.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Kiyomitsu T, Cheeseman IM (2012) Chromosome- and spindle-pole-derived signals generate an intrinsic code for spindle position and orientation. Nat Cell Biol 14:311–317
Pease JC, Tirnauer JS (2011) Mitotic spindle misorientation in cancer – out of alignment and into the fire. J Cell Sci 124:1007–1016
Bergstralh DT, St Johnston D (2014) Spindle orientation: what if it goes wrong? Semin Cell Dev Biol 34:140–145
Ertych N, Stolz A, Stenzinger A, Weichert W, Kaulfuß S, Burfeind P et al (2014) Increased microtubule assembly rates mediate chromosomal instability in colorectal cancer cells. Nat Cell Biol 16:779–791
Stolz A, Ertych N, Bastians H (2015) A phenotypic screen identifies microtubule plus end assembly regulators that can function in mitotic spindle orientation. Cell Cycle 14:827–837
Kapoor TM, Mayer TU, Coughlin ML, Mitchison TJ (2000) Probing spindle assembly mechanisms with monastrol, a small molecule inhibitor of the mitotic kinesin, Eg5. J Cell Biol 150:975–988
Mayer TU, Kapoor TM, Haggarty SJ, King RW, Schreiber SL, Mitchison TJ (1999) Small molecule inhibitor of mitotic spindle bipolarity identified in phenotype-based screen. Science 286:971–974
Müller C, Gross D, Sarli V, Gartner M, Giannis A, Bernhardt G et al (2007) Inhibitors of kinesin Eg5: antiproliferative activity of monastrol analogues against human glioblastoma cells. Cancer Chemother Pharmacol 59:157–164
Acknowledgments
We are grateful to the Deutsche Forschungsgemeinschaft and the German Cancer Aid for financial support.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2018 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Glaubke, E., Bastians, H. (2018). A Cell-Based Assay for Mitotic Spindle Orientation. In: Wagner, B. (eds) Phenotypic Screening. Methods in Molecular Biology, vol 1787. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7847-2_5
Download citation
DOI: https://doi.org/10.1007/978-1-4939-7847-2_5
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7846-5
Online ISBN: 978-1-4939-7847-2
eBook Packages: Springer Protocols