Biophysical Studies of Mitotic Microtubule Dynamics and Spindle Function

Our research group uses a combination experimental and computational approach to dissect molecular mechanisms for how cells divide, and for how cell division can be controlled to prevent genetic diseases, pathogenic anti-fungal drug resistance, and cancer. Specifically, we are interested in how proteins regulate the dynamics and mechanics of critical cell division components. These components include microtubules, which align duplicated chromosomes and then pull them apart during cell division, and also the chromosomes themselves, whose own mechanics contribute to their proper segregation during mitosis. Our overarching goal is to improve human health by developing an improved mechanistic understanding for how cells divide.

The primary experimental tools in the Gardner lab are single-molecule Total Internal Reflection Fluorescence (TIRF) microscopy, and electron microscopy. We image living cells using quantitative fluorescence microscopy, and also purify proteins such as tubulin (which is the building block of microtubules) and other microtubule-associated proteins to observe their interactions outside of the cell. We use biophysical computational modeling to better integrate and understand our experimental observations, make new experimental predictions, and to test whether our proposed cellular mechanisms are physically reasonable. We broaden all of our experimental observations with extensive quantitative image analysis, in order to squeeze as much information as possible out of every collected experimental image. Overall, we are a cellular biophysics laboratory that combines cell biology tools with biophysical methods to shed new light on the regulation of cell division.

Reid TA, Coombes C, Mukherjee S, Goldblum RR, White K, Parmar S, McClellan M, Zanic M, Courtemanche N, Gardner MK. (2019) Structural state recognition facilitates tip tracking of EB1 at growing microtubule ends. Elife. 2019 Sep 3;8. pii: e48117. doi: 10.7554/eLife.48117.

Centromere mechanical maturation during mammalian cell mitosis. Harasymiw LA, Tank D, McClellan M, Panigrahy N, Gardner MK. Nat Commun. 2019 Apr 15;10(1):1761. doi: 10.1038/s41467-019-09578-z.

A Gradient in Metaphase Tension Leads to a Scaled Cellular Response in Mitosis. Mukherjee S, Sandri BJ, Tank D, McClellan M, Harasymiw LA, Yang Q, Parker LL, Gardner MK.Dev Cell. 2019 Apr 8;49(1):63-76.e10. doi: 10.1016/j.devcel.2019.01.018. Epub 2019 Feb 21.

Reid TA, Coombes C, Gardner MK., Manipulation and quantification of microtubule lattice integrity., Biology open. 2017; 6(8):1245-1256.

Coombes C, Yamamoto A, McClellan M, Reid TA, Plooster M, Luxton GW, Alper J, Howard J, Gardner MK.; Mechanism of microtubule lumen entry for the α-tubulin acetyltransferase enzyme αTAT1., Proc Natl Acad Sci U S A. 2016 Nov 15;113(46):E7176-E7184.

TA Reid, BM Schuster, BJ Mann, SK Balchand, M Plooster, M McClellan, CE Coombes, P Wadsworth, and MK Gardner, “Suppression of Microtubule Assembly Kinetics by the Mitotic Protein TPX2” Journal of Cell Science (2015)

Hepperla A.J., Willey P.T., Coombes C.E., Schuster B.M., Gerami-Nejad M., McClellan M., Mukherjee S., Meehl J.B., Winey M., Odde D.J., O'Toole E., Gardner M.K., “Minus-end-directed Kinesin-14 motors align antiparallel microtubules to control metaphase spindle length”, Developmental Cell; 31(1):61-72 (2014).

Chacón, J.M., Mukherjee, S., Schuster, B.M., Clarke, D.J., and Gardner, M.K., “Pericentromere Tension is Self-Regulated by Spindle Structure in Metaphase”, Journal of Cell Biology; 205(3):313-24 (2014).

Chacón, J.M. and Gardner, M.K., “Analysis and Modeling of Chromosome Congression During Mitosis in the Chemotherapy Drug Cisplatin”, Cellular and Molecular Bioengineering; 6(4): 406-17(2013).

Coombes, C.E., Yamamoto A., Kenzie, M.R., Odde, D.J., Gardner M.K., “Evolving tip structures can explain age-dependent microtubule catastrophe”, Curr Biol. Jul 22; 23(14):1342-8 (2013).

Gardner, M.K., Zanic, M., Gell, C., Bormuth, V., and J. Howard, “Depolymerizing Kinesins Kip3 and MCAK Shape Cellular Microtubule Architecture by Differential Control of Catastrophe”, Cell 147(5):1092-103 (2011)

Gardner, M.K., Charlebois, B.D., Janosi, I.M., Howard, J., Hunt, A.J., and D.J. Odde, “Rapid Microtubule Self-assembly Kinetics”, Cell 146(4):1247-60. (2011)