Use chemical inhibitors in assays examining cell division mechanisms.

A major focus of our work is to use small molecule inhibitors to turn off or turn on protein function during cell division. When combined with high-resolution microscopy analyses, these experiments can provide insight into molecular and physical mechanisms.

Questions we have answered using this approach include:

  1. How chromosomes attach to microtubules during mitosis? (Khodjakov, A. et al. JCB 2003)
  2. What are the contributions of centrosomes to bipolar spindle assembly?(Khodjakov, A. et al. JCB 2003)
  3. How do errors in chromosome-microtubule attachments arise? (Khodjakov, A. et al. JCB 2003)
  4. How are errors in chromosome-microtubule attachments corrected? (Lampson, M.A, et al. NCB 2004)
  5. How are chromosome-microtubule attachments regulated? (Lampson, M.A, et al. NCB 2005)
  6. How are different signaling pathways coordinated during cell division? (Lampson, M.A, et al. NCB 2005)

We use small molecules to inhibit proteins in cells at the lowest concentrations that have a >80% effect in a quantitative readout for the target protein’s function. Activation of protein function is achieved by exchanging the culture medium (wash-out). The kinetics of inhibition or activation are determined, and live cell experiments are designed guided by these parameters. Inhibitors with different chemical structures that target the same protein are used in parallel experiments, so that the possible overlap in any off-target effects of the compounds is minimized, particularly in the context of a narrow window of cell division biology.


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