Structural characterization and inhibition of the interaction between ch-TOG and TACC3

Shelford, J.; Burgess, S. G.; Rostkova, E.; Richards, M. W.; Tiede, C.; Fielding, A. J.; Daviter, T.; Tomlinson, D. C.; Calabrese, A. N.; Pfuhl, M.; Bayliss, R.; Royle, S. J.

doi:10.1101/2024.05.31.596836

bioRxiv · 10.1101/2024.05.31.596836

Structural characterization and inhibition of the interaction between ch-TOG and TACC3

Shelford, J.·Burgess, S. G.·Rostkova, E.·Richards, M. W.·Tiede, C.·Fielding, A. J.·Daviter, T.·Tomlinson, D. C.·Calabrese, A. N.·Pfuhl, M.·Bayliss, R.·Royle, S. J.

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Abstract

The mitotic spindle is a bipolar array of microtubules, radiating from the poles which each contain a centrosome, embedded in pericentriolar material. Two proteins, ch-TOG and TACC3, have multiple functions at the mitotic spindle due to operating alone, together or in complex with other proteins. To distinguish these activities, we need new molecular tools to dissect their function. Here, we present the structure of the -helical bundle domain of ch-TOG that mediates its interaction with TACC3 and a structural model describing the interaction, supported by biophysical and biochemical data. We have isolated Affimer tools to precisely target the ch-TOG-binding site on TACC3 in live cells, which displace ch-TOG without affecting the spindle localization of other protein complex components. Inhibition of the TACC3-ch-TOG interaction led unexpectedly to fragmentation of the pericentriolar material in metaphase cells following the formation of a bipolar spindle and delayed mitotic progression; uncovering a novel role of TACC3-ch-TOG in maintaining pericentriolar material integrity during mitosis to ensure timely cell division.

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Shelford, J., Burgess, S. G., Rostkova, E., Richards, M. W., Tiede, C., Fielding, A. J., Daviter, T., Tomlinson, D. C., Calabrese, A. N., Pfuhl, M., Bayliss, R., Royle, S. J.. 2024-05-31. Structural characterization and inhibition of the interaction between ch-TOG and TACC3. https://doi.org/10.1101/2024.05.31.596836

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