Structural characterization and inhibition of the interaction between ch-TOG and TACC3
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 either 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 α-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 and delayed mitotic progression, uncovering a novel role of TACC3-ch-TOG in maintaining pericentriolar material integrity during mitosis to ensure timely cell division.
Authors
- Gabrielle Larocque (ORCID: https://orcid.org/0000-0001-8295-9378)
- James Shelford (ORCID: https://orcid.org/0000-0002-4546-9844)
- Josephina Sampson (ORCID: https://orcid.org/0000-0002-0147-6014)
- Mark A. Pfuhl (ORCID: https://orcid.org/0000-0001-9592-6639)
- Elena Rostkova
- Christian Tiede (ORCID: https://orcid.org/0000-0003-0280-4005)
- Tina Daviter (ORCID: https://orcid.org/0000-0003-2636-5959)
- Selena G. Burgess (ORCID: https://orcid.org/0000-0003-0361-0691)
- Antonio N. Calabrese (ORCID: https://orcid.org/0000-0003-2437-7761)
- Mark W. Richards (ORCID: https://orcid.org/0000-0003-1108-2825)
- Darren C. Tomlinson (ORCID: https://orcid.org/0000-0003-4134-7484)
- Richard Bayliss (ORCID: https://orcid.org/0000-0003-0604-2773)
- Stephen Royle (ORCID: https://orcid.org/0000-0001-8927-6967)
- Alistair J. Fielding
Institutions
- University of Leeds (GB)
- Institute of Cancer Research (GB)
- King's College London (GB)
- University of Warwick (GB)
- Liverpool John Moores University (GB)
Publication Details
- Journal
- The Journal of Cell Biology
- Published
- 2025-02-20
- DOI
- https://doi.org/10.1083/jcb.202407002
- Primary Topic
- Microtubule and mitosis dynamics
- Type
- article
- Field-Weighted Citation Impact
- 0.00
Funders
- Wellcome Trust
- Cancer Research UK
- Royal Society
- Directorate for Biological Sciences
- Medical Research Council
- Engineering and Physical Sciences Research Council
- Biotechnology and Biological Sciences Research Council