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Grenoble Institut des Neurosciences Grenoble Institut des Neurosciences

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Study of the APC-EB1/3-Microtubule complex by biochemistry and electron microscopy

Objectifs

Our objectives are to characterize the ternary complex between APC, EB1/3 and the microtubules by biochemical methods, video microscopy and electron microscopy.

Résumé

Discovered for its direct link with colon cancer, the Adenomatous Polyposis Coli protein (APC) was first characterized for preventing the transcription of specific oncogenes. In neurons, APC is involved in the development of axon and post-synaptic compartments and APC mutations are linked to neurologic disabilities related to autism or schizophrenia. These functions are connected to the cytoskeleton interacting properties of APC. In this context, APC is associated to the end-binding proteins (EBs). EBs form a complex interaction network at microtubule plus-ends, where they control microtubule dynamics and microtubule anchorage to distinct subcellular targets (e.g. cell cortex, kinetochores). By video microscopy, we showed that APC enhanced EB1/3 binding to microtubule ends. We would like to understand this property at the “atomic level” by studying the APC-EB1/3-microtubule complex by biochemistry, video and electron microscopy.

Méthodes

Protein purification - interaction assay - fluorescence microscopy - negative staining electron microscopy - cryo-electron microscopy

Références

  • Ramirez-Rios S, Denarier E, Prezel E, Vinit A, Stoppin-Mellet V, Devred F, Barbier P, Peyrot V, Sayas CL, Avila J, Peris L, Andrieux A, Serre L, Fourest-Lieuvin A, Arnal I. (2016) Tau antagonizes end-binding protein tracking at microtubule ends through a phosphorylation-dependent mechanism. Mol. Cell. Biol. 27, 2924-34.
  • Velot L, Molina A, Rodrigues-Ferreira S, Nehlig A, Bouchet BP, Morel M, Leconte L, Serre L, Arnal I, Braguer D, Savina A, Honore S and Nahmias C (2015) Negative regulation of EB1 turnover at microtubule plus ends by interaction with microtubule-associated protein ATIP3. Oncotarget 6, 43557-70
  • Elie A, Prezel E, Guérin C, Denarier E, Ramirez-Rios S, Serre L, Andrieux A, Fourest-Lieuvin A, Blanchoin L and Arnal I (2015) Tau co-organizes dynamic microtubule and actin networks. Scientific Reports, 5:9964. doi: 10.1038/srep09964

Contacts

L. Serre, CR1-CNRS
Email : laurence.serre@univ-grenoble-alpes.fr

Mise à jour le 23 mai 2017

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