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Grenoble Institut des Neurosciences Grenoble Institut des Neurosciences

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Team "Cytoskeleton dynamics and structure"

Director: Isabelle ARNAL

Study the molecular mechanisms of microtubule regulation by cytoskeletal-associated proteins, using complementary approaches of biochemistry, optical video-microscopy and electron cryomicroscopy.
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The team "Cytoskeleton dynamics and structure" is known for its expertise in the analysis of the architecture of the cytoskeleton of the nanoscale to the three-dimensional molecular organization.

Research themes and techniques used

Microtubules are major components of the eukaryotic cytoskeleton and are involved in cell division, motility and morphogenesis. Important for these functions are the dynamic and structural properties of microtubules, which are finely regulated in vivo by numerous microtubule-associated proteins (MAPs).

Our team focuses on the molecular basis underlying microtubule regulation by MAPs that are specifically involved in cytoskeleton organisation in mitosis, neuronal differentiation and neuronal plasticity. For that, we use light microscopy imaging to study microtubule properties in both simple cell-free systems and cellular differentiation models. In parallel, we are using electron cryomicroscopy and cryotomography methods to investigate how the MAPs of interest precisely interact with microtubules and affect the structure and spatial arrangement of these polymers.

This correlative approach combining light- and electron microscopy on various complexity level systems should help to understand at a molecular level how a MAP network elaborate complex cytoskeleton architecture in cells.

Techniques used:

  • Biochemistry and cellular biology: subcloning, recombinant protein expression, protein purification, centrosome purification, preparation of cellular extracts, study of protein interactions, spectrophotometry, stable cell line and neuronal primary cell cultures.

  • Light microscopy: total internal reflection fluorescence microscopy, video fluorescence microscopy, confocal microscopy.

  • Electron microscopy: negative stain, electron cryomicroscopy and cryotomography.
 

 See our publications on TIRF biomimetic assays to reconstitute EB binding at MT ends or microtubules and actin co-assembly.
 

 

Publications

Serre L, Stoppin-Mellet V and Arnal I (2019) Adenomatous Polyposis Coli as a scaffold for microtubule End-Binding proteins J Mol Biol 43: 1993

Peris L, Bisbal M, Martinez-Hernandez J, Saoudi Y, Jonckheere J, Rolland M, Sebastien M, Brocard J, Denarier E, Bosc C, Guerin C, Gory-Faure S, Deloulme JC, Lante F, Arnal I, Buisson A, Goldberg Y, Blanchoin L, Delphin C and Andrieux A (2018) A key function for microtubule-associated-protein 6 in activity-dependent stabilisation of actin filaments in dendritic spines Nat Commun 9: 3775

Prezel E, Elie A, Delaroche J, Stoppin-Mellet V, Bosc C, Serre L, Fourest-Lieuvin A, Andrieux A, Vantard M and Arnal I (2018) Tau can switch microtubule network organizations: from random networks to dynamic and stable bundles Mol Biol Cell 29: 154

Prezel E, Stoppin-Mellet V, Elie A, Zala N, Denarier E, Serre L and Arnal I (2017) TIRF assays for real-time observation of microtubules and actin coassembly: Deciphering tau effects on microtubule/actin interplay Methods Cell Biol 141: 199

Ramirez-Rios S, Serre L, Stoppin-Mellet V, Elie E, Vinit A, Courriol E, Fourest-Lieuvin A, Delaroche J, Denarier E and Arnal I (2017) A TIRF microscopy assay to decode how tau regulates EB's tracking at microtubule ends Methods Cell Biol 141: 179

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 and Arnal I (2016) Tau antagonizes end-binding protein tracking at microtubule ends through a phosphorylation-dependent mechanism Mol Biol Cell 27: 2924

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. Sci Rep 5:9964. doi: 10.1038/srep09964

Sayas CL, Tortosa E, Bollati F, Ramirez-Rios S, Arnal I and Avila J (2015) Tau regulates the localization and function of End binding proteins 1 and 3 (EB1/3) in neuronal cells. J Neurochem 133, 653-667

Delphin C., Bouvier D., Seggio M., Couriol E., Saoudi Y., Denarier E., Bosc C., Valiron O., Bisbal M., Arnal I. and Andrieux A. (2012). MAP6-F is a temperature sensor that directly binds to and protects microtubules from cold-induced depolymerization. J. Biol. Chem., 287, 35127-35138

Updated on November 12, 2019

Fundings

 
 

Collaborators

L. Blanchoin (CEA, Grenoble)
G. Schoen (IBS, Grenoble)
F. Forbes (INRIA, Grenoble)
I. Kulic (ICS, Strasbourg)
H. Mohrbach (LPCT, Nancy)
E. Lebarbier (AgroParisTech)
N. Galjart (Rotterdam)

Internship

Anthony Manon (Master) 2020
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