Ecole Doctorale COMPLEXITE DU VIVANT – Fiche Projet CONCOURS

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Fiche à nommer selon le format Nom_Prénom_Projet18, à enregistrer en format PDF et à renvoyer à l'adresse: edcdv@upmc.fr.

Nom et prénom du directeur de thèse (et si besoin du co-directeur) : Wolff Nicolas Le directeur de thèse et le co-directeur doivent impérativement avoir l'HDR ou équivalent Coordonnées Tel : 01 45 68 88 72 e-mail : nicolas.wolff@pasteur.fr Nom et prénom du co-encadrant (non HdR ) (s’il y a lieu) : Coordonnées Tel : e-mail : Y-a-t-il un candidat déjà identifié pour le projet: OUI

Nom et prénom du responsable de l’équipe : Wolff Nicolas

Intitulé de l'équipe : Signalisation et interactions moléculaires Nombre de chercheurs et enseignants-chercheurs statutaires de l’équipe titulaires d’une HDR (ou équivalent) : 1 Nom et prénom du responsable d'unité/laboratoire ou de département: Corringer Pierre-Jean

Intitulé et N° d’unité/laboratoire ou de département: Unité Récepteurs-Canaux

Signature du directeur d'unité/laboratoire ou de département (vaut avis favorable pour le dépôt du projet) :

Dr Pierre-Jean Corringer Titre du projet de thèse : Caractérisation structurale et fonctionnelle de PDZD7, protéine impliquée dans l’audition et la vision. Spécialité : Biologie structurale, Neuroscience. Résumé du projet de thèse (1 page maximum, en anglais)

The human USHER2 syndrome, affecting the sensory cells of the inner ear and the retina, is the most common genetic cause of combined deafness and progressive blindness. Identification of the defective USHER genes has led to the characterization of major components of the hair cell protein network (Richardson et al, 2011). Many of the large modular proteins involved in these complexes have the capacity to interact with each other through several protein-protein interaction domains such as PDZ and HHD domains. Deciphering the molecular mechanisms of the hair cell protein network is an essential issue to understand both healthy and hearing associated pathologies.

Whirlin

PDZD7

+Myo7CIB2

TheUSHER2complex

Ecole Doctorale COMPLEXITE DU VIVANT – Fiche Projet CONCOURS

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Our work shall be focused on the PDZD7 protein, one of the two USHER2 scaffolding proteins along with whirlin. The PDZD7 gene is described as a genetic modifier in patients with USH2. PDZD7 has been poorly studied so far and therefore very few information are available concerning its structure and function and its interactome is mostly unknown. Modular organization of PDZD7 PDZD7 is a large multidomain protein of 1021 residues. It encompasses 4 domains of protein-protein interaction: a tandem of PDZ domains in the N-terminal region, an isolated HHD domain, and a last PDZ domain at its C-terminus. Multiple constructs of PDZD7 (mono- and multi-domains) have already been successfully cloned and expressed. We first aim at elucidating the dynamic and structural behavior of PDZD7, combining SAXS and NMR experiments. As shown for many PDZ-containing proteins, an interplay between adjacent domains could modulate the affinity and specificity of PDZD7 for ligands. PDZD7 as a key component of the USHER2 network Most Usher proteins have a PDZ binding motif at their C-terminus and are therefore potential partners for PDZD7. Associated to the USHER2 disease, two transmembrane proteins, Usherin and the G protein-coupled VLGR1, and the cytoplasmic proteins whirlin and PDZD7, form a dynamic quaternary protein complex We will first characterize the interaction between PDZD7 and the cytoplasmic regions of the two transmembrane proteins Usherin and VLGR1 encode PDZ binding motifs. Then, we will decipher the interaction between PDZD7 and its cytoplasmic homologuous whirlin. The structural heterogeneity and the dynamics of such multi-domain complexes require the use of complementary approaches combining NMR, X-ray diffraction, mass spectrometry, SAXS experiments and other biophysical methods. We have recently applied such an integrative structural approach to study the modular organization of whirlin (Delhommel et al., 2017).

These PhD tasks take place in a USHER2 project that addresses the molecular composition, structure and regulation of the USHER2 complex of the cochlear sensory hair cells during their development and functioning. It associates also structure-function of G-protein receptor (VLGR1 protein; collaboration with J.P Pin, IGF Montpellier) and ultrastructure of Usher2 complex in stereocilia using Titan Krios TEM microscope (collaboration Christine Petit and Dorit Hanein, Institut Pasteur Paris). Altogether, these biological and structural results from the macroscale (cochlea) to the microscale (protein domains) will provide important insights in how this PBM/PDZ-containing network for USHER2 complex participates in potential signal transduction as well as in hair bundle growth and morphogenesis needed for hearing.

Thèses actuellement en cours dans l’équipe Tous les encadrements doivent être indiqués (y compris les co-directions avec un autre HDR, et les encadrements dans le cadre de programmes doctoraux tels qu'IPV, FDV...)

Nom et Prénom du doctorant Directeur(s) de thèse Année de 1ère inscription

ED Financement

Genera Mariano Wolff Nicolas 2016 CdV PPU/Pasteur Zhu Yanlei Wolff Nicolas 2016 CdV ITN/Europe

PDZ PDZ PDZ HHD

Ecole Doctorale COMPLEXITE DU VIVANT – Fiche Projet CONCOURS

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Docteurs encadrés par le directeur de thèse ayant soutenu après septembre 2012 et publications relatives à leur sujet de thèse. Mettre en gras le nom du directeur de thèse et celui du docteur.

Nom Prénom : Delhommel Florent Date de soutenance : 2017 Sous la direction de Wolff Nicolas Durée de thèse (en mois): 44 Ecole Doctorale : IVIV/CdV Publications : - Bradley Worley, Florent Delhommel, Florence Cordier, Thérèse E. Malliavin, Benjamin Bardiaux , Nicolas Wolff, Michael Nilges, Carlile Lavor, Leo Liberti (2018) “Tuning interval Branch-and-Prune for protein structure determination“, J. Glob. Optim., in press.

- Delhommel F, Cordier F, Bardiaux B, Bouvier G, Colcombet-Cazenave B, Brier S, Raynal B,Nouaille S, Bahloul A, Chamot-Rooke J, Nilges M, Petit C, Wolff N (2017) “Structural characterization of whirlin reveals an unexpected and dynamic supramodule conformation of its PDZ tandem.” Structure 25(11) :1645-1656

- Delhommel F, Wolff N, Cordier F (2016) “1H, 13C and 15N backbone resonance assignments and dynamic properties of the PDZ tandem of Whirlin.” Biomol NMR Assign 10(2):361–365.

- Caillet-Saguy C, Maisonneuve P, Delhommel F, Terrien E, Babault N, Lafon M, Cordier F, Wolff N. (2015) Strategies to interfere with PDZ-mediated interactions in neurons : what we can learn from the Rabies virus. Prog Biophys Mol Biol 119 : 53-59.

Nom Prénom : Maisonneuve Pierre Date de soutenance : 2014 Sous la direction de Wolff Nicolas Durée de thèse (en mois): 44 Ecole Doctorale : IVIV/CdV Publications : - Maisonneuve P, Caillet-Saguy C, Raynal B, Gilquin B, Chaffotte A, Pérez J, Zinn-Justin S, Delepierre M, Buc H, Cordier F, Wolff N. (2014) Regulation of the catalytic activity of the human phosphatase PTPN4 by its PDZ domain. FEBS J. 281(21):4852-65.

- Caillet-Saguy C, Maisonneuve P, Delhommel F, Terrien E, Babault N, Lafon M, Cordier F, Wolff N. (2015) Strategies to interfere with PDZ-mediated interactions in neurons : what we can learn from the Rabies virus. Prog Biophys Mol Biol 119 : 53-59.

- Maisonneuve, P, Caillet-Saguy, C., Vaney, M.C., Edoo, B-Z, Sawyer, K., Raynal, B., Delepierre, M., Lafon, M., Cordier, F., Wolff, N. (2016) Molecular Basis of The Interaction of the Human Protein Tyrosine Phosphatase Non-receptor Type 4 (PTPN4) with the Mitogen-Activated Protein Kinase p38γ. J. Biol. Chem., 291(32):16699-708.

Trois publications récentes du directeur de thèse (du co-directeur ou du co-encadrant s’il y a lieu). Mettre en gras le nom du directeur de thèse.

- Delhommel,F., Cordier, F., Bardiaux, B., Bouvier, G., Colcombet-Cazenave, B., Brier, S., Raynal, B., Nouaille, S., Bahloul, A., Chamot-Rooke, J., Nilges, M., Petit, C., Wolff, N. Structural characterization of whirlin reveals an unexpected and dynamic supramodule conformation of its PDZ tandem. Structure, 2017, 25(11):1645-1656.

- Caillet-Saguy C, Toto A, Guerois R, Maisonneuve P, di Silvio E, Sawyer K, Gianni S, Wolff N. Regulation of the Human Phosphatase PTPN4 by the inter-domain linker connecting the PDZ and the phosphatase domains. Sci Rep. 2017, 7(1):7875.

- Maisonneuve P, Caillet-Saguy C., Vaney M.C., Edoo B-Z, Sawyer K., Raynal B., Delepierre M., Lafon M., Cordier F., Wolff N. Molecular Basis of The Interaction of the Human Protein Tyrosine Phosphatase Non-receptor Type 4 (PTPN4) with the Mitogen-Activated Protein Kinase p38γ. J. Biol. Chem., 2016, 291(32):16699-708.