Crystal structure, biochemical and biophysical characterisation of NHR1 domain of E3 Ubiquitin ligase neutralized

Deepti Gupta (1) , Sylvie Beaufils (2) , Véronique Vié (2) , Gilles Paboeuf (2) , Bill Broadhurst (1) , François Schweisguth (3) , Tom L. Blundell (1) , Victor M. Bolanos-Garcia (1)

1 CSBC - Cambridge Systems Biology Centre & Department of Biochemistry
2 IPR - Institut de Physique de Rennes
3 Génétique Fonctionnelle de la Souris

Notch signaling controls diverse developmental decisions of central importance to cell activity. One of the conserved positive regulators of No- tch signaling is Neuralized, the E3 Ubiquitin li-gase enzyme that regulates signaling activity by endocytosis. Neuralized has two novel repeats, NHR1 and NHR2, with a RING finger motif at the C-terminus. Both endocytosis of the Notch ligand, Delta, and inhibition of Notch signaling by Tom, a bearded family member, require the NHR1 domain. Here we describe the first crystal structure of NHR1 domain from Drosophila me- lanogaster, solved to 2.1 Å resolution by X-ray analysis. Using NMR and other biophysical tech- niques we define a minimal binding region of Tom, consisting of 12 residues, which interacts with NHR1 and show by interfacial analysis of protein monolayers that NHR1 binds PI4P. Taken together, the studies provide insight into mo-lecular interactions that are important for Notch signaling.

Mots clés: Notch; Signaling; Neuralized; NHR1; Bearded; Tom; Endocytosis; Neurogenesis

Domaines: Matière Molle [cond-mat.soft]

Deepti Gupta, Sylvie Beaufils, Véronique Vié, Gilles Paboeuf, Bill Broadhurst, et al.. Crystal structure, biochemical and biophysical characterisation of NHR1 domain of E3 Ubiquitin ligase neutralized. Advances in enzyme research, 2013, 1 (3), pp.61-75. ⟨10.4236/aer.2013.13007⟩. ⟨hal-00939236⟩