14-3-3-protein regulates Nedd4-2 by modulating interactions between HECT and WW domains

P. Pohl1, T. Obsil1,2 a V. Obsilova1

1Dept. of Structural Biology of Signalling Proteins, Division BIOCEV, Institute of Physiology of the Czech Academy of Sciences, Vestec, Czech Republic

2Dept. of Physical and Macromolecular Chemistry, Faculty of Science, Charles University in Prague, Czech Republic

pavel.pohl@fgu.cas.cz

 

Neural precursor cell expressed developmentally down-regulated 4 ligase (Nedd4-2) is an E3 ubiquitin ligase that targets proteins for ubiquitination and endocytosis, thereby regulating numerous ion channels, membrane receptors and tumor suppressors. Nedd4-2 activity is regulated by autoinhibition, calcium binding, oxidative stress, substrate binding, phosphorylation and 14-3-3 protein binding [1-3]. However, the structural basis of 14-3-3-mediated Nedd4-2 regulation remains poorly understood. Here, we combined several techniques of integrative structural biology to characterize Nedd4-2 and its complex with 14-3-3. We demonstrate that phosphorylated Ser342 and Ser448 are the key residues that facilitate 14-3-3 protein binding to Nedd4-2 and that 14-3-3 protein binding induces a structural rearrangement of Nedd4-2 by inhibiting interactions between its structured domains. Overall, our findings provide the structural glimpse into the 14-3-3-mediated Nedd4-2 regulation and highlight the potential of the Nedd4-2:14-3-3 complex as a pharmacological target for Nedd4-2-associated diseases such as hypertension, epilepsy, kidney disease and cancer [4-5].

1. P. Goel, J. A. Manning, and S. Kumar, Gene, 557, (2015), 110.

2. H. He, C. Huang, Z. Chen, H. Huang, X. Wang and J. Chen, Biomed Pharmacother, 125, (2020), 109983.

3. J. A. Manning and S. Kumar, Trends Biochem. Sci., 43, (2018), 635647.

4. P. Pohl, R Joshi, O. Petrvalska, T. Obsil, V. Obsilova. Commun Biol., 4, (2021), 899.

5. R. Joshi, P. Pohl, D. Strachotova, P. Herman, T. Obsil, V. Obsilova. Biophys. J., accepted (2022), DOI: https://doi.org/10.1016/j.bpj.2022.02.025

 

This study was supported by the Czech Science Foundation (Project 20-00058S) and the Czech Academy of Sciences (Research Projects RVO: 67985823 of the Institute of Physiology).