14-3-3 proteins inactivate DAPK2 by promoting its dimerization and protecting key regulatory phosphosites

0545402 - FGÚ 2022 RIV GB eng J - Journal Article
Horváth, Matej - Petrvalská, Olivia - Herman, P. - Obšilová, Veronika - Obšil, Tomáš
14-3-3 proteins inactivate DAPK2 by promoting its dimerization and protecting key regulatory phosphosites.
Communications Biology. Roč. 4, č. 1 (2021), č. článku 986. E-ISSN 2399-3642
R&D Projects: GA ČR(CZ) GA19-00121S
Research Infrastructure: CIISB II - 90127
Institutional support: RVO:67985823
Keywords : protein structure and function * protein kinase * DAPK * 14-3-3 protein * apoptosis * phosphorylation
OECD category: Biochemistry and molecular biology
Impact factor: 6.548, year: 2021
Method of publishing: Open access
https://doi.org/10.1038/s42003-021-02518-y

Death-associated protein kinase 2 (DAPK2) is a CaM-regulated Ser/Thr protein kinase, involved in apoptosis, autophagy, granulocyte differentiation and motility regulation, whose activity is controlled by autoinhibition, autophosphorylation, dimerization and interaction with scaffolding proteins 14-3-3. However, the structural basis of 14-3-3-mediated DAPK2 regulation remains unclear. Here, we structurally and biochemically characterize the full-length human DAPK2:14-3-3 complex by combining several biophysical techniques. The results from our X-ray crystallographic analysis revealed that Thr369 phosphorylation at the DAPK2 C terminus creates a high-affinity canonical mode III 14-3-3-binding motif, further enhanced by the diterpene glycoside Fusicoccin A. Moreover, concentration-dependent DAPK2 dimerization is disrupted by Ca2+/CaM binding and stabilized by 14-3-3 binding in solution, thereby protecting the DAPK2 inhibitory autophosphorylation site Ser318 against dephosphorylation and preventing Ca2+/CaM binding. Overall, our findings provide mechanistic insights into 14-3-3-mediated DAPK2 inhibition and highlight the potential of the DAPK2:14-3-3 complex as a target for anti‐inflammatory therapies.
Permanent Link: http://hdl.handle.net/11104/0322100