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Journal Articles Molecular and Cellular Proteomics Year : 2023

SKAP2 Modular Organization Differently Recognizes SRC Kinases Depending on Their Activation Status and Localization

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Abstract

Dimerization of SRC kinase adaptor phosphoprotein 2 (SKAP2) induces an increase of binding for most SRC kinases suggesting a fine-tuning with transphosphorylation for kinase activation. This work addresses the molecular basis of SKAP2-mediated SRC kinase regulation through the lens of their interaction capacities. By combining a luciferase complementation assay and extensive site-directed mutagenesis, we demonstrated that SKAP2 interacts with SRC kinases through a modular organization depending both on their phosphorylation-dependent activation and subcellular localization. SKAP2 contains three interacting modules consisting in the dimerization domain, the SRC homology 3 (SH3) domain, and the second interdomain located between the Pleckstrin homology and the SH3 domains. Functionally, the dimerization domain is necessary and sufficient to bind to most activated and myristyl SRC kinases. In contrast, the three modules are necessary to bind SRC kinases at their steady state. The Pleckstrin homology and SH3 domains of SKAP2 as well as tyrosines located in the interdomains modulate these interactions. Analysis of mutants of the SRC kinase family member hematopoietic cell kinase supports this model and shows the role of two residues, Y390 and K7, on its degradation following activation. In this article, we show that a modular architecture of SKAP2 drives its interaction with SRC kinases, with the binding capacity of each module depending on both their localization and phosphorylation state activation. This work opens new perspectives on the molecular mechanisms of SRC kinases activation, which could have significant therapeutic impact.
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Origin : Publication funded by an institution
Licence : CC BY - Attribution
Licence : CC BY - Attribution
Licence : CC BY NC - Attribution - NonCommercial

Dates and versions

pasteur-03953675 , version 1 (24-01-2023)

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Attribution - CC BY 4.0

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Laurine Levillayer, Patricia Cassonnet, Marion Declercq, Mélanie Dos Santos, Louis Lebreton, et al.. SKAP2 Modular Organization Differently Recognizes SRC Kinases Depending on Their Activation Status and Localization. Molecular and Cellular Proteomics, 2023, 22 (1), pp.100451. ⟨10.1016/j.mcpro.2022.100451⟩. ⟨pasteur-03953675⟩
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