The SH3 regulatory domain of the hematopoietic cell kinase Hck binds ELMO via its polyproline motif

FEBS Open Bio. 2015 Feb 4;5:99-106. doi: 10.1016/j.fob.2015.01.009. eCollection 2015.


Eukaryotic EnguLfment and cell MOtility (ELMO) proteins form an evolutionary conserved family of regulators involved in small GTPase dependent actin remodeling processes that regulates the guanine exchange factor activity of some of the Downstream Of CrK (DOCK) family members. Gathered data strongly suggest that DOCK activation by ELMO and the subsequent signaling result from a subtle balance in the binding of partners to ELMO. Among its putative upward modulators, the Hematopoietic cell kinase (Hck), a member of the Src kinase superfamily, has been identified as a binding partner and a specific tyrosine kinase for ELMO1. Indeed, Hck is implicated in distinct molecular signaling pathways governing phagocytosis, cell adhesion, and migration of hematopoietic cells. Although ELMO1 has been shown to interact with the regulatory Src Homology 3 (SH3) domain of Hck, no direct evidence indicating the mode of interaction between Hck and ELMO1 have been provided in the literature. In the present study, we report convergent pieces of evidence that demonstrate the specific interaction between the SH3 domain of Hck and the polyproline motif of ELMO1. Our results also suggest that the tyrosine-phosphorylation state of ELMO1 tail might act as a putative modulator of Hck kinase activity towards ELMO1 that in turn participates in DOCK180 activation and further triggers subsequent signaling towards actin remodeling.

Keywords: DOCK, Downstream Of CrK protein family; EAD, ELMO Autoregulatory Domain; EID, ELMO Inhibitory Domain; ELMO; ELMO, EnguLfment and cell MOtility protein family; ERM, Ezrin–Radixin–Moesin protein family; FRET, Förster (Fluorescence) resonance energy transfer; GEF, Guanine nucleotide Exchange Factor; GSH, Glutathione (reduced); GST, Glutathione S-Transferase; Hck; Hck, Hematopoietic cell kinase; PH, Pleckstrin Homology domain; Phagocytosis; Phosphorylation; Polyproline; PxP, Polyproline motif; RBD, Rho-Binding Domain; SH3; SH3, Src Homology 3 domain; TAMs, Tyro3, Axl and Mer receptor tyrosine kinase family; TEV, Tobacco Etch Virus.