C-terminal Src kinase-mediated EPIYA phosphorylation of Pragmin creates a feed-forward C-terminal Src kinase activation loop that promotes cell motility.
Pragmin is one of the few mammalian proteins containing the Glu-Pro-Ile-Tyr-Ala (EPIYA) tyrosine-phosphorylation motif that was originally discovered in the Helicobacter pylori CagA oncoprotein. Following delivery into gastric epithelial cells by type IV secretion and subsequent tyrosine phosphorylation at the EPIYA motifs, CagA serves as an oncogenic scaffold/adaptor that promiscuously interacts with SH2 domain-containing mammalian proteins such as the Src homology 2 (SH2) domain-containing protein tyrosine phosphatase-2 (SHP2) and the C-terminal Src kinase (Csk), a negative regulator of Src family kinases. Like CagA, Pragmin also forms a physical complex with Csk. In the present study, we found that Pragmin directly binds to Csk by the tyrosine-phosphorylated EPIYA motif. The complex formation potentiates kinase activity of Csk, which in turn phosphorylates Pragmin on tyrosine-238 (Y238), Y343, and Y391. As Y391 of Pragmin comprises the EPIYA motif, Pragmin-Csk interaction creates a feed-forward regulatory loop of Csk activation. Together with the finding that Pragmin and Csk are colocalized to focal adhesions, these observations indicate that the Pragmin-Csk interaction, triggered by Pragmin EPIYA phosphorylation, robustly stimulates the kinase activity of Csk at focal adhesions, which direct cell-matrix adhesion that regulates cell morphology and cell motility. As a consequence, expression of Pragmin and/or Csk in epithelial cells induces an elongated cell shape with elevated cell scattering in a manner that is mutually dependent on Pragmin and Csk. Deregulation of the Pragmin-Csk axis may therefore induce aberrant cell migration that contributes to tumor invasion and metastasis.
Division of Microbiology, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan.
Yoshie
Y
Senda
Division of Microbiology, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan.
Naoko
N
Murata-Kamiya
Division of Microbiology, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan.
Masanori
M
Hatakeyama
D020816:Amino Acid Motifs; D000818:Animals; D055162:Biocatalysis; D000081247:CSK Tyrosine-Protein Kinase; D002352:Carrier Proteins / Q000737:chemistry / Q000378:metabolism*; D002465:Cell Movement; D048430:Cell Shape; D002471:Cell Transformation, Neoplastic / Q000378:metabolism* / Q000473:pathology*; D002478:Cells, Cultured; D004789:Enzyme Activation; D025461:Feedback, Physiological; D022001:Focal Adhesions; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D051379:Mice; D046912:Multiprotein Complexes / Q000737:chemistry / Q000378:metabolism; D009361:Neoplasm Invasiveness; D010766:Phosphorylation; D019000:Phosphotyrosine / Q000378:metabolism; D011485:Protein Binding; D013379:Substrate Specificity; D019061:src-Family Kinases / Q000378:metabolism*
D002352:Carrier Proteins; D047908:Intracellular Signaling Peptides and Proteins; D046912:Multiprotein Complexes; C000607534:PRAG1 protein, human; D019000:Phosphotyrosine; D000081247:CSK Tyrosine-Protein Kinase; D019061:src-Family Kinases; C000631346:CSK protein, human
C-terminal Src kinase (Csk); Cell motility; EPIYA motif; Pragmin (Sgk223); focal adhesion