RIN1 is an ABL tyrosine kinase activator and a regulator of epithelial-cell adhesion and migration.
BACKGROUND
ABL tyrosine kinases control actin remodeling in development and in response to environmental stimuli. These changes affect cell adhesion, cell migration, and cell-cell contact. Little is known, however, about upstream mechanisms regulating ABL protein activation.


RESULTS
We report that the RAS effector RIN1 is an activator of ABL tyrosine kinases. RIN1 expression in fibroblasts promotes the formation of membrane spikes; similar effects have been reported for ABL overexpression. RIN1 binds to the ABL SH3 and SH2 domains, and these interactions stimulate ABL2 catalytic activity. This leads to increased phosphorylation of CRK and CRKL, inhibiting these cytoskeletal regulators by promoting intramolecular over intermolecular associations. Activated RAS participates in a stable RAS-RIN1-ABL2 complex and stimulates the tyrosine kinase-activation function of RIN1. Deletion of the RAS binding domain (RBD) strongly stimulated the ABL2 activation function of RIN1, suggesting that RAS activation results from the relief of RIN1 autoinhibition. The ABL binding domain of RIN1 (RIN1-ABD) increased the activity of ABL2 immune complexes and purified RIN1-ABD-stimulated ABL2 kinase activity toward CRK. Mammary epithelial cells (MECs) from Rin1-/- mice showed accelerated cell adhesion and increased motility in comparison to wild-type cells. Knockdown of RIN1 in epithelial-cell lines blocked the induction of CRKL phosphorylation, confirming that RIN1 normally functions as an inhibitor of cell motility.


CONCLUSIONS
RIN1 is a directly binding ABL tyrosine kinase activator in cells as well as in a defined-component assay. In response to environmental changes, this novel signal pathway mediates actin remodeling associated with adhesion and migration of epithelial cells.
Hu
Hailiang
H
David Geffen School of Medicine, Department of Biological Chemistry, Molecular Biology Institute, University of California, Los Angeles, Los Angeles, California 90095, USA.
Bliss
Joanne M
JM
Wang
Ying
Y
Colicelli
John
J
D000199:Actins / Q000378:metabolism; D000818:Animals; D002448:Cell Adhesion / Q000502:physiology*; D002465:Cell Movement / Q000502:physiology*; D002478:Cells, Cultured; D004789:Enzyme Activation / Q000502:physiology*; D004847:Epithelial Cells / Q000378:metabolism*; D005260:Female; D006801:Humans; D015151:Immunoblotting; D047468:Immunoprecipitation; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism*; D008321:Mammary Glands, Animal / Q000166:cytology; D051379:Mice; D016315:Proto-Oncogene Proteins c-abl / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology*; D014162:Transfection; D018909:src Homology Domains / Q000502:physiology
D000199:Actins; D047908:Intracellular Signaling Peptides and Proteins; C092117:RIN1 protein, human; D016315:Proto-Oncogene Proteins c-abl