Two Distinct Phosphorylation Pathways Have Additive Effects on Abl Family Kinase Activation

Abstract

The activities of the related Abl and Arg nonreceptor tyrosine kinases are kept under tight control in cells, but exposure to several different stimuli results in a two- to fivefold stimulation of kinase activity. Following the breakdown of inhibitory intramolecular interactions, Abl activation requires phosphorylation on several tyrosine residues, including a tyrosine in its activation loop. These activating phosphorylations have been proposed to occur either through autophosphorylation by Abl in trans or through phosphorylation of Abl by the Src nonreceptor tyrosine kinase. We show here that these two pathways mediate phosphorylation at distinct sites in Abl and Arg and have additive effects on Abl and Arg kinase activation. Abl and Arg autophosphorylate at several sites outside the activation loop, leading to 5.2- and 6.2-fold increases in kinase activity, respectively. We also find that the Src family kinase Hck phosphorylates the Abl and Arg activation loops, leading to an additional twofold stimulation of kinase activity. The autoactivation pathway may allow Abl family kinases to integrate or amplify cues relayed by Src family kinases from cell surface receptors.

ACKNOWLEDGMENTS

We are grateful to Xianyun Ye for technical assistance, Alex Zougman for mass spectrometric analyses, and the members of the Koleske laboratory and the three anonymous reviewers for helpful advice and comments on the manuscript.

This work was supported by NRSA grant MH67388 (K.Q.T.), USPHS grant NS39475 (A.J.K.), and a grant from the Edward J. Mallinckrodt, Jr., Foundation (A.J.K.).