Journal article

A Combined Approach Reveals a Regulatory Mechanism Coupling Src's Kinase Activity, Localization, and Phosphotransferase-Independent Functions

Ethan Ahler, Ames C Register, Sujata Chakraborty, Linglan Fang, Emily M Dieter, Katherine A Sitko, Rama Subba Rao Vidadala, Bridget M Trevillian, Martin Golkowski, Hannah Gelman, Jason J Stephany, Alan F Rubin, Ethan A Merritt, Douglas M Fowler, Dustin J Maly

MOLECULAR CELL | CELL PRESS | Published : 2019

Abstract

Multiple layers of regulation modulate the activity and localization of protein kinases. However, many details of kinase regulation remain incompletely understood. Here, we apply saturation mutagenesis and a chemical genetic method for allosterically modulating kinase global conformation to Src kinase, providing insight into known regulatory mechanisms and revealing a previously undiscovered interaction between Src's SH4 and catalytic domains. Abrogation of this interaction increased phosphotransferase activity, promoted membrane association, and provoked phosphotransferase-independent alterations in cell morphology. Thus, Src's SH4 domain serves as an intramolecular regulator coupling catal..

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University of Melbourne Researchers

Grants

Awarded by National Institute of General Medical Sciences


Awarded by Australian National Health and Medical Research Council (NHMRC) Program Grant


Awarded by Ruth L. Kirschstein National Research Service Awards


Awarded by NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES


Funding Acknowledgements

We thank Stan Fields, Christine Queitsch, and Lea Starita for guidance. We thank Nathaniel Peters at the W.M. Keck Center for Advanced Studies in Neural Signaling for microscopy assistance and Martin Sadilek for mass spectrometry assistance. X-ray work was made possible by access to facilities at the Stanford Synchotron Radiation Lightsource supported by the Department of Energy and by the National Institutes of Health. This work was supported by the National Institute of General Medical Sciences (grant R01GM109110 to D.M.F. and grant R01GM086858 to D.J.M.). A.F.R. benefitted from an Australian National Health and Medical Research Council (NHMRC) Program Grant (1054618). The research benefitted by support from the Victorian State Government Operational Infrastructure Support and Australian Government NHMRC Independent Research Institute Infrastructure Support. D.M.F. is a CIFAR Azrieli Global Scholar. E.A. and E.M.D. were supported by a National Science Foundation Graduate Research Fellowship. E.A. and A.C.R were supported by Ruth L. Kirschstein National Research Service Awards (T32HG000035 to E.A. and T32GM008268 to A.C.R).