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The Journal of Immunology, 2003, 171: 519-523.
Copyright © 2003 by The American Association of Immunologists

CUTTING EDGE

Cutting Edge: A Chemical Genetic System for the Analysis of Kinases Regulating T Cell Development 1

Angela Denzel*, Katherine J. Hare{dagger}, Chao Zhang{ddagger}, Kevan Shokat{ddagger}, Eric J. Jenkinson{dagger}, Graham Anderson2,{dagger} and Adrian Hayday2,3,*

* Department of Immunobiology, New Guy’s House, Guy’s, King’s and St. Thomas’s School of Medicine, London, United Kingdom; {dagger} Department Anatomy, Medical Research Council Center for Immune Regulation University of Birmingham, Birmingham, United Kingdom; and {ddagger} Department of Cellular and Molecular Pharmacology, University of California San Francisco, CA 94143

To understand the regulatory activities of kinases in vivo requires their study across a biologically relevant window of activity. To this end, ATP analog-sensitive kinase alleles (ASKAs) specifically sensitive to a competitive inhibitor have been developed. This article tests whether ASKA technology can be applied to complex immunological systems, such as lymphoid development. The results show that when applied to reaggregate thymic organ culture, novel p56Lck ASKAs readily expose a dose-dependent correlation of thymocyte development with a range of p56Lck activity. By regulating kinase activity, rather than amounts of RNA or protein, ASKA technology offers a general means for assessing the quantitative contributions to immunology of numerous kinases emerging from genomics analyses. It can obviate the generation of multiple lines of mice expressing different levels of kinase transgenes and should permit specific biological effects to be associated with defined biochemical activities.




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