It remains difficult to manipulate protein kinase activity with precise timing and localization in living cells. Furthermore, targeted manipulation of kinase activity only in selected protein complexes is currently impossible for the majority of biological studies. We have recently developed a new generally applicable method for rapamycin-regulated (RapR) activation of kinases and successfully applied it to three kinases from two different classes, tyr and ser/thr kinases (FAK, Src, and p38). Here, we propose to employ RapR technology to develop new broadly applicable methods for selective regulation of highly homologous kinases in living cells, and targeted activation of kinases only when they are in specific protein complexes. We will also achieve light- mediated localized regulation of kinases using caged rapamycin. These methods will be applied to identify the roles of different Src family kinases. These highly homologous kinases serve as a good test of the specificity of the new approaches and will provide new capabilities to answer previously intractable questions. Localized activation will be used to probe the spatio-temporal regulation of pathways modulating cell protrusion and polarization.

Public Health Relevance

The proposed project is focused on the development of new tools for targeted manipulation and interrogation of specific signaling pathways in live cells. In particular, the work will enable scientists to turn on kinases, an important class of regulatory molecule, in precise places and times in cells. These new approaches will allow scientists to identify biological processes critical for normal development and function of human organs as well as pathological events leading to disease.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Exploratory/Developmental Grants (R21)
Project #
1R21CA159179-01A1
Application #
8243734
Study Section
Molecular and Integrative Signal Transduction Study Section (MIST)
Program Officer
Knowlton, John R
Project Start
2012-07-01
Project End
2014-06-30
Budget Start
2012-07-01
Budget End
2013-06-30
Support Year
1
Fiscal Year
2012
Total Cost
$208,148
Indirect Cost
$77,648
Name
University of Illinois at Chicago
Department
Pharmacology
Type
Schools of Medicine
DUNS #
098987217
City
Chicago
State
IL
Country
United States
Zip Code
60612
Chu, Pei-Hsuan; Tsygankov, Denis; Berginski, Matthew E et al. (2014) Engineered kinase activation reveals unique morphodynamic phenotypes and associated trafficking for Src family isoforms. Proc Natl Acad Sci U S A 111:12420-5
Karginov, Andrei V; Tsygankov, Denis; Berginski, Matthew et al. (2014) Dissecting motility signaling through activation of specific Src-effector complexes. Nat Chem Biol 10:286-90
Dagliyan, Onur; Shirvanyants, David; Karginov, Andrei V et al. (2013) Rational design of a ligand-controlled protein conformational switch. Proc Natl Acad Sci U S A 110:6800-4