Research in my laboratory focuses on the interrogation and manipulation of signaling pathways regulating cell motility and cell adhesion. We particularlyemphasize our studies on events regulated by protein phosphorylation. By controlling phosphorylation of specific targets, protein kinases and phosphatases regulate signaling pathways thatdetermine morphology and migratory behavior of a cell. These events control a multitude of physiological and pathological processes during development and normal functioning of a multicellular organism. Detailed understanding of these signaling networks will not only provide us knowledge of how specific organs work, but also help us to identify pathological markers and therapeutic targets.
To address the most pressing questions we develop innovative technologies that allow us to overcome limitations of existing methods and tackle scientific problems from a different perspective. One of the directions in my laboratory is based on a novel method for regulation of kinase activity in living cells. Using a combination of an engineered allosteric switch, the iFKBP domain, and a small membrane permeable molecule, rapamycin or its non-immunosuppresive analogs, we can now control the activity of a selected kinase with high specificity and precise temporal control. Using this method we have already developed rapamycin-regulated (RapR) analogs of several kinases, representing both Tyr and Ser/Thr kinases (FAK, Src, Fyn, Yes, Pak1 and p38). Currently we are working on extending this technology to manipulate specific pathways regulated by a selected kinase.
Most studies of kinase function are focused on either the identification of events affected by kinase inactivation (i.e. use of pharmacological inhibitors) or studying the effect of sustained kinase activity (i.e. overexpression of active kinase). However, activation of a kinase often triggers a complex multistage cascade of biochemical and morphological changes in the cell. More detailed studies are required to fully understand these processes. Thus, our research focuses on the identification of events triggered by a specific kinase at different time points after its activation.By manipulating a specific kinase or a particular pathway we are trying to dissect individual processes involved in regulation of cell adhesion and migration. Application of the latest methods for live cell imaging in combination with novel approaches for image analysis allows us to determine, quantify and correlate spatiotemporal changes in cell morphology, subcellular structures, and the activity of specific proteins.