EXCEED THE SPACE PROVIDED. We have identified a cytoskeletal associated protein kinase, called death associated protein kinase (DAPK) with a major role in apoptosis signaling. DAPK is a muti-domain ser/thr protein kinase that can phosphorylate myosin II RLC in vivo to activate acto-myosin contractile force production. In vivo DAPK is found in a complex with a discrete set of cytoskeletal, cellular and apoptosis regulatory proteins. Depletion of DAPK expression by antisense strategies induces apoptosis in cells suggesting that DAPK is an essential protein with an important function in apoptosis. We hypothesize that the proteins associated with DAPK form a cytoskeletal associated regulatory complex that functions as a 'gate-keeper' to regulate entry into the apoptotic pathway and that the cytoprotective activities of DAPK are regulated by the interaction of DAPK with this complex of proteins. One novel protein that directly binds DAPK has been identified and is called DIP (DAPK Interacting Protein). DIP has been partially characterized and shown to have 3 RING fingers with E3 ubiquitin ligase activity. Expression of DIP antagonizes the survival function of DAPK, suggesting that these apoptosis regulators have an essential role in determining a balance between survival and death. In this competitive renewal application, we propose to further test this hypothesis by examining the molecular basis through which DAPK regulates apoptosis.
Specific aim 1 is to determine the physiological role of DAPK in vascular smooth muscle cells.
Specific aim 2 is to determine the mechanism by which DAPK promotes cell survival.
Specific aim 3 is to determine the interactions and roles of the kinase domain, death domain and carboxyl-terminal tail in the regulation of apoptosis regulation by DAPK.
Aim 4 is to understand the mechanism by which DIP, antagonizes the cytoprotective function of DAPK. The proposed studies could identify a new therapeutic target that would be important in vascular diseases and will contribute to the general understanding the chain of events that occurs when cells receive death-inducing stimuli. PERFORMANCE SITE ========================================Section End===========================================
| Gallagher, Patricia J; Blue, Emily K (2014) Post-translational regulation of the cellular levels of DAPK. Apoptosis 19:306-15 |
| Zhang, Liguo; Widau, Ryan C; Herring, B Paul et al. (2011) Delta-like 1-Lysine613 regulates notch signaling. Biochim Biophys Acta 1813:2036-43 |
| Widau, Ryan C; Jin, Yijun; Dixon, Shelley A et al. (2010) Protein phosphatase 2A (PP2A) holoenzymes regulate death-associated protein kinase (DAPK) in ceramide-induced anoikis. J Biol Chem 285:13827-38 |
| Duan, Rui; Gallagher, Patricia J (2009) Dependence of myoblast fusion on a cortical actin wall and nonmuscle myosin IIA. Dev Biol 325:374-85 |
| Zhang, Liguo; Gallagher, Patricia J (2009) Mind bomb 1 regulation of cFLIP interactions. Am J Physiol Cell Physiol 297:C1275-83 |
| Zhang, Liguo; Nephew, Kenneth P; Gallagher, Patricia J (2007) Regulation of death-associated protein kinase. Stabilization by HSP90 heterocomplexes. J Biol Chem 282:11795-804 |
| Jin, Yijun; Blue, Emily K; Gallagher, Patricia J (2006) Control of death-associated protein kinase (DAPK) activity by phosphorylation and proteasomal degradation. J Biol Chem 281:39033-40 |
| Emmert, Daniel A; Fee, Judy A; Goeckeler, Zoe M et al. (2004) Rho-kinase-mediated Ca2+-independent contraction in rat embryo fibroblasts. Am J Physiol Cell Physiol 286:C8-21 |
| Jin, Yijun; Gallagher, Patricia J (2003) Antisense depletion of death-associated protein kinase promotes apoptosis. J Biol Chem 278:51587-93 |
| Blue, Emily K; Goeckeler, Zoe M; Jin, Yijun et al. (2002) 220- and 130-kDa MLCKs have distinct tissue distributions and intracellular localization patterns. Am J Physiol Cell Physiol 282:C451-60 |
Showing the most recent 10 out of 18 publications
