The long term goal of our research is to understand the molecular mechanisms driving the propagation and termination of protein kinase C (PKC) signaling in response to agonists in cells. PKC has been in the spotlight since the discovery three decades ago that it is activated by the lipid second messenger, diacylglycerol. Despite PKC's enduring stage presence and tremendous advances in understanding the enzymology and regulation of this key protein, understanding the function of PKC in biology is still under intense pursuit. We have previously focused on understanding the molecular mechanisms of PKC as a first step in understanding how this key protein functions in the cell. Under the auspices of the MERIT Award, we ventured away from our biochemical studies and dove into the cell, asking the guestion: what does protein kinase C do in the cell? We developed novel genetically-encoded reporters that allowed us to simultaneously visualize PKC activity and second messenger production in real time in live cells. In the next funding period, we propose to take this new technology to the next level and develop reporters that will allow us to simultaneously image two or more kinase activities at in the same cell. This will allow us to examine the interplay between different signaling pathways, feedback mechanisms, and identify regulatory inputs that our previous biochemical studies could not address. The three major questions we ask are: 1] how does PKC impact other signaling pathways? 2] how do phosphatases control signal amplitude and signal termination? and 3] how are atypical PKC isozymes regulated in cells? Thus, in the next funding period, we aim to understand the cellular mechanisms of signal propagation, signal termination, and signal cross-talk using novel fluorescence technologies to 'spy'on cell signaling.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
5R37GM043154-24
Application #
8225203
Study Section
Special Emphasis Panel (NSS)
Program Officer
Chin, Jean
Project Start
1989-12-01
Project End
2013-08-31
Budget Start
2012-03-01
Budget End
2013-08-31
Support Year
24
Fiscal Year
2012
Total Cost
$410,453
Indirect Cost
$140,369
Name
University of California San Diego
Department
Pharmacology
Type
Schools of Medicine
DUNS #
804355790
City
La Jolla
State
CA
Country
United States
Zip Code
92093
Cone, Angela C; Cavin, Gabriel; Ambrosi, Cinzia et al. (2014) Protein kinase C?-mediated phosphorylation of Connexin43 gap junction channels causes movement within gap junctions followed by vesicle internalization and protein degradation. J Biol Chem 289:8781-98
Antal, Corina E; Violin, Jonathan D; Kunkel, Maya T et al. (2014) Intramolecular conformational changes optimize protein kinase C signaling. Chem Biol 21:459-69
Wu-Zhang, Alyssa X; Newton, Alexandra C (2013) Protein kinase C pharmacology: refining the toolbox. Biochem J 452:195-209
Scott, Angela M; Antal, Corina E; Newton, Alexandra C (2013) Electrostatic and hydrophobic interactions differentially tune membrane binding kinetics of the C2 domain of protein kinase C*. J Biol Chem 288:16905-15
O'Neill, Audrey K; Niederst, Matthew J; Newton, Alexandra C (2013) Suppression of survival signalling pathways by the phosphatase PHLPP. FEBS J 280:572-83
O'Neill, Audrey K; Gallegos, Lisa L; Justilien, Verline et al. (2011) Protein kinase C* promotes cell migration through a PDZ-dependent interaction with its novel substrate discs large homolog 1 (DLG1). J Biol Chem 286:43559-68
Gallegos, Lisa L; Newton, Alexandra C (2011) Genetically encoded fluorescent reporters to visualize protein kinase C activation in live cells. Methods Mol Biol 756:295-310
Warfel, Noel A; Niederst, Matt; Newton, Alexandra C (2011) Disruption of the interface between the pleckstrin homology (PH) and kinase domains of Akt protein is sufficient for hydrophobic motif site phosphorylation in the absence of mTORC2. J Biol Chem 286:39122-9
Newton, Alexandra C (2010) Protein kinase C: poised to signal. Am J Physiol Endocrinol Metab 298:E395-402
Kunkel, Maya T; Newton, Alexandra C (2010) Calcium transduces plasma membrane receptor signals to produce diacylglycerol at Golgi membranes. J Biol Chem 285:22748-52

Showing the most recent 10 out of 64 publications