STRUCTURE, FUNCTION AND REGULATION OF PROTEIN KINASE C

Summary

Principal Investigator: Alexandra C Newton
Abstract: DESCRIPTION (provided by applicant): The long-term goal of this proposal is to understand the molecular and cellular mechanisms of protein kinase C and how these mechanisms are altered by cancer-associated mutations in members of this enzyme family to promote oncogenesis. Protein kinase C isozymes transduce the myriad of signals resulting from receptor-mediated hydrolysis of phospholipids, playing critical roles in diverse cellular functions The discovery in the 1980s that they are the receptors for the potent tumor promoting phorbol esters, led to the dogma that activation of protein kinase C by phorbol esters promotes tumorigenesis. However, the finding that long-term treatment with phorbol esters causes the degradation of protein kinase C opens this interpretation to question. Protein kinase C levels are aberrant in diverse cancers, and over 160 mutations in protein kinase C isozymes have been found in human cancers. However, whether protein kinase C isozymes function as tumor suppressors or oncogenes is not established. By understanding the molecular and cellular mechanisms of protein kinase C, and taking advantage of novel tools our lab has developed, we are poised to address this fundamental question. Thus, a central hypothesis driving this proposal is that protein kinase C may be a tumor suppressor, with inactivation (rather than activation, as is the dogma) of the kinase promoting oncogenic pathways. Three Aims are proposed: 1] to advance knowledge on the molecular mechanisms of protein kinase C, 2] to understand protein kinase C signaling in cells, and 3] to examine how cancer-associated mutations in protein kinase C alter its function, testing the hypothesis that protein kinase C may be a tumor suppressor rather than oncogene.
Funding Period: 1989-12-01 - 2017-05-31
more information: NIH RePORT

Top Publications

  1. pmc Increased membrane affinity of the C1 domain of protein kinase Cdelta compensates for the lack of involvement of its C2 domain in membrane recruitment
    Jennifer R Giorgione
    Department of Pharmacology, University of California at San Diego, La Jolla, California 92093 0721, USA
    J Biol Chem 281:1660-9. 2006
  2. pmc Genetically encoded fluorescent reporters to visualize protein kinase C activation in live cells
    Lisa L Gallegos
    Department of Pharmacology, University of California San Diego, La Jolla, CA, USA
    Methods Mol Biol 756:295-310. 2011
  3. pmc 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
    Noel A Warfel
    Department of Pharmacology, University of California at San Diego, La Jolla, California 92093, USA
    J Biol Chem 286:39122-9. 2011
  4. pmc Protein kinase Cα promotes cell migration through a PDZ-dependent interaction with its novel substrate discs large homolog 1 (DLG1)
    Audrey K O'Neill
    Department of Pharmacology, University of California, San Diego, La Jolla, California 92093, USA
    J Biol Chem 286:43559-68. 2011
  5. pmc Suppression of survival signalling pathways by the phosphatase PHLPP
    Audrey K O'Neill
    Biomedical Sciences Graduate Program, University of California, San Diego, CA 92093 0721, USA
    FEBS J 280:572-83. 2013
  6. pmc Euphohelioscopin A is a PKC activator capable of inducing macrophage differentiation
    Lorenzo de Lichtervelde
    Department of Chemistry, The Scripps Research Institute, 10550 N Torrey Pines Road, La Jolla, CA 92037, USA
    Chem Biol 19:994-1000. 2012
  7. pmc Isozyme-specific interaction of protein kinase Cδ with mitochondria dissected using live cell fluorescence imaging
    Alyssa X Wu-Zhang
    Department of Pharmacology, University of California San Diego, La Jolla, California 92093, USA
    J Biol Chem 287:37891-906. 2012
  8. pmc Electrostatic and hydrophobic interactions differentially tune membrane binding kinetics of the C2 domain of protein kinase Cα
    Angela M Scott
    Department of Pharmacology, University of California San Diego, La Jolla, California 92093 0721, USA
    J Biol Chem 288:16905-15. 2013
  9. pmc Protein kinase C pharmacology: refining the toolbox
    Alyssa X Wu-Zhang
    Department of Pharmacology, University of California, San Diego, La Jolla, CA 92093 0721, USA
    Biochem J 452:195-209. 2013
  10. pmc Spatiotemporal dynamics of phosphorylation in lipid second messenger signaling
    Corina E Antal
    Department of Pharmacology, University of California at San Diego, La Jolla, California 92093 0721
    Mol Cell Proteomics 12:3498-508. 2013

Research Grants

  1. UNMC EPPLEY CANCER CENTER SUPPORT GRANT
    Kenneth H Cowan; Fiscal Year: 2013
  2. SPORE in GI Cancer
    Robert J Coffey; Fiscal Year: 2013
  3. University of Maryland Greenebaum Cancer Center Support Grant
    Kevin J Cullen; Fiscal Year: 2013
  4. PKC/PKA Regulation in Prostate Cancer by SSeCKS/Gravin/AKAP12
    Irwin H Gelman; Fiscal Year: 2013
  5. Processing of Complex Lesions in the Mammalian Genome
    Randy J Legerski; Fiscal Year: 2013
  6. Dissecting stage-specific roles of TGF-beta in epidermal tumor progression
    Naoki Oshimori; Fiscal Year: 2013
  7. The MIT Center for Single-Cell Dynamics in Cancer (SCDC)
    Scott R Manalis; Fiscal Year: 2013
  8. Cellular Decisions of Differentiation in the GI Tract
    Juanita L Merchant; Fiscal Year: 2013
  9. The ARF Tumor Suppressor
    Maureen E Murphy; Fiscal Year: 2013
  10. The Role of PHLPP in Colon Cancer
    Tianyan Gao; Fiscal Year: 2013

Detail Information

Publications25

  1. pmc Increased membrane affinity of the C1 domain of protein kinase Cdelta compensates for the lack of involvement of its C2 domain in membrane recruitment
    Jennifer R Giorgione
    Department of Pharmacology, University of California at San Diego, La Jolla, California 92093 0721, USA
    J Biol Chem 281:1660-9. 2006
    ....
  2. pmc Genetically encoded fluorescent reporters to visualize protein kinase C activation in live cells
    Lisa L Gallegos
    Department of Pharmacology, University of California San Diego, La Jolla, CA, USA
    Methods Mol Biol 756:295-310. 2011
    ..J Biol Chem 281:30947-30956, 2006). Here we describe methods to image cells expressing the reporters and elaborate on data analyses, control experiments, and variations for imaging the activity of expressed PKC...
  3. pmc 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
    Noel A Warfel
    Department of Pharmacology, University of California at San Diego, La Jolla, California 92093, USA
    J Biol Chem 286:39122-9. 2011
    ..In summary, these data support a model in which Akt can be phosphorylated at Ser-473 and activated in the absence of mTORC2 by mechanisms that depend on removal of the PH domain from the kinase domain...
  4. pmc Protein kinase Cα promotes cell migration through a PDZ-dependent interaction with its novel substrate discs large homolog 1 (DLG1)
    Audrey K O'Neill
    Department of Pharmacology, University of California, San Diego, La Jolla, California 92093, USA
    J Biol Chem 286:43559-68. 2011
    ..Taken together, these data establish the requirement of scaffolding to DLG1 for PKCα to promote cellular migration...
  5. pmc Suppression of survival signalling pathways by the phosphatase PHLPP
    Audrey K O'Neill
    Biomedical Sciences Graduate Program, University of California, San Diego, CA 92093 0721, USA
    FEBS J 280:572-83. 2013
    ..This review summarizes the targets and cellular actions of PHLPP, with emphasis on its role as a tumour suppressor in the oncogenic phosphoinositide 3-kinase (PI3K)/Akt signalling cascade...
  6. pmc Euphohelioscopin A is a PKC activator capable of inducing macrophage differentiation
    Lorenzo de Lichtervelde
    Department of Chemistry, The Scripps Research Institute, 10550 N Torrey Pines Road, La Jolla, CA 92037, USA
    Chem Biol 19:994-1000. 2012
    ..In addition to shedding insights into normal hematopoiesis, this work may ultimately facilitate the application of stem cell therapies to a host of myeloid dysfunctions...
  7. pmc Isozyme-specific interaction of protein kinase Cδ with mitochondria dissected using live cell fluorescence imaging
    Alyssa X Wu-Zhang
    Department of Pharmacology, University of California San Diego, La Jolla, California 92093, USA
    J Biol Chem 287:37891-906. 2012
    ..These data support a model in which multiple determinants unique to PKCδ drive a specific interaction with mitochondria that promotes mitochondrial respiration...
  8. pmc Electrostatic and hydrophobic interactions differentially tune membrane binding kinetics of the C2 domain of protein kinase Cα
    Angela M Scott
    Department of Pharmacology, University of California San Diego, La Jolla, California 92093 0721, USA
    J Biol Chem 288:16905-15. 2013
    ....
  9. pmc Protein kinase C pharmacology: refining the toolbox
    Alyssa X Wu-Zhang
    Department of Pharmacology, University of California, San Diego, La Jolla, CA 92093 0721, USA
    Biochem J 452:195-209. 2013
    ....
  10. pmc Spatiotemporal dynamics of phosphorylation in lipid second messenger signaling
    Corina E Antal
    Department of Pharmacology, University of California at San Diego, La Jolla, California 92093 0721
    Mol Cell Proteomics 12:3498-508. 2013
    ....
  11. pmc Imaging kinase activity at protein scaffolds
    Maya T Kunkel
    Department of Pharmacology, University of California at San Diego, San Diego, CA, USA
    Methods Mol Biol 1071:129-37. 2014
    ..This protocol describes how to assay kinase activity at a protein scaffold in live cells using a fluorescence resonance energy transfer (FRET)-based kinase activity sensor for protein kinase D (PKD) as an example. ..
  12. pmc Identification of PHLPP1 as a tumor suppressor reveals the role of feedback activation in PTEN-mutant prostate cancer progression
    Muhan Chen
    Cold Spring Harbor Laboratory, Cold Spring Harbor, NY 11724, USA
    Cancer Cell 20:173-86. 2011
    ..These data establish a conceptual framework for progression of PTEN mutant prostate cancer to life-threatening disease...
  13. pmc Active site inhibitors protect protein kinase C from dephosphorylation and stabilize its mature form
    Christine M Gould
    Department of Pharmacology, University of California, San Diego, La Jolla, California 92093 0721, USA
    J Biol Chem 286:28922-30. 2011
    ..These data provide a molecular explanation for the recent studies showing that active site inhibitors stabilize the phosphorylation state of protein kinases B/Akt and C...
  14. pmc Protein kinase C {delta}-specific activity reporter reveals agonist-evoked nuclear activity controlled by Src family of kinases
    Taketoshi Kajimoto
    Department of Pharmacology, University of California at San Diego, La Jolla, California 92093, USA
    J Biol Chem 285:41896-910. 2010
    ....
  15. ncbi Targeting protein kinase C activity reporter to discrete intracellular regions reveals spatiotemporal differences in agonist-dependent signaling
    Lisa L Gallegos
    Department of Pharmacology, University of California at San Diego, La Jolla, California 92093, USA
    J Biol Chem 281:30947-56. 2006
    ..Our data reveal that the magnitude and duration of PKC signaling is location-specific and controlled by the level of phosphatase activity and persistence of DAG at each location...
  16. ncbi A single residue in the C1 domain sensitizes novel protein kinase C isoforms to cellular diacylglycerol production
    Daniel R Dries
    Biomedical Sciences Graduate Program and Department of Pharmacology, University of California at San Diego, La Jolla, California 92093, USA
    J Biol Chem 282:826-30. 2007
    ..The finding that a single residue controls the affinity of the C1 domain for DAG-containing membranes provides a molecular explanation for why 1) DAG alone is sufficient to activate nPKCs but not cPKCs and 2) nPKCs target to the Golgi...
  17. ncbi The phosphatase PHLPP controls the cellular levels of protein kinase C
    Tianyan Gao
    Department of Pharmacology, University of California at San Diego, La Jolla, CA 92093, USA
    J Biol Chem 283:6300-11. 2008
    ..These data reveal that PHLPP controls the cellular levels of PKC by specifically dephosphorylating the hydrophobic motif, thus destabilizing the enzyme and promoting its degradation...
  18. ncbi Kinetic analysis of the interaction of the C1 domain of protein kinase C with lipid membranes by stopped-flow spectroscopy
    Daniel R Dries
    Biomedical Sciences Graduate Program, University of California at San Diego, La Jolla, CA 92093, USA
    J Biol Chem 283:7885-93. 2008
    ....
  19. pmc PHLiPPing the switch on Akt and protein kinase C signaling
    John Brognard
    Department of Pharmacology, University of California, San Diego, La Jolla, CA 92093 0721, USA
    Trends Endocrinol Metab 19:223-30. 2008
    ..This review focuses on the function of PHLPP1 and PHLPP2 in modulating signaling by Akt and PKC...
  20. pmc Lipid activation of protein kinases
    Alexandra C Newton
    Department of Pharmacology, University of California at San Diego, La Jolla, CA 92093 0721, USA
    J Lipid Res 50:S266-71. 2009
    ....
  21. pmc The chaperones Hsp90 and Cdc37 mediate the maturation and stabilization of protein kinase C through a conserved PXXP motif in the C-terminal tail
    Christine M Gould
    Pharmacology Department, University of California, San Diego, La Jolla, California 92039 0721, USA
    J Biol Chem 284:4921-35. 2009
    ....
  22. pmc Protein kinase C: poised to signal
    Alexandra C Newton
    Dept of Pharmacology, Univ of California at San Diego, La Jolla, 92093, USA
    Am J Physiol Endocrinol Metab 298:E395-402. 2010
    ..This review introduces the PKC family and then focuses on recent advances in understanding the cellular regulation of its diacylglycerol-regulated members...
  23. pmc Calcium transduces plasma membrane receptor signals to produce diacylglycerol at Golgi membranes
    Maya T Kunkel
    Department of Pharmacology, University of California at San Diego, La Jolla, California 92093, USA
    J Biol Chem 285:22748-52. 2010
    ..Thus, agonist-evoked increases in intracellular Ca(2+) cause increases in Golgi diacylglycerol, allowing this intracellular membrane to serve as a platform for signaling by protein kinases C and D...
  24. pmc Intramolecular conformational changes optimize protein kinase C signaling
    Corina E Antal
    Department of Pharmacology, University of California at San Diego, La Jolla, CA 92093, USA Biomedical Sciences Graduate Program, University of California at San Diego, La Jolla, CA 92093, USA
    Chem Biol 21:459-69. 2014
    ..The conformational rearrangements of PKC serve as a paradigm for how multimodule transducers optimize their dynamic range of signaling. ..

Research Grants30

  1. UNMC EPPLEY CANCER CENTER SUPPORT GRANT
    Kenneth H Cowan; Fiscal Year: 2013
    ....
  2. SPORE in GI Cancer
    Robert J Coffey; Fiscal Year: 2013
    ..Administrative Core B. Translational Pathology &Imaging Core C. Biostatistics &Bioinformatics Career Development Program Developmental Research Program ..
  3. University of Maryland Greenebaum Cancer Center Support Grant
    Kevin J Cullen; Fiscal Year: 2013
    ..Reflecting our remarkable and continued growth, UMGCC seeks to renew its CCSG to enhance and expand its efforts in high-quality and clinically relevant cancer research. ..
  4. PKC/PKA Regulation in Prostate Cancer by SSeCKS/Gravin/AKAP12
    Irwin H Gelman; Fiscal Year: 2013
    ..These studies will elucidate how the loss of SSeCKS expression in prostate cancer increases disease progression by allowing PKC to be hyperactivated and dysregulated. ..
  5. Processing of Complex Lesions in the Mammalian Genome
    Randy J Legerski; Fiscal Year: 2013
    ..These approaches have excellent potential to yield useful technical and therapeutic advances in genetic manipulation. ..
  6. Dissecting stage-specific roles of TGF-beta in epidermal tumor progression
    Naoki Oshimori; Fiscal Year: 2013
    ..The many resources in the Fuchs lab and university resource centers will provide equipment, training, and technical expertise that will ensure successful completion of the proposed research. ..
  7. The MIT Center for Single-Cell Dynamics in Cancer (SCDC)
    Scott R Manalis; Fiscal Year: 2013
    ..These facilities and all reagents generated by the cores will be made available to other PS-OCs. ..
  8. Cellular Decisions of Differentiation in the GI Tract
    Juanita L Merchant; Fiscal Year: 2013
    ....
  9. The ARF Tumor Suppressor
    Maureen E Murphy; Fiscal Year: 2013
    ..We believe that the requirement for ARF and autophagy may be an Achilles Heel for tumor cells. This research is aimed at understanding this pathway, and at finding ways to manipulate this pathway to combat cancer. ..
  10. The Role of PHLPP in Colon Cancer
    Tianyan Gao; Fiscal Year: 2013
    ....
  11. Autophagy and Epithelial Cell Fate During Anoikis and 3D Morphogenesis
    Jayanta Debnath; Fiscal Year: 2013
    ..Ultimately, these studies may lead to new treatment strategies to prevent the uncontrolled expansion of cancer cells via manipulating autophagy. ..
  12. Regulation of K-Ras by a Farnesyl-electrostatic Switch
    MARK REID PHILIPS; Fiscal Year: 2013
    ..We propose to study the cell biology and physiology of K-Ras phosphorylation to better understand how to exploit this process to develop anti-cancer drugs. ..
  13. Signaling and Metabolic Control in the Drosophila Eye
    Utpal Banerjee; Fiscal Year: 2013
    ..In AIM 3, the oncogenic influence on cellular metabolism and the molecular mechanism that causes a metabolic shift towards glycolysis will be investigated. ..
  14. The Mechanism of Nur77 and Nor1 Induced Apoptosis Through Interaction with Bcl2
    MEGAN LINN BURGER; Fiscal Year: 2013
    ....
  15. Molecular Analysis and Role of RGS6 as a Novel Growth Suppressor
    Rory A Fisher; Fiscal Year: 2013
    ..They will provide new understanding of the pathogenesis of urinary bladder cancer and potentially identify RGS6 as a biomarker for the prognosis or diagnosis of bladder cancer as well as a new therapeutic target for its treatment. ..
  16. Mining B-catenin/BCL9 transcriptional complex for Multiple Myeloma therapeutics
    Ruben D Carrasco; Fiscal Year: 2013
    ..abstract_text> ..
  17. Mechanisms of GVHD
    Joseph H Antin; Fiscal Year: 2013
    ..Ultimately we envision an integrated genomic profile that will determine the type of GVHD prophylaxis that will be most effective. ..
  18. CSHL CANCER CENTER SUPPORT GRANT
    BRUCE W STILLMAN; Fiscal Year: 2013
    ....