Direct and Indirect Inhibition of PKC

doi:10.1126/stke.2003.175.tw120

Account Information

Guest Alerts | Access Rights | My Account | Sign In

Site Navigation

Article Views

Abstract
Print View

Article Tools

Help & Feedback

My Science Signaling

Sci. STKE, 25 March 2003
Vol. 2003, Issue 175, p. tw120
[DOI: 10.1126/stke.2003.175.tw120]

EDITORS' CHOICE

PROTEIN INTERACTIONS Direct and Indirect Inhibition of PKC

Protein kinase K (PKC) is activated by the lipid diacylglycerol (DAG). DAG is metabolized by DAG kinases (DGKs), which convert DAG to phophatidic acid. Thus, DGKs control PKC activity indirectly by eliminating an allosteric activator, DAG. Both DGKs and PKCs exist as large families with variable domains involved in localization and regulation of these enzymes. Luo et al. report a novel direct mechanism for inhibition of PKC ${alpha}$ by DGK ${zeta}$ , which has a region homologous to the phosphorylation site domain of the myristoylated alanine-rich C kinase substrate (MARCKS) protein. MARCKS is a substrate for PKC, and phosphorylation of the MARCKS domain of DGK ${zeta}$ by PKC reduces nuclear localization of DGK ${zeta}$ . DGK ${zeta}$ and PKC ${alpha}$ coprecipitated from transfected cells, as did endogenous proteins expressed in rat brain extracts and nontransfected cells. The interaction was mediated by a portion of the catalytic domain of DGK ${zeta}$ . Activation of PKC by treatment of cells or recombinant PKC with phorbol esters inhibited the interaction of PKC and DGK ${zeta}$ . Activation of PKC by growth factor stimulation of cells also inhibited the coprecipitation of DGK activity. Mutations that mimicked phosphorylation of the MARCKS domain of DGK ${zeta}$ prevented the interaction with PKC, and stimulation of PKC with phorbol ester did not block the interaction between PKC and a mutant of DGK ${zeta}$ with a nonphosphorylatable MARCKS domain. Thus, phosphorylation of the MARCKS domain by PKC appears to represent one mechanism controlling the interaction between these two proteins. The importance of phosphorylation of the MARCKS domain in controlling not just protein interaction, but also DGK ${zeta}$ inhibition of PKC ${alpha}$ activity was confirmed by in vitro kinase assays of PKC immunoprecipitated from cells expressing wild-type, phosphorylation-mimic versions of DGK ${zeta}$ , or nonphosphorylatable versions of DGK ${zeta}$ . The authors propose a model whereby in the basal state, the DGK ${zeta}$ -PKC ${alpha}$ complex holds PKC in an inactive state and DGK ${zeta}$ phosphorylates DAG, keeping the system off. Upon stimulation, DAG levels rise above that which DGK ${zeta}$ can metabolize, stimulating PKC ${alpha}$ activity. This results in the phosphorylation and release of DGK ${zeta}$ , further promoting PKC ${alpha}$ activity.

B. Luo, S. M. Prescott, M. K. Topham, Association of diacylglycerol kinase ${zeta}$ with protein kinase C ${alpha}$ : Spatial regulation of diacylglycerol signaling. J. Cell Biol. 160, 929-937 (2003). [Abstract] [Full Text]

Citation: Direct and Indirect Inhibition of PKC. Sci. STKE 2003, tw120 (2003).

The editors suggest the following Related Resources on Science sites:

VIRTUAL JOURNAL Association of diacylglycerol kinase ${zeta}$ with protein kinase C ${alpha}$ : spatial regulation of diacylglycerol signaling: Bai Luo, Stephen M. Prescott, and Matthew K. Topham (17 March 2003)
J. Cell Biol. 160 (6), 929. [DOI: 10.1083/jcb.200208120]
| Abstract » | Full Text » | PDF »