Document Detail


Regulation of receptor-mediated protein kinase C membrane trafficking by autophosphorylation.
MedLine Citation:
PMID:  10828076     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Signal transduction via protein kinase C (PKC) is closely regulated by its subcellular localization. In response to activation of cell-surface receptors, PKC is directed to the plasma membrane by two membrane-targeting domains, namely the C1 and C2 regions. This is followed by the return of the enzyme to the cytoplasm, a process shown recently to require PKC autophosphorylation (Feng, X., and Hannun, Y. A. (1998) J. Biol. Chem. 273, 26870-26874). In the present study, we examined mechanisms of translocation and reverse translocation and the role of autophosphorylation in these processes. By visualizing the trafficking of wild-type as well as mutant PKCbetaII in live cells, we demonstrated that in response to cell-surface receptor activation, the function of the C1 region is required but not sufficient for recruitment of the enzyme to the plasma membrane. The C2 region is also critical in anchoring the enzyme to the plasma membrane. Furthermore, the inability of a kinase-deficient PKC to undergo reverse translocation was restored by the addition of intracellular calcium chelators, suggesting a role for the C2 region in the persistent phase of translocation. On the other hand, the inability of a C2 deletion mutant (C1 region intact) to translocate in response to agonist was reversed in mutants lacking kinase activity or by mutation of the Ser(660) autophosphorylation site to alanine, suggesting that autophosphorylation of this site is required for opposing the action of the C2 region. Therefore, the membrane-targeting function of the C1 region is facilitated by the C2 region and appears to be opposed by autophosphorylation. Taken together, these findings provide novel evidence of the functional regulation of reversible PKC membrane localization by autophosphorylation, and they show that the dynamic translocation of PKC in response to agonists is tightly regulated in a collaborative fashion by the C1 and C2 regions in balance with the effects of autophosphorylation.
Authors:
X Feng; K P Becker; S D Stribling; K G Peters; Y A Hannun
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  The Journal of biological chemistry     Volume:  275     ISSN:  0021-9258     ISO Abbreviation:  J. Biol. Chem.     Publication Date:  2000 Jun 
Date Detail:
Created Date:  2000-07-11     Completed Date:  2000-07-11     Revised Date:  2007-11-15    
Medline Journal Info:
Nlm Unique ID:  2985121R     Medline TA:  J Biol Chem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  17024-34     Citation Subset:  IM    
Affiliation:
Departments of Cell Biology and Medicine, Duke University Medical Center, Durham, North Carolina 27710, USA.
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MeSH Terms
Descriptor/Qualifier:
Biological Transport
Cell Line
Cell Membrane / enzymology
Green Fluorescent Proteins
Humans
Luminescent Proteins / metabolism
Phosphorylation
Protein Kinase C / metabolism*
Receptors, Cell Surface / metabolism
Grant Support
ID/Acronym/Agency:
HL-43707/HL/NHLBI NIH HHS
Chemical
Reg. No./Substance:
0/Luminescent Proteins; 0/Receptors, Cell Surface; 147336-22-9/Green Fluorescent Proteins; EC 2.7.11.13/Protein Kinase C

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