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PKCδ localization at the membrane causes increased cytoskeleton reorganization associated with cell spreading and protrusions
Author(s) -
Jamison Joshua A,
Wells Alan
Publication year - 2011
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.25.1_supplement.930.7
Subject(s) - protein kinase c , microbiology and biotechnology , cytoskeleton , phosphorylation , motility , actin cytoskeleton , transfection , chemistry , biology , cell , cell culture , biochemistry , genetics
In wound repair, fibroblasts coordinate asymmetric signaling for cell motility and contractions of the extracellular matrix. Systems analyses show a central role in motility for transcellular contractility as a process involving PKCδ as a mediator. In addition, PIP2 hydrolysis by PLCγ1 activates novel protein kinase c delta (PKCδ), which leads to phosphorylation of MLC‐2. As PLCγ1 is immediately activated by chemo‐kinetic signals from EGFR at the membrane, the partial activation of PKCδ would be closely localized to its activity. To directly investigate PKCδ activation and its association to MLC‐2, PKCδ constructs were created with farnesylation moieties targeting PKCδ to the membrane, denoted as (PKCδ‐CaaX) and a non‐targeting control (PKCδ‐SaaX). NR6‐WT fibroblast with PKCδ‐CaaX displayed a protrusive and expansive cell phenotype, whereas cells with PKCδ‐SaaX were similar in morphology to cells expressing GFP (transfection control). These results were further supported by PKCδ‐CaaX increased redistrubition to the leading edge with activation of phospho‐MLC and actin redistribution. From these results, it appears that membrane tethering of PKCδ leads to direct activation of MLC‐2 to affect cell morphology; interestingly, this does not require global MLC2 phosphorylation. As PKCδ localizes to the membrane, the mechanism driving cytoskeleton reorganization is dependent on membrane activated PKCδ.