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PMID: 10330401 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

PKCepsilon, via its regulatory domain and independently of its catalytic domain, induces neurite-like processes in neuroblastoma cells.

The Journal of cell biology ·Vol. 145 ·No. 4 ·1999-05-17 ·Pages 713-26

Zeidman R, Löfgren B, Pâhlman S, Larsson C

Abstract

To investigate the role of protein kinase C (PKC) isoforms in regulation of neurite outgrowth, PKCalpha, betaII, delta, and epsilon fused to enhanced green fluorescent protein (EGFP) were transiently overexpressed in neuroblastoma cells. Overexpression of PKCepsilon-EGFP induced cell processes whereas the other isoforms did not. The effect of PKCepsilon-EGFP was not suppressed by the PKC inhibitor GF109203X. Instead, process formation was more pronounced when the regulatory domain was introduced. Overexpression of various fragments from PKCepsilon regulatory domain revealed that a region encompassing the pseudosubstrate, the two C1 domains, and parts of the V3 region were necessary and sufficient for induction of processes. By deleting the second C1 domain from this construct, a dominant-negative protein was generated which suppressed processes induced by full-length PKCepsilon and neurites induced during retinoic acid- and growth factor-induced differentiation. As with neurites in differentiated neuroblastoma cells, processes induced by the PKCepsilon- PSC1V3 protein contained alpha-tubulin, neurofilament-160, and F-actin, but the PKCepsilon-PSC1V3-induced processes lacked the synaptic markers synaptophysin and neuropeptide Y. These data suggest that PKCepsilon, through its regulatory domain, can induce immature neurite-like processes via a mechanism that appears to be of importance for neurite outgrowth during neuronal differentiation.

MeSH Terms
Actins/metabolism Animals Binding Sites COS Cells Catalytic Domain Cell Division Green Fluorescent Proteins Humans Intracellular Fluid Isoenzymes/genetics,metabolism Luminescent Proteins/metabolism Neurites/physiology Neuroblastoma Protein Kinase C/genetics,metabolism Protein Kinase C-epsilon Recombinant Fusion Proteins/genetics,metabolism Tumor Cells, Cultured
Chemicals
Actins Isoenzymes Luminescent Proteins Recombinant Fusion Proteins Green Fluorescent Proteins PRKCE protein, human Protein Kinase C Protein Kinase C-epsilon
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Zeidman R
Lund University, Department of Laboratory Medicine, Molecular Medicine, Malmö University Hospital, S-205 02 Malmö, Sweden.
Löfgren B
Pâhlman S
Larsson C
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.145.4.713
Published
1999-05-17
Pages
713-26
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2133186
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.145.4.713 PMC Full Text View journal details
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