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Comparison of the PKCalpha and the PKCepsilon C1b Domains: Identification of Residues Critical for PKCepsilon-mediated Neurite Induction.

In English

Författare

Summary, in English

We showed earlier that over-expression of protein kinase C (PKC) epsilon induces neurite outgrowth. The effect is mediated by a region (PKC epsilon PSC1V3) encompassing the pseudosubstrate, the two C1 domains and part of the V3 region, and is independent of the catalytic activity of the enzyme. In this region, residues immediately N-terminal of the C1b domain are crucial for neurite outgrowth. However, in this study we show that the PKC epsilon C1b domain itself is necessary for neurite induction, since a mutant in which the PKC epsilon C1b domain has been replaced with the C1b domain from PKC alpha, PKC epsilon PSC1a(alpha C1b)V3 lacks neurite-inducing capacity. The molecular basis for the importance of the PKC epsilon C1b domain was investigated by mutation studies of the PKC alpha C1b domain. Point mutations were done in the PKC alpha C1b domain of the PKC epsilon PSC1a(alpha C1b)V3 construct, in which the PKC alpha residues were mutated into the corresponding residues in PKCE. This highlighted residues in the C-terminal part of the primary sequence of the C1b domain, located in the base of the Clb domain, as important for neurite outgrowth. The mutations S48P, D32K and L49N all influenced neurite induction positively. Furthermore, the mutation of L49N alone was sufficient to make PKC epsilon PSC1a(alpha C1b)V3 neuritogenic in phorbol ester-stimulated cells, and mutation of this residue in full-length PKC epsilon into the corresponding residue in PKC alpha, N291L reduced the neurite-inducing effect of PKC alpha. In conclusion, we have identified residues in the PKC epsilon C1b domain, in particular Asn49, that are essential for neurite outgrowth. (c) 2007 Elsevier Ltd. All rights reserved.

Publiceringsår

2007

Språk

Engelska

Sidor

951-965

Publikation/Tidskrift/Serie

Journal of Molecular Biology

Volym

368

Issue

4

Dokumenttyp

Artikel i tidskrift

Förlag

Elsevier

Ämne

  • Cancer and Oncology

Nyckelord

  • neuroblastoma cells
  • C1 domains
  • neurite outgrowth
  • protein kinase C

Status

Published

ISBN/ISSN/Övrigt

  • ISSN: 1089-8638