Holden, Neil S., Squires, Paul E., Kaur, Manminder, Bland, R. (Rosemary), Jones, Carol E. and Newton, R. (Robert) (2008) Phorbol ester-stimulated NF-kappa B-dependent transcription : Roles for isoforms of novel protein kinase C. Cellular Signalling, Volume 20 (Number 7). pp. 1338-1348. doi:10.1016/j.cellsig.2008.03.001

Research output not available from this repository, contact author.

Request Changes to record.

Abstract

Since protein kinase C (PKC) isoforms are variously implicated in the activation of NF-kappa B, we have investigated the role of PKC in the activation of NF-kappa B-dependent transcription by the diacyl glycerol (DAG) mimetic, phorbol 12-myristate 13-acetate (PMA), and by tumour necrosis factor (TNF) a in pulmonary A549 cells. The PKC selective inhibitors, Ro31-8220, Go6976, GF109203X and Go6983, revealed no effect on TNF alpha-induced NF kappa B DNA binding and a similar lack of effect on serine 32/36 phosphorylated I kappa B alpha and the loss of total I kappa B alpha, indicates that activation of the core IKK-I kappa B alpha-NF-kappa B cascade by TNF alpha does not involve PKC. In contrast, differential sensitivity of an NF-kappa B-dependent reporter to Ro31-8220, Go6976, GF109203X and Go6983 (EC50S 0.46 mu M, 0.34 mu M, > 10 mu M and > 10 mu M respectively) suggests a role for protein kinase D in transcriptional activation by TNF alpha. Compared with TNFa, PMA weakly induces NF-kappa B DNA binding and this effect was not associated with serine 32/36 phosphorylation of I kappa B alpha. However, PMA-stimulated NF-kappa B DNA binding was inhibited by Ro31-8220 (10 mu M), GF109203X (10 mu M) and Go6983 (10 mu M), but not by Go6976 (10 mu M), suggesting a role for novel PKCisoforms. Furthermore,a lack of positive effect of calcium mobilising agents on both NF-kappa B DNA binding and on transcriptional activation argues against major roles for classical PKCs. This, combined with the ability of both GF109203X and Go6983 to prevent enhancement of TNF alpha-induced NF-kappa B-dependent transcription by PMA, further indicates a role for novel PKCs in NF-kappa B transactivation. Finally, siRNA-mediated knockdown of PKC delta and epsilon, expression did not affect TNFa-induced NF-kappa B-dependent transcription. However, knockdown of PKC delta expression significantly inhibited PMA-stimulated luciferase activity, whereas knockdown of PKC epsilon was without effect. Furthermore, combined knockdown of PKC delta and E revealed an increased inhibitory effect on PMA-stimulated NF-kappa B-dependent transcription suggesting that PMA-induced NF-kappa B-dependent transcription is driven by novel PKC isoforms, particularly PKC delta and epsilon. (c) 2008 Elsevier Inc. All rights reserved.

Item Type:	Journal Article
Subjects:	Q Science > QH Natural history > QH426 Genetics Q Science > QP Physiology
Divisions:	Faculty of Science > Life Sciences (2010- ) > Biological Sciences ( -2010)
Library of Congress Subject Headings (LCSH):	NF-kappa B (DNA-binding protein), Protein kinase C, Transcription factors, Pulmonary endothelium, Epithelium, Gene expression, Phorbol esters
Journal or Publication Title:	Cellular Signalling
Publisher:	Elsevier
ISSN:	0898-6568
Official Date:	July 2008
Dates:	Date Event July 2008 Published
Volume:	Volume 20
Number:	Number 7
Number of Pages:	11
Page Range:	pp. 1338-1348
DOI:	10.1016/j.cellsig.2008.03.001
Status:	Peer Reviewed
Publication Status:	Published
Access rights to Published version:	Restricted or Subscription Access
Funder:	Novartis Pharmaceuticals UK, Canadian Institutes of Health Research (CIHR), Alberta Heritage Foundation for Medical Research (AHFMR)

Data sourced from Thomson Reuters' Web of Knowledge

Request changes or add full text files to a record

Repository staff actions (login required)

View Item