Protein kinase Cα (PKCα) acts upstream of PKCθ to activate IκB kinase and NF-κB in T lymphocytes

Sergey A. Trushin, Kevin N. Pennington, Eva M Carmona Porquera, Susana Asin, Doris N. Savoy, Daniel D Billadeau, Carlos V. Paya

Research output: Contribution to journalArticle

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Abstract

NF-κB is an ubiquitous transcription factor that is a key in the regulation of the immune response and inflammation. T-cell receptor (TCR) cross-linking leads to NF-κB activation, an IκB kinase (IKK)-dependent process. However, the upstream kinases that regulate IKK activity following TCR activation remain to be fully characterized. Herein, we demonstrate using genetic analysis, pharmacological inhibition, and RNA interference (RNAi) that the conventional protein kinase C (PKC) isoform PKCα, but not PKCβ1, is required for the activation of the IKK complex following T-cell activation triggered by CD3/CD28 cross-linking. We find that in the presence of Ca2+ influx, the catalytically active PKCαA25E induces IKK activity and NF-κB-dependent transcription; which is abrogated following the mutations of two aspartates at positions 246 and 248, which are required for Ca2+ binding to PKCα and cell membrane recruitment. Kinetic studies reveal that an early phase (1 to 5 min) of IKK activation following TCR/CD28 cross-linking is PKCα dependent and that a later phase (5 to 25 min) of IKK activation is PKCθ dependent. Activation of IKK- and NF-κB-dependent transcription by PKCαA25E is abrogated by the PKCθ inhibitor rottlerin or the expression of the kinase-inactive form of PKCθ. Taken together, our results suggest that PKCα acts upstream of PKCθ to activate the IKK complex and NF-κB in T lymphocytes following TCR activation.

Original languageEnglish (US)
Pages (from-to)7068-7081
Number of pages14
JournalMolecular and Cellular Biology
Volume23
Issue number19
DOIs
StatePublished - Oct 2003

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Protein Kinase C
Phosphotransferases
T-Lymphocytes
T-Cell Antigen Receptor
Protein C Inhibitor
Protein Kinase Inhibitors
RNA Interference
Aspartic Acid
Carrier Proteins
Protein Isoforms
Transcription Factors
Cell Membrane
Pharmacology
Inflammation
Mutation

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

Protein kinase Cα (PKCα) acts upstream of PKCθ to activate IκB kinase and NF-κB in T lymphocytes. / Trushin, Sergey A.; Pennington, Kevin N.; Carmona Porquera, Eva M; Asin, Susana; Savoy, Doris N.; Billadeau, Daniel D; Paya, Carlos V.

In: Molecular and Cellular Biology, Vol. 23, No. 19, 10.2003, p. 7068-7081.

Research output: Contribution to journalArticle

Trushin, Sergey A. ; Pennington, Kevin N. ; Carmona Porquera, Eva M ; Asin, Susana ; Savoy, Doris N. ; Billadeau, Daniel D ; Paya, Carlos V. / Protein kinase Cα (PKCα) acts upstream of PKCθ to activate IκB kinase and NF-κB in T lymphocytes. In: Molecular and Cellular Biology. 2003 ; Vol. 23, No. 19. pp. 7068-7081.
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