The class II phosphatidylinositol 3 kinase C2β is required for the activation of the K+ channel KCa3.1 and CD4 T-cells

Shekhar Srivastava, Lie Di, Olga Zhdanova, Zhai Li, Santosha Vardhana, Qi Wan, Ying Yan, Rajat Varma, Jonathan M. Backer, Heike Wulff, Michael L. Dustin, Edward Y. Skolnik

Research output: Contribution to journalArticle

42 Citations (Scopus)

Abstract

The Ca2+-activated K+ channel KCa3.1 is required for Ca2+ influx and the subsequent activation of T-cells. We previously showed that nucleoside diphosphate kinase beta (NDPK-B), a mammalian histidine kinase, directly phosphorylates and activates KCa3.1 and is required for the activation of human CD4 T lymphocytes. We now show that the class II phosphatidylinositol 3 kinase C2β (PI3K-C2β) is activated by the T-cell receptor (TCR) and functions upstream of NDPK-B to activate KCa3.1 channel activity. Decreased expression of PI3K-C2β by siRNA in human CD4 T-cells resulted in inhibition of KCa3.1 channel activity. The inhibition was due to decreased phosphatidylinositol 3-phosphate [PI(3)P] because dialyzing PI3K-C2β siRNA-treated T-cells with PI(3)P rescued KCa3.1 channel activity. Moreover, overexpression of PI3K-C2β in KCa3.1-transfected Jurkat T-cells led to increased TCR-stimulated activation of KCa3.1 and Ca2+ influx, whereas silencing of PI3K-C2β inhibited both responses. Using total internal reflection fluorescence microscopy and planar lipid bilayers, we found that PI3K-C2β colocalized with Zap70 and the TCR in peripheral microclusters in the immunological synapse. This is the first demonstration that a class II PI3K plays a critical role in T-cell activation.

Original languageEnglish (US)
Pages (from-to)3783-3791
Number of pages9
JournalMolecular Biology of the Cell
Volume20
Issue number17
DOIs
StatePublished - Sep 1 2009

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Class II Phosphatidylinositol 3-Kinases
Phosphatidylinositol 3-Kinase
T-Lymphocytes
T-Cell Antigen Receptor
Nucleoside-Diphosphate Kinase
Small Interfering RNA
Immunological Synapses
Calcium-Activated Potassium Channels
Jurkat Cells
Lipid Bilayers
Phosphatidylinositol 3-Kinases
Fluorescence Microscopy

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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The class II phosphatidylinositol 3 kinase C2β is required for the activation of the K+ channel KCa3.1 and CD4 T-cells. / Srivastava, Shekhar; Di, Lie; Zhdanova, Olga; Li, Zhai; Vardhana, Santosha; Wan, Qi; Yan, Ying; Varma, Rajat; Backer, Jonathan M.; Wulff, Heike; Dustin, Michael L.; Skolnik, Edward Y.

In: Molecular Biology of the Cell, Vol. 20, No. 17, 01.09.2009, p. 3783-3791.

Research output: Contribution to journalArticle

Srivastava, S, Di, L, Zhdanova, O, Li, Z, Vardhana, S, Wan, Q, Yan, Y, Varma, R, Backer, JM, Wulff, H, Dustin, ML & Skolnik, EY 2009, 'The class II phosphatidylinositol 3 kinase C2β is required for the activation of the K+ channel KCa3.1 and CD4 T-cells', Molecular Biology of the Cell, vol. 20, no. 17, pp. 3783-3791. https://doi.org/10.1091/mbc.E09-05-0390
Srivastava, Shekhar ; Di, Lie ; Zhdanova, Olga ; Li, Zhai ; Vardhana, Santosha ; Wan, Qi ; Yan, Ying ; Varma, Rajat ; Backer, Jonathan M. ; Wulff, Heike ; Dustin, Michael L. ; Skolnik, Edward Y. / The class II phosphatidylinositol 3 kinase C2β is required for the activation of the K+ channel KCa3.1 and CD4 T-cells. In: Molecular Biology of the Cell. 2009 ; Vol. 20, No. 17. pp. 3783-3791.
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