Inhibition of NF-AT signal transduction events by a dominant-negative form of calcineurin

Taro Muramatsu, Randall L. Kincaid

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

An inhibitory, 'dominant-negative,' form of the calcineurin catalytic (A) subunit was prepared, which lacks the calmodulin-binding domain, autoinhibitory domain and most of its catalytic core but possesses die regulatory (B) subunit binding domain. When tested for its ability to block calcineurin-dependent signaling in Jurkat cells, expression of this 'B-subunit knock-out' (BKO) construct suppressed reporter gene activity driven by NF-AT, the pivotal promoter element for interleukin (IL)-2 gene induction. Immunoprecipitation of epitope-labeled BKO demonstrated for the formation of a tight complex with endogenous B subunit in Jurkat cells, consistent with an inhibitory mechanism that involves the sequestration of the B subunit. Furthermore, the sharply reduced NF-AT activity produced by co-transfecting BKO could be 'rescued' by overexpression of transfected B subunit, suggesting that depletion of this subunit was responsible for the inhibition. These data suggest the potential utility of agents that disrupt calcineurin-mediated signal transduction pathways by blocking formation of the catalytically active dimer of calcineurin A and B subunits.

Original languageEnglish
Pages (from-to)466-472
Number of pages7
JournalBiochemical and Biophysical Research Communications
Volume218
Issue number2
DOIs
Publication statusPublished - 1996 Jan 17
Externally publishedYes

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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