Identification of three NFAT binding motifs in the 5′-upstream region of the human CD3γ gene that differentially bind NFATc1, NFATc2, and NF-κB p50

Bassam M. Badran, Steven M. Wolinsky, Arsène Burny, Karen E. Willard-Gallo*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

30 Scopus citations

Abstract

Human immunodeficiency virus, type 1 (HIV-1) infection of CD4+ T cells progressively abrogates T cell receptor (TCR)·CD3 function and surface expression by specifically interfering with CD3γ gene transcription. Our data show that the loss of CD3γ transcripts begins very early after infection and accumulates to a >90% deficiency before a significant effect on surface receptor density is apparent. Blocking TCR·CD3-directed NFAT activation with cyclosporin A provokes a partial re-expression of CD3γ gene transcripts and surface complexes in a time- and dose-dependent manner. We have identified three NFAT consensus sequences (5′-GGAAA-3′) in the 5′-upstream region of the human CD3γ gene at: -124 to -120 (NFATγ1), -384 to -380 (NFATγ2), and +450 to +454 (NFATγ3) from the first transcription initiation site. Using electrophoretic mobility shift and supershift assays, we show that NFATc2 alone binds to the NFATγ2 motif; however, complexes containing either NFATc2 or NFATc1 plus NF-κB p50 bind to the NFATγ1 and NFATγ3 sites. We further demonstrate that NFATc1 and NF-κB p50 bind in the same protein·DNA complex and that a fourth Ala added to the core sequence (5′-GGAAAA-3′) in NFATγ1, and NFATγ3 is critical for their binding. Finally, we have shown that an increase in the binding of nuclear NFATc2, NFATc1, and NF-κB p50 to these three motifs is correlated with a progressive loss of CD3γ transcripts after HIV-1 infection.

Original languageEnglish (US)
Pages (from-to)47136-47148
Number of pages13
JournalJournal of Biological Chemistry
Volume277
Issue number49
DOIs
StatePublished - Dec 6 2002

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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