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(Received for publication, April 22, 1996, and in revised form, June 6, 1996)
From the Section of Experimental Pathology, Department of
Pathology, Roger Williams Medical Center-Brown University,
Providence, Rhode Island 02908
The cyclosporin A (CsA)/FK506-sensitive nuclear
factor of activated T cells (NFAT) plays a key role in the inducible
expression of cytokine genes in T cells. Although NFAT has been
recently shown to be inducible in several non-T immune cells, the NFAT
gene family members characterized to date have been isolated only from
T cells. To further characterize NFAT function in human B cells and to
demonstrate cytokine gene specificity of NFAT proteins, we report here
the isolation and characterization of a cDNA clone from the Raji B
cell line. The cDNA clone encodes a new isoform, NFATc.
Volume 271, Number 34,
Issue of August 23, 1996
pp. 20914-20921
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.
, of the
NFAT gene family member NFATc (designated here NFATc.
). The amino
acid sequence of NFATc. differs from that of NFATc. in the first
NH2-terminal 29 residues and contains an additional region
of 142 residues at the COOH terminus. Northern analysis using a probe
encompassing a common region of both isoforms showed two mRNA
species of 2.7 and 4.5 kilobase pairs, while an NFATc.-specific
probe detected only the 4.5-kilobase pair mRNA which was
preferentially expressed in the spleen. Transient expression of
NFATc. was capable of activating an interleukin-2 NFAT-driven
reporter gene in stimulated Jurkat cells in a CsA-sensitive manner.
However, NFATc. neither bound to the 
3 element (an NFAT-binding
site) in the tumor necrosis factor- promoter nor activated the tumor
necrosis factor- promoter in cotransfection assays. These data
suggest that different members or isoforms of NFAT gene family may
regulate inducible expression of different cytokine genes.
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