TY - JOUR
T1 - Enhancer role of STAT5 in CD2 activation of IFN-γ gene expression
AU - Gonsky, Rivkah
AU - Deem, Richard L.
AU - Bream, Jay
AU - Young, Howard A.
AU - Targan, Stephan R.
PY - 2004/11/15
Y1 - 2004/11/15
N2 - IFN-γ is an important immunoregulatory protein with tightly controlled expression in activated T and NK cells. Three potential STAT binding regions have been recognized within the IFN-γ promoter: 1) an IL-12-mediated STAT4 binding site at -236 bp; 2) a newly identified IL-2-induced STAT5 binding element at -3.6 kb; and 3) CD2-mediated STAT1 and STAT4 binding to an intronic element in mucosal T cells. However, functional activation of these sites remains unclear. In this study we demonstrate CD2-mediated activation of the newly characterized -3.6-kb IFN-γ STAT5 binding region. CD2 signaling of human PBMC results in activation of the -3.6-kb IFN-γ promoter, whereas mutation of the -3.6-kb STAT5 site attenuates promoter activity. Functional activation is accompanied by STAT5A but little STAT5B nucleoprotein binding to the IFN-γ STAT5 site, as determined by competition and supershift assays. STAT5 activation via CD2 occurs independent of IL-2. Western and FACS analysis shows increased phospho-STAT5 following CD2 signaling. AG490, a tyrosine kinase inhibitor affecting Jak proteins, inhibits CD2-mediated IFN-γ mRNA expression, secretion, and nucleoprotein binding to the IFN-γ STAT5 site in a dose-dependent fashion. This report is the first to describe CD2-mediated activation of STAT5 and supports STAT5 involvement in regulation of IFN-γ expression.
AB - IFN-γ is an important immunoregulatory protein with tightly controlled expression in activated T and NK cells. Three potential STAT binding regions have been recognized within the IFN-γ promoter: 1) an IL-12-mediated STAT4 binding site at -236 bp; 2) a newly identified IL-2-induced STAT5 binding element at -3.6 kb; and 3) CD2-mediated STAT1 and STAT4 binding to an intronic element in mucosal T cells. However, functional activation of these sites remains unclear. In this study we demonstrate CD2-mediated activation of the newly characterized -3.6-kb IFN-γ STAT5 binding region. CD2 signaling of human PBMC results in activation of the -3.6-kb IFN-γ promoter, whereas mutation of the -3.6-kb STAT5 site attenuates promoter activity. Functional activation is accompanied by STAT5A but little STAT5B nucleoprotein binding to the IFN-γ STAT5 site, as determined by competition and supershift assays. STAT5 activation via CD2 occurs independent of IL-2. Western and FACS analysis shows increased phospho-STAT5 following CD2 signaling. AG490, a tyrosine kinase inhibitor affecting Jak proteins, inhibits CD2-mediated IFN-γ mRNA expression, secretion, and nucleoprotein binding to the IFN-γ STAT5 site in a dose-dependent fashion. This report is the first to describe CD2-mediated activation of STAT5 and supports STAT5 involvement in regulation of IFN-γ expression.
UR - http://www.scopus.com/inward/record.url?scp=8444243255&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=8444243255&partnerID=8YFLogxK
U2 - 10.4049/jimmunol.173.10.6241
DO - 10.4049/jimmunol.173.10.6241
M3 - Article
C2 - 15528362
AN - SCOPUS:8444243255
SN - 0022-1767
VL - 173
SP - 6241
EP - 6247
JO - Journal of Immunology
JF - Journal of Immunology
IS - 10
ER -