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Nucleic Acids Research Advance Access published online on March 27, 2007
Nucleic Acids Research, doi:10.1093/nar/gkm117
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© 2007 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Molecular biology

Dynamic binding of Ku80, Ku70 and NF90 to the IL-2 promoter in vivo in activated T-cells

Lingfang Shi1, Daoming Qiu1, Guohua Zhao1, Blaise Corthesy2, Susan Lees-Miller3, Westley H. Reeves4 and Peter N. Kao1,*

1Pulmonary and Critical Care Medicine, Stanford University Medical Center, Stanford, CA 94305-5236, USA, 2Immunology and Allergy, Internal Medicine Department, University of Lausanne, CH-1011, Lausanne, Switzerland, 3Department of Biochemistry & Molecular Biology, University of Calgary, 3330 Hospital Drive NW, Calgary, Alta., Canada T2N 4N1 and 4Division of Rheumatology and Clinical Immunology, University of Florida, Gainesville, FL 32610-0211, USA

*To whom correspondence should be addressed. Tel: +650-725-0570; Fax: +650-725-5489; Email: peterkao{at}stanford.edu

Received November 20, 2006. Revised February 7, 2007. Accepted February 8, 2007.

IL-2 gene expression in activated T-cells is initiated by chromatin remodeling at the IL-2 proximal promoter and conversion of a transcriptional repressor into a potent transcriptional activator. A purine-box regulator complex was purified from activated Jurkat T-cell nuclei based on sequence-specific DNA binding to the antigen receptor response element (ARRE)/nuclear factor of activated T-cells (NF-AT) target DNA sequence in the proximal IL-2 promoter. ARRE DNA-binding subunits were identified as NF90, NF45 and systemic lupus erythematosis autoantigens, Ku80 and Ku70. Monoclonal antibodies to Ku80, Ku70 and NF90 specifically inhibit constitutive and inducible ARRE DNA-binding activity in Jurkat T-cells. Ku80, Ku70 and NF90 bind specifically to the IL-2 gene promoter in vivo, as demonstrated by chromatin immunoprecipitation. Activation of Jurkat T-cells and mouse primary spleen cells induces binding of Ku80 and NF90 to the IL-2 promoter in vivo, and decreases binding of Ku70 to the IL-2 promoter in vivo, and these dynamic changes are inhibited by immunosuppressants cyclosporin A and triptolide. Dynamic changes in binding of Ku80, Ku70 and NF90 to the IL-2 proximal promoter in vivo correlate with chromatin remodeling and transcriptional initiation in activated T-cells.


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