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Department of Immunology, Institute of International Health, Immunology and Microbiology, University of Copenhagen, Copenhagen, Denmark
In this study, we characterize the molecular signal pathways that lead to MHC class I chain-related protein A (MICA) expression after histone deacetylase (HDAC)-inhibitor (HDAC-i) treatment of Jurkat T cells. Chelating calcium with BAPTA-AM or EGTA potently inhibited HDAC- and CMV-mediated MICA/B expression. It was further observed that endoplasmic reticulum calcium stores were depleted after HDAC treatment. NF-
B activity can be induced by HDAC treatment. However, nuclear translocation of NF-B p65 was not observed after HDAC treatment of Jurkat T cells and even though we could effectively inhibit p65 expression by siRNA, it did not modify MICA/B expression. To identify important elements in MICA regulation, we made a promoter construct consisting of 
3 kb of the proximal MICA promoter in front of GFP. Deletion analysis showed that a germinal center-box containing a putative Sp1 site from position –113 to –93 relative to the mRNA start site was important for HDAC and CMV-induced promoter activity. Sp1 was subsequently shown to be important, as targeted mutation of the Sp1 binding sequence or siRNA mediated down modulation of Sp1-inhibited MICA promoter activity and surface-expression.
The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
1 This study was supported from The Danish Medical Research Council, the Novo Nordisk Foundation, the Danish Cancer Research Foundation, the A.P. Møller Foundation for Advancement of Medical Science (to S.S.); L.A. was supported from the Copenhagen Cluster of Immunology. H.J. and M.T.P. were supported by the Novo Nordisk Foundation. K.A.H. was supported by the Danish Cancer Society.
2 L.A. and H.J. contributed equally to the work.
3 Current address: Biotech Research and Innovation Centre (BRIC), Centre for Epigenetics, University of Copenhagen, Copenhagen, Denmark.
4 Address correspondence and reprint requests to Dr. Søren Skov: Department of Immunology, Institute of International Health, Immunology and Microbiology, Building 6-6-14, Blegdamsvej 3, University of Copenhagen, Denmark. E-mail address: s.skov{at}immi.ku.dk
5 Abbreviations used in this paper: HDAC-i, histone deacetylase-inhibitors; ER, endoplasmic reticulum; WT, wild type; ChIP, chromatin immunoprecipitation; MICA, MHC class I chain-related protein A.
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