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Transcriptional Activation of the SH2D2A Gene Is Dependent on a Cyclic Adenosine 5'-Monophosphate-Responsive Element in the Proximal SH2D2A Promoter¹

Ke-Zheng Dai^*,, Finn-Eirik Johansen, Kristin Melkevik Kolltveit, Hans-Christian Aasheim^¶, Zlatko Dembic, Frode Vartdal^* and Anne Spurkland^2,*,

^* Institute of Immunology and Institute of Pathology, National University Hospital; Department of Oral Biology and Institute of Anatomy, University of Oslo; and ^¶ Department of Immunology, Norwegian Radium Hospital, Oslo, Norway

The SH2D2A gene, encoding the T cell-specific adapter protein (TSAd), is rapidly induced in activated T cells. In this study we investigate the regulation of the SH2D2A gene in Jurkat T cells and in primary T cells. Reporter gene assays demonstrated that the proximal 1-kb SH2D2A promoter was constitutively active in Jurkat TAg T cells and, to a lesser extent, in K562 myeloid cells, Reh B cells, and 293T fibroblast cells. The minimal SH2D2A promoter was located between position –236 and –93 bp from the first coding ATG, and transcriptional activity in primary T cells depended on a cAMP response element (CRE) centered around position –117. Nuclear extracts from Jurkat TAg cells and activated primary T cells contained binding activity to this CRE, as observed in an EMSA. Consistent with this observation, we found that a cAMP analog was a very potent inducer of SH2D2A mRNA expression in primary T cells as measured by real-time RT-PCR. Furthermore, activation of SH2D2A expression by CD3 stimulation required cAMP-dependent protein kinase activity. Thus, transcriptional regulation of the SH2D2A gene in activated T cells is critically dependent on a CRE in the proximal promoter region.

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