4D (PDUD), part of the complex CAMP-specific PDE4 family, plays a pivotal role in the regulation of airway smooth muscle relaxation. Its gene on 5q12 encodes 5 splice-variants which show tissue-dependent expression and regulation. We have demonstrated both transcriptional and functional activity upregulation of PDE4D5 in human airway smooth muscle cells (hASM) in response to increased intracellular CAMP. In silico methods were used to determine genomic arrangement and identify a putative promoter of PDE4D5. 1544 and 621 b.p. constructs of this putative promoter were ligated into the luciferase reporter vector PGL3 enhancer, and transiently transfected into a CHO cell line that stably expressed both a2 adrenoceptors and a secreted placental alkaline phosphatase (SPAP) reporter driven by a CAMP responsive promoter. The cells were then stimulated for 5 hours with isoprenaline, isoprenaline and IBMX (non-selective PDE inhibitor), lBMX alone, and forskolin at doses calculated to produce a maximal CAMP increase. SPAP output was increased by up to 4 fold from basal (p
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