© 2003 by Society
© 2003 by the Society forNeuro-Oncology
Molecular cloning and identification of the human interleukin 13 alpha 2receptor (IL-13Ra2) promoter
Department of Neurosurgery (A.W., W.C.L.) and Graduate Program in Neuroscience (W.C.L.), University ofMinnesota Medical School, Minneapolis, MN 55455, USA, and Department of Neurosurgery, First Clinical College of ChinaMedical University, Shenyang, China (A.W.)
2 Address correspondence to Walter C. Low, Department of Neurosurgery,University of Minnesota Medical School, 2001 Sixth Street, SE, Minneapolis, MN55455, USA(lowwalt{at}umn.edu).
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Abstract |
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The interleukin 13 alpha 2 receptor (IL-13Ra2) has been shown to beexpressed in most malignant glioblastoma cells. Recent studies suggest thatIL-13Ra2 serves as a dominant negative inhibitor or a decoy receptor forIL-13. To investigate the transcriptional regulation of this receptor, wecloned and characterized the promoter for the human IL-13Ra2 gene.Our results demonstrate that this promoter contains three TATA boxes and oneCCAAT site. Several putative transcriptional factor binding sites for nuclearfactor of activated T cells 1, AP1 (c-JUN and c-FOS), AP2, GABP, OCT1, GATA3,PRE, and C-ETS1 were predicted in the promoter region. Using the secretedalkaline phosphate reporter gene assay, we investigated the functionalactivity of the human IL-13Ra2 promoter by transient transfection inglioma cell lines U118, U87, and T98, which differ in their expression of thehuman IL-13Ra2 protein. The different secreted alkaline phosphate activitiesamong these 3 cell lines suggest that the expression of humanIL-13Ra2 is regulated at the transcriptional level. Methylationanalysis showed that expression of IL-13Ra2 may not be the result ofmethylation of the CpG dinucleotides in the promoter region of the gene.Deletion analysis identified a 64 base pair (bp) region that is necessary forhuman IL-13Ra2 promoter activity. This 64-bp sequence containscis-elements for AP1, nuclear factor of activated T cells, and AP2. Thepossible role of AP1 in the regulation of human IL-13Ra2 promoteractivity was suggested by in vitro mutagenesis and c-JUN N-terminal kinaseinhibition analysis.
Received November 22, 2002; Accepted January 16, 2003
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