Analysis of the murine phosphoinositide 3-kinase gamma gene

Phosphoinositide 3-kinase gamma is preferentially expressed in leukocytes. PI3K gamma is activated by beta gamma subunits of heterotrimeric G-proteins , which thus link seven transmembrane helix receptor activation to phosphat idylinositol ( 3,4,5)-trisphosphate production. Here we describe the molecu lar cloning of the murine PI3K gamma cDNA, the PI3K gamma gene structure, i ts chromosomal assignment and the analysis of promoter activity. The mouse cDNA shares 86% identity to its pig and human orthologues at the nucleotide level. The MmPI3K gamma gene spans approximately 30 kb and comprises 11 ex ons. RACE-PCR indicated the presence of multiple start sites generating 5' UTRs with different lengths, the longest being 874 bp. The putative promote r region contains no TATA box but several putative binding sites for hemato poietic specific transcription factors. A 1200 bp long sequence upstream th e first transcription start site was found to possess tissue specific promo ter activity. Deletion constructs revealed two contiguous regions, with act ivator function, ranging from positions -139 to -557, and with inhibitory f unction, ranging from positions -557 to -892. FISH analysis revealed that t he MmPI3K gamma is located on chromosome 12 band B and that the human ortho logue is positioned on chromosome 7q22.2-22.3. In spite of some differences in the ATP-binding site, recombinant murine PI3K gamma protein is equally sensitive to wortmannin as its human counterpart. This suggests that mouse models will provide reliable results in the assessments of novel PI3K gamma inhibitors. (C) 2000 Elsevier Science B.V. All rights reserved.