Characterization of promoter region and genomic structure of the murine and human genes encoding Src like adapter protein

Src-like adapter protein (SLAP) was identified as a signaling molecule in a yeast two-hybrid system using the cytoplasmic domain of EphA2, a receptor protein tyrosine kinase (Pandey et al., 1995. Characterization of a novel S rc-like adapter protein that associates with the Eck receptor tyrosine kina se. J. Biol. Chem. 270, 19201-19204). It is very similar to members of the Src family of cytoplasmic tyrosine kinases in that it contains very homolog ous SH3 and SH2 domains (Abram and Courtneidge, 2000. Src family tyrosine k inases and growth factor signaling. Exp. Cell. Res. 254, 1-13.). However, i nstead of a kinase domain at the C-terminus, it contains a unique C-termina l region. In order to exclude the possibility that an alternative form exis ts, we have isolated genomic clones containing the murine Slap gene as well as the human SLA gene. The coding regions of murine Slap and human SLA gen es contain seven exons and six introns. Absence of any kinase domain in the genomic region confirm its designation as an adapter protein. Additionally , we have cloned and sequenced approximately 2.6 kb of the region 5' to the initiator methionine of the murine Slap gene. When subcloned upstream of a luciferase gene, this fragment increased the transcriptional activity abou t 6-fold in a human Jurkat T cell line and approximately 52-fold in a murin e T cell line indicating that this region contains promoter elements that d ictate SLAP expression. We have also cloned the promoter region of the huma n SLA gene. Since SLAP is transcriptionally regulated by retinoic acid and by activation of B cells, the cloning of its promoter region will permit a detailed analysis of the elements required for its transcriptional regulati on. (C) 2001 Elsevier Science B.V. All rights reserved.