Transcription factors Ets1, NF-kappa B, and Sp1 are major determinants of the promoter activity of the human protein kinase CK2 alpha gene

CK2 alpha is one of two isoforms of protein kinase CK2, a highly conserved, ubiquitous, and vital phosphotransferase whose expression is kept at const ant cellular levels and whose dysregulated expression has been linked to ma lignant diseases. The upstream sequence of the gene coding for human CK2 al pha (CSNK1A1, chromosomal location 20p13) has been examined for promoter lo cation and transcription factor interactions using reporter gene assays (lu ciferase; HeLa cells), site-directed mutagenesis, electrophoretic mobility shift assays, super-shifts, UV cross-linking, Western blotting, and DNA aff inity chromatography. Highest promoter activity has been found in a region comprising positions -9 to 46, Factors Spl, Ets-1, and NF-kappa B have been identified as interaction partners and, by mutation of individual sites an d simultaneous mutations of two or more sites, shown to cross-talk to each other. At least two of the factors (Spl; NF-kappa B) were susceptible to ph osphorylation by CK2 holoenzyme, a tetramer: composed of two CK2 alpha and two regulatory CK2 beta proteins, but not by individual CK2 alpha, Because the phosphorylation decreases promoter binding and repeated immunoprecipita tion reveals presence of "free" CK2 beta in cell extracts, it is tempting t o speculate that the gene product CK2 alpha might readily form CK2 holoenzy me and feed back onto gene transcription. The data represent the first prom oter control analysis of a mammalian CK2 alpha gene and provide a hypothesi s of how the constant expression level of CK2 alpha may be achieved.