Regulation of choline kinase R: analyses of alternatively spliced choline kinases and the promoter region.

The previous report described the cloning kinase R1 cDNA [Uchida, T. and Yamashita, S. (1992) J. Biol. Chem. 267, 10156-10162]. A new cDNA, for choline kinase R2, was isolated from rat liver cDNA libraries. This transcript was thought to be generated by alternative splicing. Ribonuclease protection analysis revealed the presence of a third choline kinase. Three transcripts were detected in all of the examined rat tissues at different levels. Southern blot analysis demonstrated a single copy of choline kinase R gene. The genomic DNA containing the first exon and its flanking regions of choline kinase R gene was isolated and characterized. A number of transcription start sites, determined by ribonuclease protection and primer extension analyses, were found. The most 3' site, 193 base pairs upstream of the initiation codon and common in liver and testis, was the main site in liver. Some transcription start sites were detected only in testis. Choline kinase R gene showed features not only of a typical housekeeping gene but also of a gene regulated through a variety of putative cis-acting motifs. 3-Methylcholanthrene and carbon tetrachloride increased all of three transcripts to various levels, and enhanced transcription from the same start site, which scarcely gave a detectable product in normal liver.