NUCLEOTIDE STRUCTURE AND CHARACTERIZATION OF THE MURINE GENE ENCODINGANTICOAGULANT PROTEIN-C

The 15,160 bp murine gene encoding anticoagulation protein C (PC) was cloned and sequenced, including 414 bp upstream of exon 1 and 80 bp do wnstream of the translation stop codon. Nine exons and eight introns w ere identified. The first exon was untranslated and contained the majo r transcriptional start site, the surrounding nucleotide sequence of w hich matched reasonably well with the consensus eukaryotic Cap element sequence. The translational initiator methionine residue was located in exon 2. The other introns were positioned as splices between the ma jor domain units of the protein. The 5' untranslated region contained two possible CCAAT sequences and GC boxes, but no TATA box was obvious within the optimal range of distances from the transcription start si te. The 3'-flanking nucleotides included a probable polyadenylation si te (ATTAAA), beginning 80 nucleotides downstream of the translation st op codon, and a downstream consensus sequence (AGTGTTTC) required for the efficient formation of a 3' terminus of mRNA. Several high probabi lity transcription factor recognition sequences, including proteins th at are enriched in, or specific to, the liver, such as C/EBP alpha, C/ EBP beta, HNF1, and HNF3 beta, have been located in the 5' region of t he gene. These results indicate that all elements are present for live r-based transcription of the gene for murine PC.