Cloning and characterization of mouse NAD+-dependent 15-hydroxyprostaglandin dehydrogenase (15-PGDH) CDNA and genomic dna

15-PGDH is a key catabolic enzyme that controls the biological activities of prostaglandins. The cDNA for mouse 15-PGDH was cloned from a lung cDNA library. The cDNA contains a 798 bp open reading frame that codes for a protein of 266 amino acids (Mr 28,775) which shares 87% identity with the human 15-PGDH. The region that shows major difference in sequence resides in the C-terminal. The regions that are believed to form the NAD+ binding sites and the active site are conserved in mouse and human enzymes. The authenticity of the cDNA was confirmed by expression of an active 15-PGDH in E. coli. The specific activity of the mouse enzyme was comparable to that of the human enzyme. Northern blot analysis showed that mouse 15-PGDH mRNA was expressed primarily in lung, intestine, kidney and spleen. The entire mouse 15-PGDH gene and its S'-flanking region was cloned from a 129 mouse ES bacteriophage PI genomic library. The gene containing 4 exons is 5.3 Kb in length. Nucleotide seguence of 1.2 Kb promoter region contains a TATA box and a number of potential regulatory elements including Spl, ATF/CRE GRE, AP2 and NF-IL6.