Sequence of a novel mRNA coding for a C-terminal-truncated form of human NAD +-dependent 15-hydroxyprostaglandin dehydrogenase

• Published in: Gene Vol. 162; no. 2; pp. 319 - 322
• Main Authors: Pichaud, F., Frendo, J.L., Delage-Mourroux, R., de Vernejoul, M.C., Moukhtar, M.S., Jullienne, A.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 11.09.1995
• Subjects: 15-PGDH; ADH; alcohol dehydrogenase(s); Amino Acid Sequence; amino acid(s); base pair(s); Base Sequence; Cloning, Molecular; deoxyribonucleotide triphosphate; DNA, Complementary - genetics; dNTP; gene (DNA, mRNA) encoding 15-PGDH; Gene Expression; HL-60 cell line; HL-60 Cells; Humans; Hydroxyprostaglandin Dehydrogenases - genetics; Isoenzymes - genetics; kilobase(s) or 1000 bp; Molecular Sequence Data; Moloney murine leukemia virus; MoMLV; NAD +-dependent 15-hydroxyprostaglandin dehydrogenase; Northern blot; nucleic sequence; nucleotide(s); oligo; oligodeoxyribonucleotide; open reading frame; ORF; PCR; polymerase chain reaction; reverse transcription; RNA, Messenger - genetics; Sequence Alignment; Sequence Homology, Amino Acid; Sequence Homology, Nucleic Acid; type-I 15-PGDH; Western blot; Western blot; ORF; RT; gene (DNA, mRNA) encoding 15-PGDH; ADH; nt; dNTP; kilobase(s) or 1000 bp; open reading frame; type-I 15-PGDH; bp; reverse transcription; Northern blot; NAD +-dependent 15-hydroxyprostaglandin dehydrogenase; nucleotide(s); kb; oligo; base pair(s); polymerase chain reaction; aa; MoMLV; 15-PGDH; amino acid(s); deoxyribonucleotide triphosphate; oligodeoxyribonucleotide; alcohol dehydrogenase(s); HL-60 cell line; nucleic sequence; PCR; Moloney murine leukemia virus
• ISSN: 0378-1119; 1879-0038
• DOI: 10.1016/0378-1119(95)00319-2

We amplified, using the polymerase chain reaction (PCR) and NAD +-dependent 15-hydroxyprostaglandin dehydrogenase (type-I 15-PGDH)-specific primers, RNA extracted from the HL-60 cell line. Two bands, differing in size by approx. 160 bp, were detected with ethidium bromide staining after electrophoresis of amplification products and hybridization with a 15-PGDH-specific probe. Sequencing these DNA bands revealed that the largest corresponded to the 15-PGDH cloned from human placenta [Ensor et al., J. Biol. Chem. 265 (1990) 14888–14891]. The smaller sequence coded for a predicted C-terminal-truncated form of 15-PGDH. This subtype of the type-I 15-PGDH mRNA was also found using RT-PCR in human liver, placenta and a cell line derived from a human medullary thyroid carcinoma (TT cells). Hybridization studies using specific probes indicated that this new mRNA form probably corresponded to the 3.4-kb mRNA, one of the two 15-PGDH mRNAs previously detected in Northern blot analysis.