Close kinship of human 20alpha-hydroxysteroid dehydrogenase gene with three aldo-keto reductase genes.
BACKGROUND
20alpha-Hydroxysteroid dehydrogenase (HSD) is a member of the aldo-keto reductase (AKR) superfamily and catalyses the reaction of progesterone to the inactive form 20alpha-hydroxyprogesterone. Progesterone plays an important role in the maintenance of pregnancy, and, in rodents, plasma progesterone levels decrease abruptly just before parturition. The induction of 20alpha-HSD is thought to be responsible for the decrease in plasma progesterone at term. High homology between human 20alpha-HSD [AKR 1C1] cDNA with other AKRs had caused difficulty in gene isolation and expression analysis. Thus, the metabolism of progesterone in the human reproductive system remained unclear.


RESULTS
By hybridization with rat 20alpha-HSD [AKR 1C8] cDNA and high-stringency polymerase chain reaction (PCR) with gene-specific primers, we were able to isolate the human 20alpha-HSD, bile acid-binding protein (BABP) [AKR 1C2], prostaglandin F synthase (PGFS) [AKR 1C3], and dihydrodiol dehydrogenase (DD) 4 [AKR 1C4] genes. These genes had similar exon-intron organizations and shared a high homology. The four recombinant enzymes encoded by these genes showed distinct substrate specificity. By reverse transcription-PCR analysis, human 20alpha-HSD, BABP and PGFS mRNAs were expressed ubiquitously, while DD4 mRNA was restricted to the liver. Promoter activities of the 20alpha-HSD, BABP and PGFS genes were high, both in ovarian granulosa cells and hepatocytes. Radiation hybridization analysis revealed that all these genes were located close together in chromosome 10.


CONCLUSIONS
The human gene encoding for the progesterone-metabolizing enzyme 20alpha-HSD in the female reproductive system was cloned, and its expression and gene localization were elucidated. BABP, PGFS and DD4 genes, which were highly homologous to the 20alpha-HSD gene, were also cloned, and their structure and function were characterized.
Nishizawa
M
M
Department of Medical Chemistry; Department of Obstetrics and Gynecology, Kansai Medical University, 10-15 Fumizono, Moriguchi, Osaka 570-8506, Japan.
Nakajima
T
T
Yasuda
K
K
Kanzaki
H
H
Sasaguri
Y
Y
Watanabe
K
K
Ito
S
S
D015089:20-Hydroxysteroid Dehydrogenases / Q000235:genetics* / Q000378:metabolism*; D000818:Animals; D002352:Carrier Proteins / Q000235:genetics* / Q000378:metabolism; D002879:Chromosomes, Human, Pair 10; D003001:Cloning, Molecular; D005091:Exons; D005260:Female; D006801:Humans; D006911:Hydroxyprostaglandin Dehydrogenases / Q000235:genetics* / Q000378:metabolism; D006913:Hydroxysteroid Dehydrogenases; D007438:Introns; D007658:Ketone Oxidoreductases / Q000235:genetics / Q000378:metabolism; D008562:Membrane Glycoproteins; D008969:Molecular Sequence Data; D010088:Oxidoreductases / Q000235:genetics* / Q000378:metabolism; D010920:Placenta / Q000201:enzymology; D016133:Polymerase Chain Reaction / Q000379:methods; D011247:Pregnancy; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger; D051381:Rats; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012689:Sequence Homology, Nucleic Acid; D013379:Substrate Specificity; D014599:Uterus / Q000201:enzymology; D045002:3-alpha-Hydroxysteroid Dehydrogenase (B-Specific)
D015089:20-Hydroxysteroid Dehydrogenases; D002352:Carrier Proteins; D006911:Hydroxyprostaglandin Dehydrogenases; D006913:Hydroxysteroid Dehydrogenases; D007658:Ketone Oxidoreductases; D008562:Membrane Glycoproteins; D010088:Oxidoreductases; D012333:RNA, Messenger; D011994:Recombinant Proteins; C037643:bile acid binding proteins; C031051:prostaglandin-F synthase; C435496:AKR1C2 protein, human; D045002:3-alpha-Hydroxysteroid Dehydrogenase (B-Specific); C048338:trans-1,2-dihydrobenzene-1,2-diol dehydrogenase