Estrogen sulfotransferase of the rat liver: complementary DNA cloning and age- and sex-specific regulation of messenger RNA.
Mammalian estrogen sulfotransferase (EST; EC 2.8.2.4) sulfurylates the hydroxyl group of estrogenic steroids by transferring the sulfate from a cosubstrate adenosine 3'-phosphate-5'-phosphosulfate. Sulfurylated steroids do not bind to the estrogen receptor with high affinity and, therefore, are hormonally inactive. We have purified rat liver EST and developed monoclonal antibody to this enzyme. By immunoscreening a lambda gt-11 expression library constructed from male rat liver cDNAs, the cDNA clone corresponding to EST was identified and isolated. A recombinant expression plasmid (pCMV5) containing this cDNA insert when transfected into COS-7 cells generated both immunologically and enzymatically active EST. With the help of this cDNA probe, we have explored the regulation of the EST mRNA in the liver and the possible role of this enzyme in sex hormone action. During the lifespan of male rats, only the young adult animals show hepatic androgen responsiveness. Also, estrogenic hormones strongly antagonize androgen action in the rat liver. Northern blot analysis of liver RNA derived from male rats of different ages shows that the androgen sensitivity of young adult animals is associated with a high expression of EST mRNA. During the same period, mRNA corresponding to dehydroepiandrosterone sulfotransferase is markedly (approximately 10-fold) down-regulated. Such a correlation is in concordance with the role of these enzymes in the maintenance of hepatic androgen sensitivity during young adult life by inactivating the estrogenic and sparing the androgenic steroids. Furthermore, the increase in the hepatic androgen sensitivity of androgen-treated female rats is also associated with the induction of EST.(ABSTRACT TRUNCATED AT 250 WORDS)
Demyan
W F
WF
Department of Cellular and Structural Biology, University of Texas Health Science Center, San Antonio 78284.
Song
C S
CS
Kim
D S
DS
Her
S
S
Gallwitz
W
W
Rao
T R
TR
Slomczynska
M
M
Chatterjee
B
B
Roy
A K
AK
D000367:Age Factors; D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D001483:Base Sequence; D015153:Blotting, Western; D002460:Cell Line; D003001:Cloning, Molecular; D003600:Cytosol / Q000201:enzymology; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D013196:Dihydrotestosterone / Q000494:pharmacology; D005260:Female; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D007527:Isoenzymes / Q000235:genetics; D008099:Liver / Q000201:enzymology* / Q000254:growth & development; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C / Q000276:immunology; D008969:Molecular Sequence Data; D010052:Ovariectomy; D010957:Plasmids; D011061:Poly A / Q000235:genetics; D012313:RNA / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D051381:Rats; D012689:Sequence Homology, Nucleic Acid; D012727:Sex Characteristics; D015238:Sulfotransferases / Q000235:genetics; D013466:Sulfurtransferases / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D014162:Transfection
D000911:Antibodies, Monoclonal; D007527:Isoenzymes; D012333:RNA, Messenger; D013196:Dihydrotestosterone; D011061:Poly A; D012313:RNA; D004247:DNA; D013466:Sulfurtransferases; D015238:Sulfotransferases; C019998:dehydroepiandrosterone sulfotransferase; C020122:estrone sulfotransferase