In vitro characterization of an estrogen-regulated mRNA stabilizing activity in the avian liver.
Estrogen-mediated accumulation of the avian apolipoprotein (apo) II mRNA is in part due to its stabilization. To identify the biochemical activity responsible for this effect, radiolabeled, capped, and polyadenylated apoII mRNA was incubated in vitro in liver cytosolic extracts from roosters who received either estrogen (estrogen-treated extract) or the vehicle (control extract) parenterally. The mRNA was very stable in estrogen-treated extract but was rapidly degraded in control extract. The RNA was degraded predominantly by endonuclease rather than exonuclease activity. The addition of the estrogen-treated extract to the control extract prevented the degradation of the mRNA in trans. This biochemical activity was heat labile and was also destroyed by proteinase K but not by micrococcal nuclease, indicating that estrogen treatment resulted in the expression of a protein in the liver that stabilized the apoII mRNA by inhibiting its nucleolytic degradation. This mRNA stabilization factor was labile around 60 degrees C, whereas the RNase remained stable up to 80 degrees C. Studies on mRNA protein interaction showed that both control and estrogen-treated extracts contain mRNA-binding (mRNP) proteins that bind apoII mRNA. An increased binding to apoII mRNA by a subset of these proteins was observed with estrogen-treated extract as compared with the control extract. This activity, although it afforded complete protection from nucleolytic degradation to apoII and apo A1 mRNAs, appeared to provide less protection to mRNAs encoding chicken serum albumin and vitellogenin, suggesting differential stabilization of mRNAs. These studies indicate that a cytosolic mRNA-stabilization factor, providing apoII mRNA complete protection from nucleolytic degradation, is expressed in the avian liver upon estrogen treatment. This appears to be the first time that a biochemical activity responsible for hormone-mediated stabilization of mRNAs and estrogen induction of mRNA binding by specific mRNPs have been identified and partially characterized in vitro.
Ratnasabapathy
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Department of Pharmacology, Arnold and Marie Schwartz College of Pharmacy, Long Island University, Brooklyn, NY 11201 USA.
D000818:Animals; D016632:Apolipoprotein A-I / Q000235:genetics; D001053:Apolipoproteins / Q000235:genetics* / Q000378:metabolism; D015152:Blotting, Northern; D002645:Chickens; D003600:Cytosol / Q000378:metabolism; D004967:Estrogens / Q000494:pharmacology*; D006358:Hot Temperature; D008099:Liver / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D008297:Male; D011498:Protein Precursors / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000378:metabolism*; D014819:Vitellogenins / Q000235:genetics
D016632:Apolipoprotein A-I; D001053:Apolipoproteins; D004967:Estrogens; D011498:Protein Precursors; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D014819:Vitellogenins; C054949:apolipoprotein II