Correlation of nuclear acceptor sites for oestrogen receptors with gene transcription in vitro.
The interaction of oestrogen receptors with their nuclear acceptor sites was studied to ascertain whether these acceptor sites are involved in the regulation of ovalbumin-gene expression in the chick oviduct. As previously described, two distinct oestrogen-receptor species exist, and both are translocated into the nucleus after oestrogen administration in vivo [Smith, Clarke, Zalta & Taylor (1979) J. Steroid Biochem.10, 31-35]. In the present investigation we observed that the tubular-gland-cell concentrations of cytoplasmic receptors (800-900/cell) do not vary with prolonged withdrawal, nor do the relative ratios of the two receptor types change; however, the nuclear accumulation and retention of these receptors after secondary oestrogen administration are attenuated in a time-dependent fashion. Chicks were withdrawn from oestrogen for 24-84h. Some animals were then restimulated with oestrogen and killed after 4h, when oviduct nuclei were isolated. These nuclei were assayed for nuclear receptor concentrations and for their capacity to synthesize ovalbumin mRNA in vitro. Although an equal number of cytoplasmic receptors appeared to be translocated, oestrogen-receptor occupancy within the nucleus was not equal, but was inversely proportional to the preceding length of withdrawal. This decrease in nuclear acceptor sites was accompanied by a similar decrease in the capacity of these same nuclei to transcribe ovalbumin mRNA in vitro. A statistical evaluation of nuclear oestrogen-receptor concentrations and ovalbumin-mRNA synthesis in vitro was made. Correlation analysis revealed a Pearson coefficient r=0.87 (P<0.001, n=17), indicating that a high degree of correlation exists between these two parameters. These results are consistent with the hypothesis that nuclear oestrogen-receptor-acceptor complexes may correspond to initiation sites for RNA polymerase II transcription of an oestrogen-regulated gene.
D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism*; D002645:Chickens; D004054:Diethylstilbestrol / Q000494:pharmacology; D005260:Female; D066298:In Vitro Techniques; D007700:Kinetics; D010047:Ovalbumin / Q000235:genetics; D010057:Oviducts / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D011960:Receptors, Estrogen / Q000235:genetics* / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects; D014178:Translocation, Genetic / Q000187:drug effects
D012333:RNA, Messenger; D011960:Receptors, Estrogen; D004054:Diethylstilbestrol; D010047:Ovalbumin