Mechanism of estrogen receptor-dependent transcription in a cell-free system.
RNA synthesis was stimulated directly in a cell-free expression system by crude preparations of recombinant mouse estrogen receptor (ER). Receptor-stimulated transcription required the presence of estrogen response elements (EREs) in the test template and could be specifically inhibited by addition of competitor oligonucleotides containing EREs. Moreover, polyclonal antibodies directed against the DNA-binding region of ER inhibited ER-dependent transcription. In our cell-free expression system, hormone-free ER induced transcription in a hormone-independent manner. Evidence is presented suggesting that ER acts by facilitating the formation of a stable preinitiation complex at the target gene promoter and thus augments the initiation of transcription by RNA polymerase II. These observations lend support to our current understanding of the mechanism of steroid receptor-regulated gene expression and suggest strong conservation of function among members of the steroid receptor superfamily.
Elliston
J F
JF
Department of Cell Biology, Baylor College of Medicine, Houston, Texas 77030.
Fawell
S E
SE
Klein-Hitpass
L
L
Tsai
S Y
SY
Tsai
M J
MJ
Parker
M G
MG
O'Malley
B W
BW
D000818:Animals; D001483:Base Sequence; D002474:Cell-Free System; D004268:DNA-Binding Proteins / Q000378:metabolism*; D005822:Genetic Vectors; D007304:Insect Viruses / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D011401:Promoter Regions, Genetic; D011960:Receptors, Estrogen / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D016385:TATA Box; D013698:Templates, Genetic; D014158:Transcription, Genetic
D004268:DNA-Binding Proteins; D015345:Oligonucleotide Probes; D011960:Receptors, Estrogen; D011994:Recombinant Proteins