TY - JOUR
T1 - Immuno-electron microscopic identification of human estrogen receptor-DNA complexes at the estrogen-responsive element and in the first intron of a Xenopus vitellogenin gene
AU - ten Heggeler-Bordier, Béatrice
AU - Claret, François Xavier
AU - Wahli, Walter
N1 - Funding Information:
We thank J. Dubochet and R. Wittek for a critical reading of the manuscript. F.-X. Claret was the recipient of a France-Switzerland exchange fellowship. This work was supported by the Swiss National Science Foundation and the Etat de Vaud.
PY - 1988/11/5
Y1 - 1988/11/5
N2 - Using an extract of nuclei from the estrogen-responsive human breast cancer cell line MCF-7, protein-DNA complexes were assembled in vitro at the 5′ end of the Xenopus laevis vitellogenin gene B2 that is normally expressed in liver after estrogen induction. The complexes formed were analyzed by electron microscopy after labeling by the indirect colloidal gold immunological method using a monoclonal antibody specific for the human estrogen receptor. As identified by its interaction with protein A-gold, the antibody was found linked to two protein-DNA complexes, the first localized at the estrogen responsive element of the gene and the second in intron I, thus proving a direct participation of the receptor in these two complexes. The procedure used allows the visualization and rapid localization of specific transcription factors bound in vitro to a promoter or any other gene region.
AB - Using an extract of nuclei from the estrogen-responsive human breast cancer cell line MCF-7, protein-DNA complexes were assembled in vitro at the 5′ end of the Xenopus laevis vitellogenin gene B2 that is normally expressed in liver after estrogen induction. The complexes formed were analyzed by electron microscopy after labeling by the indirect colloidal gold immunological method using a monoclonal antibody specific for the human estrogen receptor. As identified by its interaction with protein A-gold, the antibody was found linked to two protein-DNA complexes, the first localized at the estrogen responsive element of the gene and the second in intron I, thus proving a direct participation of the receptor in these two complexes. The procedure used allows the visualization and rapid localization of specific transcription factors bound in vitro to a promoter or any other gene region.
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U2 - 10.1016/0022-2836(88)90612-2
DO - 10.1016/0022-2836(88)90612-2
M3 - Article
C2 - 3216393
AN - SCOPUS:0024235991
SN - 0022-2836
VL - 204
SP - 217
EP - 220
JO - Journal of Molecular Biology
JF - Journal of Molecular Biology
IS - 1
ER -