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Volume 271, Number 28, Issue of July 12, 1996 pp. 16683-16689
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.

Silencing of the Gene for the beta  Subunit of Human Chorionic Gonadotropin by the Embryonic Transcription Factor Oct-3/4

(Received for publication, February 21, 1996, and in revised form, April 25, 1996)

Limin Liu Dagger and R. Michael Roberts §

From the Departments of Dagger  Biological Sciences and § Animal Sciences and Biochemistry, University of Missouri, Columbia, Missouri 65211

The transcription factor Oct-3/4 may be important in maintaining embryonic cells in an undifferentiated state. It is probably down-regulated at about the time that human chorionic gonadotropin (hCG) is first expressed in embryonic trophectoderm. Here we report that Oct-3/4 strongly inhibits the hCGbeta subunit (hCGbeta ) promoter in JAr choriocarcinoma cells. Oct-3/4 reduced chloramphenicol acetyltransferase (CAT) reporter expression from the -305hCGbeta promoter by about 90% in transient co-transfection assays, but had no effect on expression from the -249hCGbeta promoter. The -305/-249 hCGbeta fragment specifically bound synthetic Oct-3/4 protein as measured in electrophoretic mobility shift assays, and the Oct-3/4-binding site was localized around -270 by methylation interference footprinting. Site-directed mutagenesis of this binding site abolished Oct-3/4 repression. When stably transfected into JAr cells, Oct-3/4 reduced the amounts of both endogenous hCGbeta messenger RNA and hCG protein to less than 10% of controls. We suggest that silencing of Oct-3/4 in trophectoderm is a prerequisite for hCG up-regulation in early human embryos at the time of maternal recognition of pregnancy.


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