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Molecular and Cellular Biology, December 2001, p. 7883-7891, Vol. 21, No. 23
0270-7306/01/$04.00+0   DOI: 10.1128/MCB.21.23.7883-7891.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Repression of Ets-2-Induced Transactivation of the Tau Interferon Promoter by Oct-4

Toshihiko Ezashi, Debjani Ghosh, and R. Michael Roberts*

Departments of Animal Sciences and Biochemistry, University of Missouri, Columbia, Missouri 65211

Received 30 January 2001/Returned for modification 12 March 2001/Accepted 23 August 2001

Oct-4 is a POU family transcription factor associated with potentially totipotent cells. Genes expressed in the trophectoderm but not in embryos prior to blastocyst formation may be targets for silencing by Oct-4. Here, we have tested this hypothesis with the tau interferon genes (IFNT genes), which are expressed exclusively in the trophectoderm of bovine embryos. IFNT promoters contain an Ets-2 enhancer, located at -79 to -70, and are up-regulated about 20-fold by the overexpression of Ets-2 in human JAr choriocarcinoma cells, which are permissive for IFNT expression. This enhancement was reversed in a dose-dependent manner by coexpression of Oct-4 but not either Oct-1 or Oct-2. When cells were transfected with truncated bovine IFNT promoters designed to eliminate potential octamer sites sequentially, luciferase reporter expression from each construct was still silenced by Oct-4. Full repression required both the N-terminal and POU domains of Oct-4, but neither domain used alone was an effective silencer. Oct-4 and Ets-2 formed a complex in vitro in the absence of DNA through binding of the POU domain of Oct-4 to a site located between the "pointed" and DNA binding domains of Ets-2. The two transcription factors were also coimmunoprecipitated after being expressed together in JAr cells. Oct-4, therefore, silences IFNT promoters by quenching Ets-2 transactivation. The POU domain most probably binds to Ets-2 directly, while the N-terminal domain inhibits transcription. These findings provide further evidence that the developmental switch to the trophectoderm is accompanied by the loss of Oct-4 silencing of key genes.

* Corresponding author. Mailing address: 158 Animal Science Research Center, University of Missouri, Columbia, MO 65211-5300. Phone: (573) 882-0908. Fax: (573) 882-6827. E-mail: robertsrm{at}missouri.edu.

Molecular and Cellular Biology, December 2001, p. 7883-7891, Vol. 21, No. 23
0270-7306/01/$04.00+0   DOI: 10.1128/MCB.21.23.7883-7891.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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