Presence of Transcription Factor OCT4 Limits Interferon-tau Expression during the Pre-attachment Period in Sheep

  • Kim, Min-Su (Laboratory of Animal Breeding, Graduate School of Agricultural and Life Sciences, The University of Tokyo) ;
  • Sakurai, Toshihiro (Laboratory of Animal Breeding, Graduate School of Agricultural and Life Sciences, The University of Tokyo) ;
  • Bai, Hanako (Laboratory of Animal Breeding, Graduate School of Agricultural and Life Sciences, The University of Tokyo) ;
  • Bai, Rulan (Laboratory of Animal Breeding, Graduate School of Agricultural and Life Sciences, The University of Tokyo) ;
  • Sato, Daisuke (Laboratory of Animal Breeding, Graduate School of Agricultural and Life Sciences, The University of Tokyo) ;
  • Nagaoka, Kentaro (Laboratory of Veterinary Physiology, Tokyo University of Agriculture and Technology) ;
  • Chang, Kyu-Tae (Primate Research Center, Korea Research Institute of Biosciences and Biotechnologies) ;
  • Godkin, James D. (Department of Animal Science, University of Tennessee) ;
  • Min, Kwan-Sik (Animal Biotechnology, Hankyong National University) ;
  • Imakawa, Kazuhiko (Laboratory of Animal Breeding, Graduate School of Agricultural and Life Sciences, The University of Tokyo)
  • Received : 2012.08.29
  • Accepted : 2012.10.18
  • Published : 2013.05.01


Interferon-tau (IFNT) is thought to be the conceptus protein that signals maternal recognition of pregnancy in ruminants. We and others have observed that OCT4 expression persists in the trophectoderm of ruminants; thus, both CDX2 and OCT4 coexist during the early stages of conceptus development. The aim of this study was to examine the effect of CDX2 and OCT4 on IFNT gene transcription when evaluated with other transcription factors. Human choriocarcinoma JEG-3 cells were cotransfected with an ovine IFNT (-654-bp)-luciferase reporter (-654-IFNT-Luc) construct and several transcription factor expression plasmids. Cotransfection of the reporter construct with Cdx2, Ets2 and Jun increased transcription of -654-IFNT-Luc by about 12-fold compared with transfection of the construct alone. When cells were initially transfected with Oct4 (0 h) followed by transfection with Cdx2, Ets2 and/or Jun 24 h later, the expression of -654-IFNT-Luc was reduced to control levels. OCT4 also inhibited the stimulatory activity of CDX2 alone, but not when CDX2 was combined with JUN and/or ETS2. Thus, when combined with the other transcription factors, OCT4 exhibited little inhibitory activity towards CDX2. An inhibitor of the transcriptional coactivator CREB binding protein (CREBBP), 12S E1A, reduced CDX2/ETS2/JUN stimulated -654-IFNT-Luc expression by about 40%, indicating that the formation of an appropriate transcription factor complex is required for maximum expression. In conclusion, the presence of OCT4 may initially minimize IFNT expression; however, as elongation proceeds, the increasing expression of CDX2 and formation of the transcription complex leads to greatly increased IFNT expression, resulting in pregnancy establishment in ruminants.


  1. Roberts, R. M., J. C. Cross and D. W. Leaman. 1992. Interferons as hormones of pregnancy. Endocr. Rev. 13:432-452.
  2. Ralston, A. and J. Rossant. 2005. Genetic regulation of stem cell origins in the mouse embryo. Clin. Genet. 68:106-112.
  3. Saadeldin, I. M., W. Choi, B. R. da Torre, B. Kim, B. Lee and G. Jang. 2012. Embryonic development and implantation related gene expression of oocyte reconstructed with bovine trophoblast cells. J. Reprod. Dev. 58:425-431.
  4. Sakurai, T., A. Sakamoto, Y. Muroi, H. Bai, K. Nagaoka, K. Tamura, T. Takahashi, K. Hashizume, M. Sakatani, M. Takahashi, J. D. Godkin and K. Imakawa. 2009. Induction of endogenous interferon tau gene transcription by CDX2 and high acetylation in bovine nontrophoblast cells. Biol. Reprod. 80:1223-1231.
  5. Sakurai, T., H. Bai, T. Konno, A. Ideta, Y. Aoyagi, J. D. Godkin and K. Imakawa. 2010. Function of a transcription factor CDX2 beyond its trophectodermal lienage specification. Endocrinology 151:5873-5881.
  6. Stewart, H. J., S. H. McCann, P. J. Barker, K. E. Lee, G. E. Lamming and A. P. Flint. 1987. Interferon sequence homology and receptor binding activity of ovine trophoblast antiluteolytic protein. J. Endocrinol. 115:R13-R15.
  7. Suh, E., L. Chen, J. Taylor and P. G. Traber. 1994. A homeodomain protein related to caudal regulates intestine-specific gene transcription. Mol. Cell. Biol. 14:7340-7351.
  8. Xu, N., Y. Takahashi, F. Matsuda, S. Sakai, R. K. Christenson and K. Imakawa. 2003. Coactivator CBP in the regulation of conceptus IFN gene transcription. Mol. Reprod. Dev. 65:23-29.
  9. Yamaguchi, H., Y. Ikeda, J. I. Moreno, M. Katsumura, T. Miyazawa, E. Takahashi, K. Imakawa, S. Sakai and R. K. Christenson. 1999. Identification of a functional transcriptional factor AP1 site in the sheep interferon-gene that mediates a response to PMA in JEG3 cells. Biochem. J. 340:767-773.
  10. Yamaguchi, H., K. Nagaoka, K. Imakawa, S. Sakai and R. K. Christenson. 2001. Enhancer regions of ovine interferon-gene that confer PMA response or cell type specific transcription. Mol. Cell. Endocrinol. 173:147-155.
  11. Ezashi, T., D. Ghosh and R. M. Roberts. 2001. Repression of Ets-2 induced transactivation of the interferon-promoter by Oct-4. Mol. Cell. Biol. 21:7883-7891.
  12. Farin, C. E., K. Imakawa and R. M. Roberts. 1989. In situ localization of mRNA for the interferon, ovine trophoblast protein-1, during early embryonic development of the sheep. Mol. Endocrinol. 3:1099-1107.
  13. Godkin, J. D., F. W. Bazer, J. Moffatt, F. Sessions and R. M. Roberts. 1982. Purification and properties of a major, low molecular weight protein relaeased by the trophoblast of sheep blastocysts at day 13-21. J. Reprod. Fertil. 65:141-150.
  14. Guillomot, M., C. Michel, P. Gaye, N. Charlier, J. Trojan and J. Martal. 1990. Cellular localization of an embryonic interferon, ovine trophoblastin and its mRNA in sheep embryos during early pregnancy. Biol. Cell 68:205-211.
  15. Imakawa, K., R. V. Anthony, M. Kazemi, K. R. Marotti, H. G. Polites and R. M. Roberts. 1987. Interferon-like sequence of ovine trophoblast protein secreted by embryonic trophectoderm. Nature 330:377-379.
  16. Imakawa, K., M. S. Kim, F. Matsuda-Minehata, S. Ishida, M. Iizuka, M. Suzuki, K. T. Chang, S. E. Echternkamp and R. K. Christenson. 2006. Regulation of the ovine interferon-tau gene by a blastocyst-specific transcription factor, Cdx2. Mol. Reprod. Dev. 73:559-567.
  17. Kurosaka, S., S. Eckardt and K. J. McLaughlin. 2004. Pluriopotent lineage definition in bovine embryos by Oct4 transcript localization. Biol. Reprod. 71:1578-1582.
  18. Matsuda, F., N. Xu, S. Kijima, C. Tachi, R. K. Christenson, S. Sakai and K. Imakawa. 2004. Analysis of transcriptional control elements in the 5'-upstream region of ovine interferon-gene using feeder-independent caprine trophoblast cell line, HTS-1. Placenta 25:166-175.
  19. Nagaoka, K., H. Nojima, F. Watanabe, K. T. Chang, R. K. Christenson, S. Sakai and K. Imakawa. 2003. Regulation of blastocyst migration, apposition, and initial adhesion by a chemokine, interferon -inducible protein 10 kDa (IP-10), during early gestation. J. Biol. Chem. 278:29048-29056.
  20. Nephew, K. P., A. E. Whaley, R. K. Christenson and K. Imakawa. 1993. Differential expression of distinct mRNAs for ovine trophoblast protein-1 and related sheep type I interferons. Biol. Reprod. 48:768-778.
  21. Nichols, J., B. Zevnik, K. Anastassiadis, H. Niwa, D. Klewe-Nebenius, I. Chambers, H. Scholer and A. Smith. 1998. Formation of pluripotent stem cells in the mammalian embryo depends on the POU transcription factor Oct4. Cell 95:379-391.
  22. Niwa, H., Y. Toyooka, D. Shimosato, D. Strumpf, K. Takahashi, R. Yagi and J. Rossant. 2005. Interaction between Oct3/4 and Cdx2 determines trophectoderm differentiation. Cell 123:917-929.
  23. Palmieri, S. L., W. Peter, H. Hess and H. R. Scholer. 1994. Oct-4 transcription factor is differentially expressed in the mouse embryo during establishment of the first two extraembryonic cell lineages involved in implantation. Dev. Biol. 166:259-267.
  24. Bannister, A. J. and T. Kouzarides. 1995. CBP-induced stimulation of c-Fos activity is abrogated by E1A. EMBO J. 14:4758-4762.
  25. Beck, F., T. Erler, A. Russell and R. James. 1995. Expression of Cdx-2 in the mouse embryo and placenta: possible role in patterning the extra-embryonic membranes. Dev. Dyn. 204:219-227.
  26. Berg, D. K., C. S. Smith, D. J. Pearton, D. N. Wells, R. Broadhurst, M. Donnison and P. L. Pfeffer. 2011. Trophectoderm lienage determination in cattle. Dev. Cell 20:244-255.
  27. Chawengsaksophak, K., R. James, V. E. Hammond, F. Kontgen and F. Beck. 1997. Homeosis and intestinal tumours in Cdx2 mutant mice. Nature 386:84-87.
  28. Degrelle, S. A., E. Campion, C. Cabau, F. Piumi, P. Reinaud, C. Richard, J. P. Renard and I. Hue. 2005. Molecular evidence for critical period in mural trophoblast development in bovine blastocysts. Dev. Biol. 288:448-460.
  29. Demmers, K. J., K. Derecka and A. Flint. 2001. Trophoblast interferon and pregnancy. Reproduction 121:41-49.
  30. Ezashi, T., A. D. Ealy, M. C. Ostrowski and R. M. Roberts. 1998. Control of interferon- gene expression by Ets2. Proc. Natl. Acad. Sci. USA 95:7882-7887.

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