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- 저자 : James D. Godkin (검색결과 3 건)
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Min-Su Kim,Toshihiro Sakurai,Daisuke Sato,Kyu-Tae Chang,Kwan-Sik Min,James D. Godkin,Kazuhiko Imakawa 한국동물번식학회 2012 Reproductive & Developmental Biology(Supplement) Vol.36 No.2s
Interferon-tau (IFNT) is regarded, generally, to be the conceptus protein that signals maternal recognition of pregnancy in ruminants. Although the discovery was made over two decades ago, the molecular mechanisms that regulate IFNT expression are not well understood. Previous studies demonstrated that transcription factors, caudal-related homeobox- 2 (CDX2), JUN, ETS2 and a transcriptional coactivator CREB binding protein (CREBBP) positively influenced IFNT gene expression, while OCT4 may exhibit negative regulation. We and others have observed that both CDX2 and OCT4 coexist during early stages of conceptus elongation but as development proceeds, OCT4 expression diminishes. The objective of this study was to evaluate the stimulatory and inhibitory effects of CDX2 and OCT4, respectively, on IFNT gene transcription when evaluated with other transcription factors. Human choriocarcinoma JEG3 cells were co-transfected with an ovine IFNT (-654 base pair)-luciferase reporter (-654-oIFNT-Luc) construct and several transcription factor expression plasmids. When the reporter construct was co-transfected with either CDX2, ETS2 or CJUN, transcription of -654-oIFNT-Luc increased about two-fold compared to -654-oIFNT-Luc alone. When -654-oIFNT-Luc was co-transfected with both c-jun and Ets-2, activity of -654-oIFNT-Luc was increased about four-fold; cotransfection with JUN, ETS2 and CDX2 increased -654-oIFNT-Luc expression 12X, indicating that the stimulatory activity of the transcription factors was additive. OCT4, when cotransfected with -654-oIFNT-Luc, reduced expression of the later about 40% when compared to -654-oIFNT-Luc alone. When co-transfected with JUN and/or ETS2, OCT4 abolished the stimulatory effect of these transcription factors. OCT4 also inhibited the stimulatory activity of CDX2 alone, but not when CDX2 was combined with JUN and/or ETS2. Therefore, when combined with the other transcription factors, CDX2 over the transcriptional inhibitory activity of OCT4. Conversely, when cells were transfected initially with OCT4 (0h) followed by transfection with CDX2, ETS2 and JUN 24 h later, -654-oIFNT-Luc expression was reduced to control (-654-oIFNT-Luc, alone) levels. Not surprisingly, 12S E1A, an inhibitor of transcriptional coactivator CREBBP, reduced stimulation of -654- oIFNT-Luc expression by CDX2, ETS2 and JUN, in combination, by about 40%, indicating that proper transcription complex formation is required for maximum expression. In conclusion, it is suggested that prior to conceptus elongation, pre-existing OCT4 may inhibit IFNT expression, but as elongation proceeds, IFNT expression increases, resulting from incremental increases in CDX2 expression, diminished OCT4 expression, and possibly proper transcription factor complex formation. Key words) Interferon-tau, CDX2, OCT4, transcription
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Kim, Min-Su,Sakurai, Toshihiro,Bai, Hanako,Bai, Rulan,Sato, Daisuke,Nagaoka, Kentaro,Chang, Kyu-Tae,Godkin, James D.,Min, Kwan-Sik,Imakawa, Kazuhiko Asian Australasian Association of Animal Productio 2013 Animal Bioscience Vol.26 No.5
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.
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