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Regulation of the Bovine New Type I Interferon-tau Gene Transcription
Min-Su Kim,Kwan-Sik Min,Kazuhiko Imakawa,Ik Soo Jeon,Sung Woo Kim 한국동물생명공학회(구 한국동물번식학회) 2017 Reproductive & Developmental Biology(Supplement) Vol.41 No.2
Interferon-tau (IFNT) gene was originally recognized as as low molecular weight secretory protein produced by the conceptus during the peri-implantation period in ruminant ungulates. Two isoform interferon tau (IFNT) genes exist in bovine conceptus. An antiluteolytic substance secreted by the bovine conceptus and primarily responsible for maternal recognition of pregnancy is bovine trophoblast protein 1 (bIFNT1), a new type I bovine interferon tau (bIFNTc1) gene. However, studies of new type I bovine interferon- tau (bIFNTc1) gene regulation, an anti-luteolytic factor produced by conceptuses of the ruminant ungulates, have been carried out, but a definitive mechanism for its transcription has not been elucidated. In the present study, Human choriocarcinoma JEG3 cells were co-transfected with an bovine IFNTc1 (–2.2-kb, –631-bp)-luciferase reporters (–2.2-kb, –631-bp-Luc) construct and several transcription factor expression plasmids. bIFNTc1 gene was had very effect on activity by alone ETS2, and/or AP1 (JUN) expression factors in choriocarcinoma JEG3 cell. However, bIFNTc1 (–2.2-kb vs –631-bp)-luciferase reporters, expression level of the upstream region was not identified. The degree of transcriptional activation of the bIFNTc1(–2.2-kb)-luciferase reporter gene was had more effect by ETS2 and/or AP1, expression plasmid. These results indicate that an more ETS2 and/or AP1-binding site may be located between positions –2.2 kb to –631 bp of the bIFNTc1 gene. Also, transcription of the (–631-IFNT1c1)- Luc increased more than 30 times when the potential relationship between AP1 and ETS2, and their association with a co-activator, cAMP-response element binding protein- binding protein (CREBBP). These observation suggest that AP1 and ETS2 are the most probable binding partners for CREBBP in the potentiation of bIFNTc1 gene transcription. These results indicate that new type I bovine interferon tau (bIFNTc1) gene transcription is regulated by AP1 and ETS2, and suggest that AP1 and ETS2 could be a key molecule in determining bIFNTc1 gene transcription by the trophectoderm.
Kim, Min-Su,Min, Kwan-Sik,Imakawa, Kazuhiko Asian Australasian Association of Animal Productio 2013 Animal Bioscience Vol.26 No.6
Various endometrial genes in ruminant ungulates are regulated by conceptus interferon tau (IFNT). However, the effect of each IFNT isoform has not been carefully evaluated. In this study, the effects of 2 IFNT isoforms, paralogs found in utero, and interferon alpha (IFNA) on uterine epithelial and Mardin-Darby bovine kidney (MDBK) cells were evaluated. Expression vectors of the bovine interferon (bIFNT) genes bIFNT1, bIFNTc1, and bIFNA were constructed, and recombinant bIFNs (rbIFNs) were produced by 293 cells. Bovine uterine epithelial or MDBK cells were cultured in the presence or absence of increasing concentrations of each rbIFN for 24, 48, or 72 h. Transcript levels of the IFN-stimulated genes (ISGs) ISG12, ISG15, MX1, and MX2 were analyzed using quantitative reverse transcription-polymerase chain reaction. These messenger RNAs were up-regulated by rbIFN in a time- and concentration-dependent manner. In the epithelial cells, the ISG12 transcript level increased at 48 h after rbIFN treatment but slightly decreased at 72 h, whereas the transcript level of ISG15 increased at 24 h and was maintained through 72 h. Expressions of MX1 and MX2 increased at 72 h after rbIFN treatment. MX1 expression increased in all treatment groups, but MX2 increased only by bIFNTc1. In MDBK cells, the expression of ISG12 was increased by bIFNT1 and bIFNTc1 after 24 and 72 h; however, it was unchanged by rbIFNA. ISG15 increased following the same pattern as that seen in uterine epithelial cells, and MX1 showed a similar expression pattern. MX2 expression was increased by bIFNTc1 treatment in uterine epithelial cells, and its expression was increased by both bIFNT1 and bIFNTc1 in MDBK cells. These results show that epithelial and MDBK cell responses to IFNs differ, suggesting that IFNs possess common functions, but may have acquired different functions following gene duplication.
김민수,민관식,성환후,김찬란,김동교,Kazuhiko Imakawa,김성우 사단법인 한국동물생명공학회 2016 한국동물생명공학회지 Vol.31 No.4
Multiple interferon tau (IFNT) genes exist in bovine. An antiluteolytic substance secreted by the bovine conceptus and primarily responsible for maternal recognition of pregnancy is bovine trophoblast protein 1 (bIFNT1), a new type I interferon tau (IFNT) genes. The objectives of this research were to investigate whether multiple, distinct gene encode bIFNT1 and other type I bIFNT gene in the bovine genome and to examine expression of bIFNT1 and other bIFNTc1 mRNAs during conceptus development. These transcrips could be regulated through caudalrelatedhomeobox- 2 (CDX2) and ETS2 and/or AP1 (JUN) expression, a transcription factor implicated in the control of cell differentiation in the trophectoderm. The presence of mRNAs encoded by bIFNT1 and type I bIFNTc1 genes were examined quantitatively via reverse transcription-polymerase chain reaction (RT-PCR) analysis of total cellular RNA (tcRNA) extracted from on day 17, 20 and 22 bovine conceptuses. The expression level of bIFNT1 was higher on day 17 transcripts were gradually weakly detectable on day 20 and 22. However, the other bIFNTc1 gene examined transcripts was highly expressed on day 20 and transcripts were weakly detectable on day 17 and 22 bovine conceptuses. Furthermore, human choriocarcinoma JEG3 was co-transfected with an -1kb-bIFNT1/c1-Luc constructs and several transcription factor expression plasmids. Compared to each -1kb-bIFNT1/c1-Luc increased when this constructs were co-transfected with, ETS2, AP1(JUN), CREBBP and/or CDX2. Also, bIFNTc1 gene was had very effect on activity by alone ETS2, and AP1 (JUN) expression factors in choriocarcinoma JEG3 cell. However, bIFNT1 gene expression of the upstream region was not identified. We demonstrated that the activities of bIFN genes are regulated by differential, tissue-specific and developmental competence during pregnancy.
Min-Su Kim,Kwan-Sik Min,Hwan Hoo Seong,Kazuhiko Imakawa,Sung Woo Kim 한국수정란이식학회 2016 한국수정란이식학회 학술대회 Vol.2016 No.10
Interferon tau (IFNT), produced by the mononuclear trophectoderm, signals the process of maternal recognition of pregnancy in ruminants, but its expression in vivo is not well characterized. Objectives of this study were to determine IFNT gene isoforms expressed in the bovine uterus, and to identify differences in promoter sequences of IFNT genes that differ in their expression. Through the RNA-seq analysis of bovine conceptuses on days 17, 20 and 22 (day 0 = day of estrus), the expression of only two IFNT transcripts, IFNT1 and IFNTc1, were found, which were indeed classified into the IFNT gene clade. IFNT mRNAs were highest on day 17, and then decreased on days 20, and 22, which were also supported by the results of quantitative RT-PCR. Bovine ear-derived fibroblast (EF) cells were then cotransfected with luciferase reporter constructs carrying 5‘-upstream (positions -1000 to +51) regions of IFNT1 or IFNTc1 and various transcription factor expression plasmids. CDX2, either alone or with other Ap-1, ETS2 and/or CREBBP transcription factors, was found to increase luciferase activity approximately 10 and 18 fold more than twice of those cotransfected with bIFNT1, c1-Luc construct. Furthermore, The degree of transcriptional activation by a combination of the AP1, ETS2, CREBBP and/or CDX2 expression vectors was similar to that of CDX2 along plasmid. However, expression patterns of these Luc activity differented. Whereas bIFNTc1-Luc showed lowest antivity had than bIFNT1-Luc reports. Although, lowest antivity had of the bIFNTc1 –Luc report, cotransfected with the bIFNTc1-Luc construct and AP1(JUN) or/and ETS2 expression plasmid, Luc activity was enhanced approximately 2 and 4-fold more than the bIFNT1-Luc. Furthermore, along CDX2 expression factor had high effect on activity of bIFNT1-Luc reporter than the c1 gene in EF cells. These results suggest that two forms of IFNT genes are expressed in utero and their transcriptional regulations differ.
Min-Su Kim,Kwan-Sik Min,Hwan Hoo Seong,Kazuhiko Imakawa,Sung Woo Kim 한국수정란이식학회 2016 한국수정란이식학회 학술대회 Vol.2016 No.10
Multiple interferon tau (IFNT) genes exist in bovine. An antiluteolytic substance secreted by the bovine conceptus and primarily responsible for maternal recognition of pregnancy is bovine trophoblast protein 1 (bIFNT1), one of new type I interferon tau (IFNT) genes. The objectives of this research were to investigate whether multiple, distinct gene encode bIFNT1 and other type I bIFNT gene in the bovine genome or not and to examine the expression of bIFNT1 and other bIFNTc1 mRNAs during conceptus development. The transcription of these genes could be regulated through caudal-related homeobox-2 (CDX2) and ETS2 and/or AP1(JUN) expression, a transcription factor implicated in the control of cell differentiation in the trophectoderm. The presence of mRNAs encoded by bIFNT1 and type I bIFNTc1 genes were examined quantitatively via reverse transcription-polymerase chain reaction (RT-PCR) analysis of total cellular RNA (tcRNA) extracted from on the days 17, 20 and 22 bovine conceptuses. bIFNT1 was highly expressed on the day 17 and transcripts were gradually and weakly detectable on the days 20 and 22. However, the other bIFNTc1 gene examined transcripts was highly expressed on the day 20 and transcripts were weakly detectable on the days 17 and 22 bovine conceptuses. Furthermore, human choriocarcinoma JEG3 was co-transfected with an -1kb-bIFNT1/c1-Luc constructs and several transcription factor expression plasmids. Compared to each -1kb-bIFNT1/c1-Luc increased when this constructs were co-transfected with, ETS2, AP1(JUN), CREBBP and/or CDX2. Also, bIFNTc1 gene was had higher effect on activity by alone ETS2, and AP1(JUN) expression factors in choriocarcinoma JEG3 cell. However, bIFNT1 gene expression of the upstream region was not idented. These results demonstrate that these genes display differential, tissue-specific expression and developmental regulation during pregnancy.
Identification and characterization of full-length vps29 gene in five mammalian species
Young-Hyun Kim,Sang-Je Park,Jae-Won Huh,Kyoung-Min Kim,이상래,Kazuhiko Imakawa,장규태 한국유전학회 2011 Genes & Genomics Vol.33 No.5
Retromer is a heteropentameric complex associated with retrograde transport of cargo protein from the endosome to the trans-Golgi network. The mammalian retromer complex consists of three vps genes (vps26, vps35, and vps29) and two sorting nexin genes (snx1 and snx2). Previous studies had reported the protein sorting functions of retromer in the intracellular compartment. However, individual genes of retromer complex have not yet been fully characterized. In this study,we identified full-length vps29 gene in human, crab-eating macaque, mouse, rat, and dog species. Total forty-four novel transcripts of vps29, including two actively expressed major transcripts, were identified by 5′- and 3′-RACE. Comparative analysis indicated that functionally important sites of the vps29gene were well conserved in the eukaryotic genome. However,two major transcript variants were occurred in the vertebrate genome only. From our results, we assumed that there are many different transcripts variants of vps29 gene and specifically two major transcripts could play important roles in the protein sorting mechanism.
Human Keratin 14 Oriven HPV 16 E6/E7 Transgenic Mice Exhibit Hyperkeratinosis
Sung-Hyun Kim,Kil-Soo Kim,Eun-Ju Lee,Myoung-Ok Kim,Jun-Hong Park,Kyoung-In Cho,Kazuhliko Imakawa,Byung-Hwa Hyun,Kyu-Tae Chang,Hoon Taek Lee,Zae-Young Ryoo 한국동물생명공학회(구 한국동물번식학회) 2004 Reproductive & Developmental Biology(Supplement) Vol.28 No.2s