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      • SCOPUSKCI등재

        TPA로 분화된 U937 세포에서 사람 세포거대바이러스에 의한 c-jun Promoter 활성도의 변화

        박정규,김대중,김진희,한태희,황응수,최명식,국윤호,최성배,차창룡,Park, Chung-Gyu,Kim, Dae-Joong,Kim, Jin-Hee,Han, Tae-Hee,Hwan, Eung-Soo,Choi, Myong-Sik,Kook, Yoon-Hoh,Choi, Sung-Bae,Cha, Chang-Yong 대한미생물학회 1999 Journal of Bacteriology and Virology Vol.28 No.1

        Transient transfection assay has been done to evaluate whether the c-jun activation would be prerequisite to the induction of permissiveness against human cytomegalovirus using in vitro cell model in which U937 has been induced to express CD11b and CD14 to become potential monocyte/macrophage cells by TPA treatment. U937 cells were treated with $10\;{\mu}M$, $50\;{\mu}M$ or $100\;{\mu}M$ of TPA. The cell morphology change was observed and the expression of the CD11b and CD14 was confirmed by FACS. Differentiated cells were transfected with pJLuc reporter vector which contained the wild type murine c-jun promoter spanning the SP1, CTF, ATF/CREB and MEF-2 binding sites upstream of the firefly luciferase gene. After 48 hrs of transfection, the cells were infected with HCMV Towne strain and the luciferase activity was assessed at 1 hand 4 h pi. The transfection assay showed no activation of the c-jun promoter at 1 h pi, instead, it showed 2 times increase of the its activity at 4 h pi. There was no difference of the c-jun promoter activation between TPA treated and untreated U937 cells, implying that c-jun activation might not be prerequisite for allowing cells to be premissive to HCMV, although HCMV infection itself could activate c-jun promoter.

      • SCOPUSKCI등재

        사람세포거대바이러스 (Human CVytomegalovirus)의 극초기항원-1 (Immediate Early-1, IE-1)에 방응하는 c-jun Promoter의 유전자 지도 분석

        박정규(Chung Gyu Park),한태희(Tae Hee Han),김대중(Dae Joong Kim),김진희(Jin Hee Kim),황응수(Eung Soo Hwang),최성배(Sung Bae Choi),차창룡(Chang Yong Cha) 대한바이러스학회 1998 Journal of Bacteriology and Virology Vol.28 No.3

        Human cytomegalovirus (HCMV) has the ability to activate the expression of many viral and cellular genes. Among various viral proteins, the immediate early proteins (IE1-7ZkDa, IE2- 86kDa) have been known to be potent transactivators. The product of c-jun proto-oncogene is important in cell activation and differentiation. Here, we tried to find out if the IE could activate the c-jun promoter and also tried to identify the responsible sequence elements in the c-jun activation by IE1-72kDa. We found HCMV IE expression transactivated the c-jun promoter in human embryonal lung fibroblasts (HEL). The activation fold by JE1-72kDa, IE2-86kDa and IE2-55kDa was 23, 35, and 5, respectively. When the expression of each IE was combined, it showed synergism. Expression of (IE1-72kDa + IE2-86kDa) and (IE1-72kDa + IE2-86kDa + IE2-55kDa) resulted in 131 and 162 fold increase, respectively. The c-jun promoter region between -117 and -59 contains binding sites for the transcription factors Spl, CAAT, AP-1 like (ATF/CREB), and MEF2. Transient expression assays were performed using various reporter plasmids containing the c-jun promoter-regulatory region linked to the luciferase gene and a plasmid expressing HCMV IE1 gene. Deletional and point mutational analysis showed that the sequence between -225 to -160 and the CTF binding site were involved in the up-regulation of c-jun promoter.

      • SCOPUSKCI등재

        TPA로 분화된 U937 세포에서 사람 세포거대바이러스에 의한 c- jun Promoter 활성도의 변화

        박정규(Jung Gyu Park),김대중(Dae Joong Kim),김진희(Jin Hee Kim),한태희(Tae Hee Han),황응수(Eung Soo Hwang),최명식(Myung Sik Choi),국윤호(Yoon Ho Gook),최성배(Sung Bae Choi),차창룡(Chang Ryong Cha) 대한바이러스학회 1999 Journal of Bacteriology and Virology Vol.29 No.2

        Transient transfection assay has been done to evaluate whether the c-jun activation would be prerequisite to the induction of permissiveness against human cytomegalovirus using in vitro cell model in which U937 has been induced to express CD11b and CD14 to become potential monocyte/macrophage cells by TPA treatment. U937 cells were treated with 10 ㎛, 50 ㎛ or 100 ㎛ of TPA. The cell morphology change was observed and the expression of the CD11b and CD14 was confirmed by FACS. Differentiated cells were transfected with pJLuc reporter vector which contained the wild type murine c-jun promoter spanning the SP1, CTF, ATF/CREB and MEF-2 binding sites upstream of the firefly luciferase gene. After 48 hrs of transfection, the cells were infected with HCMV Towne strain and the luciferase activity was assessed at 1 h and 4 h pi. The transfection assay showed no activation of the c-jun promoter at 1 h pi, instead, it showed 2 times increase of the its activity at 4 h pi. There was no difference of the c-jun promoter activation between TPA treated and untreated U937 cells, implying that c-jun activation might not be prerequisite for allowing cells to be premissive to HCMV, although HCMV infection itself could activate c-jun promoter.

      • KCI등재

        Suppression of Thrombospondin-1 Expression by PMA in the Porcine Aortic Endothelial Cells

        Chang, Seo-Yoon,Kang, Jung-Hoon,Hong, Kyong-Ja Korean Society of Life Science 2004 생명과학회지 Vol.14 No.1

        암의 성장과 신생혈관 억제인자로 알려진 thrombospondin-1의 생합성은 다양한 외부자극에 대해 전사단계에서 세포 특이적으로 조절된다. 이전의 연구에서 본 연구자들은 PMA가 정상 돼지 대동맥 내피세포(PAE)에서는 TSP-1의 발현을 감소시키는 반면 사람 간암 세포주인 Hep3B에서는 증가시키는 사실을 발견하였다. PMA 처치에 따른 정상 돼지 대동맥 내피세포에서의 TSP-1의 발현 감소현상은 tsp-1 유전자 조절부위의 염기서열 -767과 -723사이에 존재하는 염기서열이 억제 부위임을 밝혀 이러한 결과를 바탕으로 -767에서 -723 염기서열을 서로 부분 중복되도록 세 종류의 올리고 탐식자 (올리고 탐식자 a-1, -767∼-738; 올리고 탐식자 a-2, -759∼-730; 올리고 탐식자 a-3, -752∼723)를 제작하여 -767과 -723 부위의 특정 염기서열과 이에 결합하는 인자를 EMSA을 수행하여 분석하였다. 실험 결과, PMA 처치에 따른 정상 돼지 대동맥 내피세포에서의 TSP-1 감소는 -752에서 -730 사이의 염기서열이 저해 조절인자와 결합함과 더불어 -767에서 -760과 -752에서 -730 사이의 염기서열들에 촉진 조절인자들이 결합하지 못함으로서 기인된다는 실험적 사실을 관찰하였고. 특히, PMA 처치는 정상 돼지 대동맥 내피세포에서 저해 조절인자의 -752에서 -730 부위에 대한 친화력을 향상시켰으며 이러한 친화력은 c-Jun 항체에 의해 영향을 받지 않았다. Thrombospondin-1 (TSP-1), a negative regulator in tumor growth and angiogenesis, is cell-type specifically regulated and at transcriptional level by external stimuli. Previously, we found that phorbol 12-myristate 13-acetate (PMA) suppressed TSP-1 expression in porcine aortic endothelial (PAE) cell, but enhanced in hepatoma cell line, Hep 3B cell. A region between -767 and -723 on the tsp-1 promoter was defined as a responsive site to the suppression in PAE cell. eased on the previous results, the molecular mechanism of TSP-1 expression was determined by characterizing interactions between cis-elements and trans-factors using three overlapped oligonucleotide probes, oligo a-1 (from -767 to -738), a-2 (-759 to -730) and a-3 (-752 to -723). The results from electromobility shift assay showed that PMA-induced suppression of TSP-1 transcription in PAE cell might be caused via a negative regulator binding to the region from -752 to -730 and additionally generated by lacking two positive regulators binding to the sites from -767 to -760 and from -752 to -730. Especially, PMA enhanced the binding ability of the negative regulator to the site from -752 to -730 in PAE cell, but anti-c-Jun did not affected its binding ability.

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