Faculty Profiles - EGUCHI Takanori

Papers - EGUCHI Takanori

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遺伝子発現の抑圧的調節を仲介するマウスctgf3'-UTR segmentの性質

近藤誠二, 久保田聡, 江口傑徳, 服部高子, 中西徹, 菅原利夫, 滝川正春

日本口腔科学会雑誌 50 ( 6 ) 568 - 568 2001.11

Cell-type-specific trans-activation of herpes simplex virus thymidine kinase promoter by the human T-cell leukemia virus type I Tax protein. Reviewed

Kubota S, Mukudai Y, Hattori T, Eguchi T, Kondo S, Takigawa M

DNA and cell biology 20 ( 9 ) 563 - 568 2001.9

Publishing type：Research paper (scientific journal)

The human T-cell leukemia virus type I Tax protein (HTLV-I Tax) is known as a trans-activating factor for a variety of genes, including those of cytokines. Here, we show that Tax is capable of activating the herpes simplex virus thymidine kinase (HSV-TK) promoter in certain mammalian cell lines. In murine NIH 3T3 fibroblasts and human HeLa cells, trans-activation by Tax was remarkably strong, whereas in human chondrocytic HCS-2/8 and monkey kidney Cos-7 cells, the responsiveness of the TK promoter to Tax was poor. Deletion analysis revealed that one of the two previously described Sp1 sites is required for the Tax responsiveness, whereas the CTF binding site is not. The results suggest possible interactions between the oncogenic Tax protein and the viral TK in coinfected cells in vivo. Care should be taken in the context of HTLV-I research, as the HSV-TK promoter has been widely used in molecular biology and gene therapeutics.

DOI： 10.1089/104454901317094972

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多機能成長因子CTGF/Hcs24遺伝子の転写後制御エレメント,CAESARの作用機序

久保田聡, 近藤誠二, 椋代義樹, 江口傑徳, 服部高子, 中西徹, 滝川正春

生化学 73 ( 8 ) 710 - 710 2001.8

Regulatory Mechanism of Human Connective Tissue Growth Factor (CTGF/Hcs24) Gene Expression in a Human Chondrocytic Cell Line, HCS-2/8 Reviewed

T. Eguchi, S. Kubota, S. Kondo, T. Shimo, T. Hattori, T. Nakanishi, T. Kuboki, H. Yatani, M. Takigawa

Journal of Biochemistry 130 ( 1 ) 79 - 87 2001.7

Novel mode of processing and secretion of connective tissue growth factor/ecogenin（CTGF/Hcs24） in chondrocytic HCS-2/8 cells Reviewed

S. Kubota, T. Eguchi, T. Shimo, T. Nishida, T. Hattori, S. Kondo, T. Nakanishi, M. Takigawa

Bone 29 ( 2 ) 155 - 161 2001

Publishing type：Research paper (scientific journal)

The synthesis, processing, and secretion of human connective tissue growth factor (CTGF/Hcs24) in a human chondrocytic cell line, HCS-2/8, were analyzed immunochemically. By metabolic pulse-labeling, chasing, and subsequent immunoprecipitation analyses, active synthesis of CTGF was observed not only in growing HCS-2/8 cells, but also in confluent cells. However, secretion and processing of CTGF were found to be regulated differentially, depending upon the growth status. During phases of growth, HCS-2/8 cells released CTGF molecules immediately without sequestering them within the cell layer. In contrast, after the cells reached confluence, the secretion slowed, resulting in an accumulation of CTGF in the cells or extracellular matrices (ECMs). Also, in confluent cell layers, a 10 kDa protein that was reactive to an anti-CTGF serum was observed. This CTGF-related small protein was not detected immediately after labeling, but gradually appeared within 6 h after chase, which suggests its entity as a processed subfragment of CTGF. Surprisingly, the 10 kDa protein was stable even 48 h after synthesis, and was not released by ECM digestion, suggesting an intracellular maintenance and function. Taken together, the behavior of CTGF in HCS-2/8 cells is remarkably different from that reported in fibroblasts, which may represent unique roles for CTGF in the growth and differentiation of chondrocytes. Copyright © 2001 Elsevier Science Inc.

DOI： 10.1016/S8756-3282(01)00492-6

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Characterization of a mouse ctgf 3 '-UTR segment that mediates repressive regulation of gene expression

S Kondo, S Kubota, T Eguchi, T Hattori, T Nakanishi, T Sugahara, M Takigawa

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 278 ( 1 ) 119 - 124 2000.11

Language：English Publisher：ACADEMIC PRESS INC

We isolated a small segment of the 3'-untranslated region (3'-UTR) in the mouse connective tissue growth factor (ctgf/fisp12) gene and evaluated its functionality. Comparison of the nucleotide sequences of human and mouse ctgf 3'-UTRs revealed a conserved small segment of 91 bases, The corresponding segments of the 3'-UTRs shared as much as 82.4% homology, whereas the overall homology between the 3'-UTRS was 71.8%. To study the functionality of the conserved segment, the corresponding region of mouse ctgf cDNA was amplified from NIH3T3 cells. When it was fused downstream of a marker gene, it showed remarkable repressive effects on gene expression. The repressive effect of the sense form was more prominent than that of the antisense form. Computer analyses of these sequence predicted stable secondary structures, suggesting that they act at the RNA level. The predicted structures of the sense and antisense forms appeared to be slightly different, which is consistent with the difference in repressive function. These findings defined the conserved small element in the mouse ctgf gene as a potent negative regulator of gene expression, which may act at a posttranscriptional level. (C) 2000 Academic Press.

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Characterization of a Mouse ctgf 3′-UTR Segment That Mediates Repressive Regulation of Gene Expression Reviewed

Seiji Kondo, Satoshi Kubota, Takanori Eguchi, Takako Hattori, Tohru Nakanishi, Toshio Sugahara, Masaharu Takigawa

Biochemical and Biophysical Research Communications 278 ( 1 ) 119 - 124 2000.11

Identification of an RNA element that confers post-transcriptional repression of connective tissue growth factor/hypertrophic chondrocyte specific 24 (ctgf/hcs24) gene: Similarities to retroviral RNA-protein interactions Reviewed

S Kubota, S Kondo, T Eguchi, T Hattori, T Nakanishi, RJ Pomerantz, M Takigawa

ONCOGENE 19 ( 41 ) 4773 - 4786 2000.9

Language：English Publishing type：Research paper (scientific journal) Publisher：NATURE PUBLISHING GROUP

The repressive effect of the 3'-untranslated region (3'-UTR) in human connective tissue growth factor/hypertrophic chondrocyte specific 24 (ctgf/hcs24) mRNA on gene expression had been demonstrated in our previous study. Here, we identified a minimal RNA element in the 3'-UTR, which acts as a cis-acting element of structure-anchored repression (CAESAR). Deletion analyses of the 3'-UTR led us to minimize the element of 84 bases at the junction of the coding region and the 3'-UTR. The minimized RNA segment is predicted, and actually capable of forming a stable secondary structure in vitro. Mutational analyses disclosed a significant relationship between the predicted structure and repressive effect. The utility of CAESAR as a post-transcriptional regulatory element was represented by the fact that steady-state mRNA levels were not affected by CAESAR linked in cis, while protein levels from such a chimeric gene were markedly reduced. Of note, the CAESAR sequence exerted no effect, when it was placed upstream of the promoter. Finally, RNA gel electromobility-shift analyses demonstrated a nuclear factor that interacts with the folded CAESAR. Taken together, it was uncovered that CAESAR of ctgf is a novel post-transcriptional structured RNA regulatory element, probably acting through direct interactions with a nuclear factor as observed in retroviral RNA elements with certain proteins.

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軟骨由来の成長因子CTGF/Hcs24遺伝子の転写後制御エレメント,CAESARの構造機能連関

久保田聡, 近藤誠二, 江口傑徳, 服部高子, 中西徹, 滝川正春

生化学 72 ( 8 ) 972 - 972 2000.8

軟骨由来の成長因子CTGF/Hcs24遺伝子に同定された新たな転写後制御エレメント,CAESAR

久保田聡, 近藤誠二, 江口傑徳, 服部高子, 中西徹, 滝川正春

日本骨代謝学会雑誌 18 ( 2 ) 119 - 119 2000.6

軟骨細胞および線維芽細胞株におけるCTGF/Ecogenin遺伝子発現の3'-UTRによる調節

近藤誠二, 久保田聡, 江口傑徳, 服部高子, 中西徹, 菅原利夫, 滝川正春

岡山歯学会雑誌 19 ( 1 ) 205 - 206 2000.6

アデノウイルスを用いた顎関節への遺伝子導入の試み

縄稚久美子, 窪木拓男, 完山学, 藤沢拓生, 園山亘, 江口傑徳, 上原淳二, 矢谷博文, 山下敦

日本補綴歯科学会雑誌 43 ( 102回特別 ) 180 - 180 1999.10

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