Identification of differentially expressed nucleolar TGF-beta1 target (DENTT) in human lung cancer cells that is a new member of the TSPY/SET/NAP-1 superfamily.
Identification of differentially expressed nucleolar TGF-beta1 target (DENTT) in human lung cancer cells that is a new member of the TSPY/SET/NAP-1 superfamily.
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鉴定人肺癌细胞中差异表达的核仁 TGF-β1 靶标 (DENTT)，该细胞是 TSPY/SET/NAP-1 超家族的新成员。
作者：
L. Ozbun;Liang You;Sharon Kiang;J. Angdisen;Alfredo Martínez;S. Jakowlew
以转化生长因子-β1(TGF-β1)反应性上皮性非小细胞肺癌(NSCLC)细胞系NCI-H727为研究对象，筛选与转化生长因子-β1(TGF-β1)相关的潜在靶基因。通过差异显示，比较了转化生长因子-β1处理的细胞和赋形剂处理的细胞之间的差异表达模式。一个496个碱基的片段被转化生长因子-β1差异地增加了三倍，并且通过Northern分析与NCI-H727细胞中2.7kb的mRNA杂交，显示与任何已知的基因序列都没有显著的匹配。这个新基因的转录产物被我们命名为差异表达的核仁转化生长因子-β1靶基因(DENTT)，它在几个正常人体组织中都有表达，其中在脑中的表达水平最高。人脑cDNA文库筛选和5‘端快速扩增获得全长DENTT基因，含有一个1899个碱基的开放阅读框，编码633个氨基酸的蛋白质，具有四个潜在的核定位信号(NLSS)和两个卷曲的区域。DENTT包含一个保守的191个残基结构域，它与TSPY/TSPY-like/Set/NAP-1超家族有重要的同一性和定义。增强型绿色荧光蛋白(EGFP)标记的全长DENTT转染COS-7细胞后显示核仁和胞浆定位。将EGFP标记的DENTT NLS缺失的缺失构建体从核仁中排除。虽然NLS-1对于DENTT的核仁定位是必要的，但对于单一的核仁定位是不够的。我们的数据表明，在对转化生长因子-β1有反应的非小细胞肺癌细胞中，转化生长因子-β1诱导DENTT基因的表达与其诱导转化生长因子-β1的表达、诱导细胞外基质基因的表达以及抑制软琼脂糖集落形成有关。在无反应的非小细胞肺癌细胞中，转化生长因子-β1不诱导DENTT基因的表达。我们的数据表明，这个新的转化生长因子-β1靶基因具有不同的结构域，可以定向到不同的亚核位置。
The transforming growth factor-beta1 (TGF-beta1) responsive epithelial non-small-cell lung cancer (NSCLC) cell line NCI-H727 was used to identify potential target genes involved in TGF-beta1-mediated responses. Comparative cDNA expression patterns between cells treated with TGF-beta1 and those treated with vehicle were generated by differential mRNA display. One 496-bp fragment, differentially increased threefold by TGF-beta1 and hybridizing to a 2.7-kb mRNA species in NCI-H727 cells by Northern analysis, revealed no significant match to any known gene sequence. The mRNA transcript of this novel gene that we named differentially expressed nucleolar TGF-beta1 target (DENTT) is expressed in several normal human tissues, with the highest level of expression in brain. Human brain cDNA library screening and 5' rapid amplification of cDNA ends yielded full-length DENTT cDNA containing an 1899-bp open reading frame encoding a predicted 633-amino-acid protein with four potential nuclear localization signals (NLSs) and two coiled-coil regions. DENTT contains a conserved 191-residue domain that shows significant identity to, and defines, the TSPY/TSPY-like/SET/NAP-1 superfamily. Enhanced green fluorescent protein (EGFP)-tagged full-length DENTT transfected into COS-7 cells showed nucleolar and cytoplasmic localization. Transfection of EGFP-tagged DENTT NLS deletion constructs lacking the bipartite NLS-1 were excluded from the nucleolus. While NLS-1 is necessary for nucleolar localization of DENTT, it is not sufficient for sole nucleolar localization. Our data show that DENTT mRNA induction by TGF-beta1 correlates with induction of TGF-beta1 mRNA, induction of extracellular matrix gene expression, and inhibition of colony formation in soft agarose in TGF-beta1 responsive NSCLC cells when exposed to TGF-beta1. TGF-beta1 does not induce DENTT mRNA expression in TGF-beta1 nonresponsive NSCLC cells. Our data suggest that this novel TGF-beta1 target gene has distinct domains for direction to different subnuclear locations.
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影响因子：
--
作者：
G. Pfeifer;A. Riggs
通讯作者：
G. Pfeifer;A. Riggs
影响因子：
2
作者：
J. Prins;E. de Vries;N. Mulder
通讯作者：
J. Prins;E. de Vries;N. Mulder
DOI：
10.1016/s0021-9258(18)54461-6
发表时间：
1991-12
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
M. Keeton;S. Curriden;A. J. Zonneveld;D. Loskutoff
通讯作者：
M. Keeton;S. Curriden;A. J. Zonneveld;D. Loskutoff
DOI：
10.1016/s0021-9258(18)69042-8
发表时间：
1988-03
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
J. Keski‐oja;R. Raghow;M. Sawdey;D. Loskutoff;A. Postlethwaite;A. Kang;H. Moses
通讯作者：
J. Keski‐oja;R. Raghow;M. Sawdey;D. Loskutoff;A. Postlethwaite;A. Kang;H. Moses
影响因子：
2.9
作者：
CHAN, WY;LIU, QR;CHAN, PK
通讯作者：
CHAN, PK