The interferon-stimulated gene 54 K promoter contains two adjacent functional interferon-stimulated response elements of different strength, which act synergistically for maximal interferon-alpha inducibility.
The interferon-stimulated gene 54 K promoter contains two adjacent functional interferon-stimulated response elements of different strength, which act synergistically for maximal interferon-alpha inducibility.
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干扰素刺激基因54 K启动子包含两个相邻的不同强度的功能性干扰素刺激反应元件，它们协同作用以获得最大干扰素-α诱导能力。
DOI：
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发表时间：
1994
期刊：
影响因子：
--
通讯作者：
J. Trapman
中科院分区：
文献类型：
--
作者：
H. Bluyssen;R. Vlietstra;A. C. van der Made;J. Trapman
分离了干扰素-α (IFN-α) 调节的仓鼠 ISG-54 K 基因，该基因在仓鼠 CHO-12 细胞中被激活至少 40 倍，并对启动子区域进行了详细表征。序列分析揭示了两个元件的存在，与干扰素刺激反应元件（ISRE）共有序列[AGTTTCNNTTTC(CT)]密切相关。推定的ISRE-I序列(GGTTTCAATTTCT)位于位置-97至-85； ISRE-II (AGTTTTACTTTCT) 与 ISRE-I 在三个位置上不同，被发现直接位于 ISRE-I 上游的 -110 至 -98 位。在 CHO-12 细胞中的瞬时转染测定中，野生型仓鼠 ISG-54K-启动子-氯霉素-乙酰基转移酶 (CAT) 报告构建体显示出 40-80 倍的诱导作用，为研究两种 ISRE 的功能特性提供了一个极好的模型。为了查明这两个元件是否在启动子的干扰素调节中起作用，在-110至-85区域和侧翼序列中引入了选定的点突变。 （突变的）ISG-54 K 启动子与 CAT 报告基因连接，并在鼠 (Mu)IFN-α 6 不存在和存在的情况下在 CHO 细胞中瞬时表达。转染显示，ISG-54 K 启动子的最佳干扰素诱导需要 -97 至 -85 (ISRE-I) 和 -110 至 -98 (ISRE-II) 片段。然而，ISRE-I 的活性比 ISRE-II 强大约七倍。 ISRE-II中不同的三个ISRE-I碱基的顺序取代表明-105位的T导致ISRE-II的活性降低。 ISG-54 K 启动子构建体的转染，其中 ISRE-I 被 ISRE-II 取代，生成具有两个 ISRE-II 片段的启动子，反之亦然（两个 ISRE-I），为这两个元件在 IFN-α 诱导中的作用提供了进一步的证据。重要的是，转染研究中获得的所有数据都表明两种 ISRE 协同合作。协同作用的机制很可能是与 ISRE 结合的转录因子之间的间接相互作用，因为两个 ISRE 的空间排列增加一个完整的螺旋转或半个螺旋并不会导致启动子活性的实质性降低。
The interferon-alpha(IFN-alpha)-regulated hamster ISG-54 K gene, which is activated in hamster CHO-12 cells at least 40-fold, was isolated and the promoter region was characterized in detail. Sequence analysis revealed the presence of two elements, closely related to the interferon-stimulated-response-element (ISRE) consensus sequence [AGTTTCNNTTTC(CT)]. The putative ISRE-I sequence (GGTTTCAATTTCT) is located at position -97 to -85; ISRE-II (AGTTTTACTTTCT), which differs at three positions from ISRE-I, is found directly upstream of ISRE-I at position -110 to -98. In a transient transfection assay in CHO-12 cells the wild-type hamster ISG-54K-promoter-chloramphenicol-acetyl-transferase (CAT) reporter construct showed a 40-80-fold induction, offering an excellent model to study the functional properties of the two ISRE. To find out whether both elements were functional in interferon regulation of the promoter, selected point mutations were introduced in the -110 to -85 region and in flanking sequences. The (mutated) ISG-54 K promoter was linked to the CAT reporter gene and transiently expressed in CHO cells in the absence and presence of murine (Mu)IFN-alpha 6. Transfections showed that both the -97 to -85 (ISRE-I) and the -110 to -98 (ISRE-II) segment were needed for optimal interferon induction of the ISG-54 K promoter. However, ISRE-I has an approximately sevenfold stronger activity compared to ISRE-II. Sequential substitution of the three ISRE-I bases, which differ in ISRE-II showed that the T at position -105 causes the lower activity of ISRE-II. Transfection of ISG-54 K promoter constructs, in which ISRE-I was replaced by ISRE-II, which generates a promoter with two ISRE-II segments, and vice versa (two ISRE-I), provided further evidence for a role of both elements in IFN-alpha induction. Importantly, all data obtained in transfection studies show that the two ISRE cooperate synergistically. The mechanism of synergism is most probably an indirect interaction between transcription factors binding to the ISRE, because an increase in the spacial arrangement of the two ISRE with a complete helical turn or half a turn did not result in a substantial decrease in promoter activity.
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与鼠 (2-5) 寡腺苷酸合成酶 ME-12 基因的干扰素-α/β 反应元件 B 结合的其他因子的分子克隆和表征。
DOI：
10.1073/pnas.89.19.8859
发表时间：
1992
影响因子：
11.1
作者：
Yan,C;Tamm,I
通讯作者：
Tamm,I
DOI：
10.1073/pnas.83.23.8929
发表时间：
1986-12-01
影响因子：
11.1
作者：
LEVY, D;LARNER, A;DARNELL, JE
通讯作者：
DARNELL, JE
影响因子：
10.5
作者：
KESSLER, DS;VEALS, SA;LEVY, DE
通讯作者：
LEVY, DE
DOI：
10.1073/pnas.84.18.6394
发表时间：
1987-09-01
影响因子：
11.1
作者：
REICH, N;EVANS, B;DARNELL, JE
通讯作者：
DARNELL, JE
影响因子：
56.9
作者：
SCHINDLER, C;SHUAI, K;DARNELL, JE
通讯作者：
DARNELL, JE