INTERFERON-INDUCED TRANSCRIPTION OF A GENE ENCODING A 15-KDA PROTEIN DEPENDS ON AN UPSTREAM ENHANCER ELEMENT
INTERFERON-INDUCED TRANSCRIPTION OF A GENE ENCODING A 15-KDA PROTEIN DEPENDS ON AN UPSTREAM ENHANCER ELEMENT
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DOI：
10.1073/pnas.84.18.6394
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发表时间：
1987-09-01
影响因子：
11.1
通讯作者：
DARNELL, JE
中科院分区：
文献类型：
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作者：
REICH, N;EVANS, B;DARNELL, JE
已从基因组文库中分离出编码干扰素诱导的15-kDa蛋白的人基因。该基因似乎是单拷贝的，由两个外显子组成，其中第一个外显子含有ATG翻译起始密码子。体外细胞核连续试验表明，干扰素治疗后，该基因的转录速率被刺激。为了分析转录调控序列，我们构建了用于HeLa细胞瞬时转染试验的重组质粒。 发现在转录起始位点的5“处含有115个核苷酸的构建体完全可被干扰素诱导。对缺失突变体的分析鉴定了位于CCAAT盒上游-115和-96之间的干扰素诱导的关键元件。此外，当以基因上游或基因下游的任一方向克隆时，包括该区域的DNA片段可以赋予异源启动子（胸苷激酶）干扰素诱导作用。这些是增强子元件的特性，增强子元件仅在用干扰素治疗后才有活性。这一调控序列可能被一组干扰素诱导的基因所共有，因为在另一个干扰素诱导的基因的RNA起始位点附近的功能区域内存在非常相似的序列。
A human gene encoding an interferon-induced 15-kDa protein has been isolated from a genomic library. The gene appears to be single-copy and is composed of two exons, the first of which contains the ATG translation initiation codon. In vitro nuclear run-on assays showed that the transcription rate of the gene is stimulated after interferon treatment. To analyze transcriptional regulatory sequences, we constructed recombinant plasmids for use in transient transfection assays of HeLa cells. Constructs containing 115 nucleotides 5'' to the transcription initiation site were found to be fully inducible by interferon. Assays of deletion mutant identified a critical element for interferon induction located between -115 and -96, just upstream of the "CCAAT box." Moreover, a DNA fragment including this region can confer interferon inducibility on a heterologous promoter (thymidine kinase) when cloned in either orientation upstream of the gene or drownstream of the gene. These are properties characteristic of an enhancer element that is active only after treatment with interferon. This regulatory sequence may be shared by a group of interferon-induced genes, since a very similar sequence is present within the functional region near the RNA start site of another interferon-induced gene.