Identification of a factor in HeLa cells specific for an upstream transcriptional control sequence of an EIA-inducible adenovirus promoter and its relative abundance in infected and uninfected cells.
Identification of a factor in HeLa cells specific for an upstream transcriptional control sequence of an EIA-inducible adenovirus promoter and its relative abundance in infected and uninfected cells.
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鉴定 HeLa 细胞中 EIA 诱导型腺病毒启动子上游转录控制序列特异的因子及其在感染和未感染细胞中的相对丰度。
DOI：
10.1073/pnas.83.16.5914
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发表时间：
1986
影响因子：
11.1
通讯作者：
Thimmappaya,B
中科院分区：
文献类型：
--
作者：
SivaRaman,L;Subramanian,S;Thimmappaya,B
利用凝胶电泳/DNA结合实验，在HeLa细胞核提取物中检测到eiia诱导腺病毒早期启动子上游转录控制序列特异性因子。通过DNA结合分析和甲基化干扰实验对该启动子的连锁扫描突变体进行分析，发现该因子与位于帽位上游-66和-82之间的17个核苷酸序列5‘ tggagatgacgtagttt3 ’结合。该序列已被证明是该启动子转录所必需的。研究发现，在将EIA蛋白诱导至高水平的条件下，eiia早期启动子特异性因子在未感染的HeLa细胞和感染野生型腺病毒或eiia缺失突变体dl312的细胞中以相当的水平存在[感染后7或20小时，存在阿拉伯糖核苷（阿拉伯糖胞嘧啶）]。基于DNA结合测定的定量，似乎eiia早期启动子的eiia激活转录机制不涉及该因子数量的净变化。
Utilizing the gel electrophoresis/DNA binding assay, a factor specific for the upstream transcriptional control sequence of the EIA-inducible adenovirus EIIA-early promoter has been detected in HeLa cell nuclear extract. Analysis of linker-scanning mutants of the promoter by DNA binding assays and methylation-interference experiments show that the factor binds to the 17-nucleotide sequence 5' TGGAGATGACGTAGTTT 3' located between positions -66 and -82 upstream from the cap site. This sequence has been shown to be essential for transcription of this promoter. The EIIA-early-promoter specific factor was found to be present at comparable levels in uninfected HeLa cells and in cells infected with either wild-type adenovirus or the EIA-deletion mutant dl312 under conditions in which the EIA proteins are induced to high levels [7 or 20 hr after infection in the presence of arabinonucleoside (cytosine arabinoside)]. Based on the quantitation in DNA binding assays, it appears that the mechanism of EIA-activated transcription of the EIIA-early promoter does not involve a net change in the amounts of this factor.