Multiple protein-DNA interactions in the TATAA-less mouse thymidylate synthase promoter.
Multiple protein-DNA interactions in the TATAA-less mouse thymidylate synthase promoter.
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无 TATAA 的小鼠胸苷酸合酶启动子中的多种蛋白质-DNA 相互作用。
DOI：
10.1093/nar/19.9.2267
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发表时间：
1991
影响因子：
14.9
通讯作者：
Johnson,LF
中科院分区：
文献类型：
--
作者：
Jolliff,K;Li,Y;Johnson,LF
小鼠胸苷酸合酶 (TS) 启动子缺乏 TATAA 盒，可在 60 个核苷酸区域的多个位点指导转录起始。所有对转录重要的序列都位于第一个起始位点附近。对 TS 启动子该区域的各种序列进行凝胶迁移率变化和足迹分析，确定了三种主要的蛋白质-DNA 相互作用。其中一个对应于 Sp1 与第一个转录起始位点下游的非共有结合位点相互作用。通过定点诱变使结合位点失活导致基因表达量减少 3 倍以及转录起始位点分布的显着变化。负责其他两种复合物（CII 和 CII）的蛋白质似乎与之前研究过的任何常见转录因子都不对应。 CII 蛋白与第一个起始位点非常接近地结合。通过定点诱变使结合位点失活对表达影响很小。 CHI 蛋白紧邻 CII 上游结合。该位点失活导致表达量减少 12 倍，表明它对于 TS 基因的表达很重要。
The mouse thymidylate synthase (TS) promoter lacks a TATAA box and directs transcriptional initiation at multiple sites over a 60 nucleotlde region. All of the sequences that are Important for transcription are located within close proximity to the first initiation site. Gel mobility shift and footprlnting analyses with various sequences from this region of the TS promoter identified three major protein-DNA interactions. One of these corresponds to Sp1 interacting with a nonconsensus binding site downstream of the first transcriptional initiation site. Inactivation of the binding site by site-directed mutagenesis led to a 3-fold reduction in gene expression as well as a significant change in distribution of transcriptional start sites. The proteins responsible for the other two complexes (CII and CII) do not appear to correspond to any of the common transcription factors that have been studied previously. The CII protein binds very close to the first initiation site. Inactivation of the binding site by sitedirected mutagenesis had little effect on expression. The CHI protein binds Immediately upstream of CII. Inactivation of this site led to a 12-fold reduction in expression, indicating that it is Important for expression of the TS gene.