Real-time study of interactions between a composite DNA regulatory region (HIV-1 LTR NRE) and several transcription factors of nuclear extracts.
Real-time study of interactions between a composite DNA regulatory region (HIV-1 LTR NRE) and several transcription factors of nuclear extracts.
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实时研究复合 DNA 调控区 (HIV-1 LTR NRE) 与核提取物的几种转录因子之间的相互作用。
DOI：
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发表时间：
1999
期刊：
影响因子：
--
通讯作者：
C. Vaquero
中科院分区：
文献类型：
--
作者：
L. Galio;S. Briquet;C. Vaquero
在这里，我们描述了第一次实时研究核蛋白与复合DNA调节区的相互作用。我们研究了HIV-1 LTR的负调控元件(NRE)的三个靶点之间的相互作用，包括重叠两个负调节区USF和NFIL-6的非规范GATA位点，以及它们在核提取液中对应的转录因子。通过带移分析，LTRNRE和不同的核提取物之间没有检测到GATA结合活性，尽管体外足迹证明了这一点。此外，LTRNRE和一个USF寡核苷酸显示出相同的延迟复合体。对这些相互作用的Biaccore研究表明，huGATA-3以及USF与固定的LTRNRE寡核苷酸结合。用GATA、USF和NFIL-6寡核苷酸进行的竞争分析清楚地表明，该调节区可以结合huGATA-3和USF因子。最后，用特异性抗huGATA-3和抗USF2多克隆抗体确定与LTRNRE形成的复合体中是否存在USF和huGATA-3蛋白。
Here we describe the first real-time study of nuclear protein interaction with a composite DNA regulatory region. We studied the interplay between the three target sites of the negative regulatory element (NRE) of HIV-1 LTR, comprising a noncanonical GATA site overlapping two negative regulatory regions, USF and NFIL-6, and their corresponding transcription factors in nuclear extracts. By bandshift analysis, no GATA binding activity could be detected between LTR NRE and different nuclear extracts, although evidenced by in vitro footprinting. Additionally, the LTR NRE and a USF oligonucleotide showed identical retarded complexes. BIAcore study of these interactions revealed the binding of huGATA-3, as well as USF, to the immobilized LTR NRE oligonucleotide. Competition analyses, performed with GATA, USF, and NFIL-6 oligonucleotides, clearly showed that this regulatory region could bind both huGATA-3 and USF factors. Finally, the presence of USF and huGATA-3 proteins in the complexes formed with LTR NRE was ascertained using specific anti-huGATA-3 and anti-USF2 polyclonal antibodies.
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影响因子：
10.5
作者：
S. C. Williams;Carrie A. Cantwell;P. Johnson
通讯作者：
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DOI：
10.1016/s0021-9258(18)37883-9
发表时间：
1988-08
期刊：
The Journal of biological chemistry
影响因子：
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作者：
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影响因子：
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作者：
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通讯作者：
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