DNA residence time is a regulatory factor of transcription repression.
DNA residence time is a regulatory factor of transcription repression.
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DOI：
10.1093/nar/gkx728
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发表时间：
2017-11-02
影响因子：
14.9
通讯作者：
Gebhardt JCM
中科院分区：
文献类型：
--
作者：
Clauß K;Popp AP;Schulze L;Hettich J;Reisser M;Escoter Torres L;Uhlenhaut NH;Gebhardt JCM
转录包括一系列高度调控的内在随机过程，导致转录的爆发被静止所打断。在转录激活或抑制中，转录因子动态地与DNA结合，每个因子具有独特的停留时间。DNA停留时间在转录过程中是否重要尚不清楚。在这里，我们设计了一系列的转录抑制剂不同的DNA停留时间，利用模块化的DNA结合结构域的转录激活因子样效应（TALE）和不同的核苷酸识别重复结构域的数量。我们使用单分子追踪来表征我们的阻遏物在活细胞中的DNA停留时间。的停留时间依赖于非线性的重复域的数量和不同的超过6倍。这些因素引起糖皮质激素受体激活基因SGK 1的转录水平的停留时间依赖性降低。下调转录是由于较低的突发频率在长结合阻遏物的存在下，是根据内源性激活剂结合的竞争性抑制模型。我们的单分子实验揭示了转录因子DNA停留时间作为控制转录抑制的调控因子，并建立了TALE-DNA结合结构域作为转录调控的时间解剖的工具。
Transcription comprises a highly regulated sequence of intrinsically stochastic processes, resulting in bursts of transcription intermitted by quiescence. In transcription activation or repression, a transcription factor binds dynamically to DNA, with a residence time unique to each factor. Whether the DNA residence time is important in the transcription process is unclear. Here, we designed a series of transcription repressors differing in their DNA residence time by utilizing the modular DNA binding domain of transcription activator-like effectors (TALEs) and varying the number of nucleotide-recognizing repeat domains. We characterized the DNA residence times of our repressors in living cells using single molecule tracking. The residence times depended non-linearly on the number of repeat domains and differed by more than a factor of six. The factors provoked a residence time-dependent decrease in transcript level of the glucocorticoid receptor-activated gene SGK1. Down regulation of transcription was due to a lower burst frequency in the presence of long binding repressors and is in accordance with a model of competitive inhibition of endogenous activator binding. Our single molecule experiments reveal transcription factor DNA residence time as a regulatory factor controlling transcription repression and establish TALE-DNA binding domains as tools for the temporal dissection of transcription regulation.
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