Satory Dominik, Gordon Alasdair J E, Wang Mengyu, Halliday Jennifer A, Golding Ido, Herman Christophe
Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX 77030, USA.
Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, TX 77030, USA Graduate Program in Structural and Computational Biology and Molecular Biophysics, Baylor College of Medicine, Houston, TX 77030, USA.
Nucleic Acids Res. 2015 Dec 2;43(21):10190-9. doi: 10.1093/nar/gkv839. Epub 2015 Aug 24.
DksA is an auxiliary transcription factor that interacts with RNA polymerase and influences gene expression. Depending on the promoter, DksA can be a positive or negative regulator of transcription initiation. Moreover, DksA has a substantial effect on transcription elongation where it prevents the collision of transcription and replication machineries, plays a key role in maintaining transcription elongation when translation and transcription are uncoupled and has been shown to be involved in transcription fidelity. Here, we assessed the role of DksA in transcription fidelity by monitoring stochastic epigenetic switching in the lac operon (with and without an error-prone transcription slippage sequence), partial phenotypic suppression of a lacZ nonsense allele, as well as monitoring the number of lacI mRNA transcripts produced in the presence and absence of DksA via an operon fusion and single molecule fluorescent in situ hybridization studies. We present data showing that DksA acts to maintain transcription fidelity in vivo and the role of DksA seems to be distinct from that of the GreA and GreB transcription fidelity factors.
DksA是一种辅助转录因子,它与RNA聚合酶相互作用并影响基因表达。根据启动子的不同,DksA可以是转录起始的正调控因子或负调控因子。此外,DksA对转录延伸有重大影响,它能防止转录和复制机器的碰撞,在翻译和转录解偶联时维持转录延伸过程中起关键作用,并且已被证明与转录保真度有关。在这里,我们通过监测乳糖操纵子中的随机表观遗传转换(有或没有易出错的转录滑动序列)、lacZ无义等位基因的部分表型抑制,以及通过操纵子融合和单分子荧光原位杂交研究监测有无DksA时产生的lacI mRNA转录本数量,来评估DksA在转录保真度中的作用。我们提供的数据表明,DksA在体内维持转录保真度,并且DksA的作用似乎与GreA和GreB转录保真因子的作用不同。