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利用微流控力学捕获技术对大肠杆菌 RNA 聚合酶的蛋白质-蛋白质相互作用进行作图。

Mapping of protein-protein interactions of E. coli RNA polymerase with microfluidic mechanical trapping.

机构信息

Department of Applied Physics, Stanford University, Stanford, California, United States of America.

Department of Applied Physics, Stanford University, Stanford, California, United States of America; Department of Bioengineering and HHMI, Stanford University, Stanford, California, United States of America.

出版信息

PLoS One. 2014 Mar 18;9(3):e91542. doi: 10.1371/journal.pone.0091542. eCollection 2014.

Abstract

The biophysical details of how transcription factors and other proteins interact with RNA polymerase are of great interest as they represent the nexus of how structure and function interact to regulate gene expression in the cell. We used an in vitro microfluidic approach to map interactions between a set of ninety proteins, over a third of which are transcription factors, and each of the four subunits of E. coli RNA polymerase, and we compared our results to those of previous large-scale studies. We detected interactions between RNA polymerase and transcription factors that earlier high-throughput screens missed; our results suggest that such interactions can occur without DNA mediation more commonly than previously appreciated.

摘要

转录因子和其他蛋白质与 RNA 聚合酶相互作用的生物物理细节非常令人感兴趣,因为它们代表了结构和功能如何相互作用以调节细胞中基因表达的交点。我们使用体外微流控方法来绘制一组 90 种蛋白质(其中超过三分之一是转录因子)与大肠杆菌 RNA 聚合酶的四个亚基之间的相互作用,并将我们的结果与以前的大规模研究进行了比较。我们检测到了 RNA 聚合酶与转录因子之间的相互作用,而早期的高通量筛选却错过了这些相互作用;我们的结果表明,这种相互作用可以在没有 DNA 介导的情况下更频繁地发生,这比以前的认识更为普遍。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/624d/3958368/bb0bf5e5ac06/pone.0091542.g001.jpg

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