Here we describe approaches and methods to assaying in vitro the major variant bacterial sigma factor, Sigma 54 (σ54), in a purified system. We include the complete transcription system, binding interactions between σ54 and its activators, as well as the self-assembly and the critical ATPase activity of the cognate activators which serve to remodel the closed promoter complexes. We also present in vivo methodologies that are used to study the impact of physiological processes, metabolic states, global signalling networks, and cellular architecture on the control of σ54-dependent gene expression.
|Title of host publication||Bacterial Transcriptional Control: methods and protocols|
|Editors||Irina Artsimovitch, Thomas J Santangelo|
|Place of Publication||New York|
|Publication status||Published - 28 Jan 2015|
|Name||Methods in Molecular Biology|
|Publisher||Springer New York|
Buck, M., Engl, C., Joly, N., Jovanovic, G., Lawton, E., McDonald, C., Schumacher, J., Waite, C. J., & Zhang, N. (2015). In vitro and in vivo methodologies for studying the Sigma 54-dependent transcription. In I. Artsimovitch, & T. J. Santangelo (Eds.), Bacterial Transcriptional Control: methods and protocols (Vol. 1276, pp. 53-79). (Methods in Molecular Biology). Springer. https://doi.org/10.1007/978-1-4939-2392-2_4