Transcription initiation involves the conversion from closed promoter complexes, comprising RNA polymerase (RNAP) and double-stranded promoter DNA, to open complexes, in which the enzyme is able to access the DNA template in a single-stranded form. The complex between bacterial RNAP and its major variant sigma factor σ54 remains as a closed complex until ATP hydrolysis-dependent remodeling by activator proteins occurs. This remodeling facilitates DNA melting and allows the transition to the open complex. Here we present cryoelectron microscopy reconstructions of bacterial RNAP in complex with σ54 alone, and of RNAP-σ54 with an AAA+ activator. Together with photo-crosslinking data that establish the location of promoter DNA within the complexes, we explain why the RNAP-σ54 closed complex is unable to access the DNA template and propose how the structural changes induced by activator binding can initiate conformational changes that ultimately result in formation of the open complex. © 2008 Elsevier Inc. All rights reserved.
Bose, D., Pape, T., Burrows, P. C., Rappas, M., Wigneshweraraj, S. R., Buck, M., & Zhang, X. (2008). Organization of an Activator-Bound RNA Polymerase Holoenzyme. Molecular Cell, 32(3), 337–346. https://doi.org/10.1016/j.molcel.2008.09.015