Study of RNA polymerase dynamics during elongation with cryo-electron tomography

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With this application we would like to study the RNA polymerase dynamics during elongation with cryo-electron tomography. We aim to visualize the organization, interactions and conformation of the RNA polymerase and associated macromolecules during elongation. We would like to perform this analysis in yeast and Mycoplasma cells, since the eukaryotic and bacterial systems provide complementary information. The cryo-electron tomograms should visualize the transcription natively in the cellular context. We anticipate achieving a resolution sufficient to mechanistically explain the polymerase conformational states, as well as the interactions of the enzyme with other factors during elongation.