Exploring RNA Polymerase Regulation by NMR Spectroscopy

Overview

Journal Sci Rep

Specialty Science

Date 2015 Jun 5

PMID 26043358

Citations 12

Authors

Johanna Drogemuller

Martin Strauss

Kristian Schweimer

Birgitta M Wohrl

Stefan H Knauer

Paul Rosch

Affiliations

Soon will be listed here.

Abstract

RNA synthesis is a central process in all organisms, with RNA polymerase (RNAP) as the key enzyme. Multisubunit RNAPs are evolutionary related and are tightly regulated by a multitude of transcription factors. Although Escherichia coli RNAP has been studied extensively, only little information is available about its dynamics and transient interactions. This information, however, are crucial for the complete understanding of transcription regulation in atomic detail. To study RNAP by NMR spectroscopy we developed a highly efficient procedure for the assembly of active RNAP from separately expressed subunits that allows specific labeling of the individual constituents. We recorded [(1)H,(13)C] correlation spectra of isoleucine, leucine, and valine methyl groups of complete RNAP and the separately labeled β' subunit within reconstituted RNAP. We further produced all RNAP subunits individually, established experiments to determine which RNAP subunit a certain regulator binds to, and identified the β subunit to bind NusE.

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