Issue 6, 2022

Cu(ii)-based DNA labeling identifies the structural link between transcriptional activation and termination in a metalloregulator

Abstract

Understanding the structural and mechanistic details of protein-DNA interactions that lead to cellular defence against toxic metal ions in pathogenic bacteria can lead to new ways of combating their virulence. Herein, we examine the Copper Efflux Regulator (CueR) protein, a transcription factor which interacts with DNA to generate proteins that ameliorate excess free Cu(I). We exploit site directed Cu(II) labeling to measure the conformational changes in DNA as a function of protein and Cu(I) concentration. Unexpectedly, the EPR data indicate that the protein can bend the DNA at high protein concentrations even in the Cu(I)-free state. On the other hand, the bent state of the DNA is accessed at a low protein concentration in the presence of Cu(I). Such bending enables the coordination of the DNA with RNA polymerase. Taken together, the results lead to a structural understanding of how transcription is activated in response to Cu(I) stress and how Cu(I)-free CueR can replace Cu(I)-bound CueR in the protein-DNA complex to terminate transcription. This work also highlights the utility of EPR to measure structural data under conditions that are difficult to access in order to shed light on protein function.

Graphical abstract: Cu(ii)-based DNA labeling identifies the structural link between transcriptional activation and termination in a metalloregulator

Supplementary files

Article information

Article type
Edge Article
Submitted
24 nov 2021
Accepted
16 jan 2022
First published
17 jan 2022
This article is Open Access

All publication charges for this article have been paid for by the Royal Society of Chemistry
Creative Commons BY-NC license

Chem. Sci., 2022,13, 1693-1697

Cu(II)-based DNA labeling identifies the structural link between transcriptional activation and termination in a metalloregulator

J. Casto, A. Mandato, L. Hofmann, I. Yakobov, S. Ghosh, S. Ruthstein and S. Saxena, Chem. Sci., 2022, 13, 1693 DOI: 10.1039/D1SC06563G

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