Borrelia burgdorferi PlzA is a cyclic-di-GMP dependent DNA and RNA binding protein

Authors

Nerina Jusufovic, University of Kentucky College of Medicine
Andrew C. Krusenstjerna, University of Kentucky College of Medicine
Christina R. Savage, University of Kentucky College of Medicine
Timothy C. Saylor, University of Kentucky College of Medicine
Catherine A. Brissette, School of Medicine & Health Sciences
Wolfram R. Zückert, University of Kansas School of Medicine
Paula Schlax, Bates CollegeFollow
Md A. Motaleb, The Brody School of Medicine
Brian Stevenson, University of Kentucky College of Medicine

Publication Title

Molecular Microbiology

Document Type

Article

Department or Program

Chemistry

Second Department or Program

Biochemistry

Publication Date

3-2024

Keywords

Borrelia, cyclic-di-GMP, DNA-binding protein, Lyme disease

Abstract

The PilZ domain-containing protein, PlzA, is the only known cyclic di-GMP binding protein encoded by all Lyme disease spirochetes. PlzA has been implicated in the regulation of many borrelial processes, but the effector mechanism of PlzA was not previously known. Here, we report that PlzA can bind DNA and RNA and that nucleic acid binding requires c-di-GMP, with the affinity of PlzA for nucleic acids increasing as concentrations of c-di-GMP were increased. A mutant PlzA that is incapable of binding c-di-GMP did not bind to any tested nucleic acids. We also determined that PlzA interacts predominantly with the major groove of DNA and that sequence length and G–C content play a role in DNA binding affinity. PlzA is a dual-domain protein with a PilZ-like N-terminal domain linked to a canonical C-terminal PilZ domain. Dissection of the domains demonstrated that the separated N-terminal domain bound nucleic acids independently of c-di-GMP. The C-terminal domain, which includes the c-di-GMP binding motifs, did not bind nucleic acids under any tested conditions. Our data are supported by computational docking, which predicts that c-di-GMP binding at the C-terminal domain stabilizes the overall protein structure and facilitates PlzA-DNA interactions via residues in the N-terminal domain. Based on our data, we propose that levels of c-di-GMP during the various stages of the enzootic life cycle direct PlzA binding to regulatory targets.

Comments

Original version is available from the publisher at: https://doi.org/10.1111/mmi.15254

Recommended Citation

Jusufovic, N., Krusenstjerna, A. C., Savage, C. R., Saylor, T. C., Brissette, C. A., Zückert, W. R., Schlax, P. J., Motaleb, M. A., & Stevenson, B. (2024). Borrelia burgdorferi PlzA is a cyclic-di-GMP dependent DNA and RNA binding protein. Molecular Microbiology. https://doi.org/10.1111/mmi.15254

PubMed ID

38527857

Copyright Note

Copyright © 2024 The Authors. Molecular Microbiology published by John Wiley & Sons Ltd.

Required Publisher's Statement

This is an open access article under the terms of the CC-BY-NC license, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes.