Zinc is an inhibitor of the LdtR transcriptional activator

PLoS One. 2018 Apr 10;13(4):e0195746. doi: 10.1371/journal.pone.0195746. eCollection 2018.

Abstract

LdtR is a master regulator of gene expression in Liberibacter asiaticus, one of the causative agents of citrus greening disease. LdtR belongs to the MarR-family of transcriptional regulators and it has been linked to the regulation of more than 180 genes in Liberibacter species, most of them gathered in the following Clusters of Orthologous Groups: cell motility, cell wall envelope, energy production, and transcription. Our previous transcriptomic evidence suggested that LdtR is directly involved in the modulation of the zinc uptake system genes (znu) in the closely related L. crescens. In this report, we show that LdtR is involved in the regulation of one of the two encoded zinc uptake mechanisms in L. asiaticus, named znu2. We also show that LdtR binds zinc with higher affinity than benzbromarone, a synthetic effector inhibitory molecule, resulting in the disruption of the LdtR:promoter interactions. Using site-directed mutagenesis, electrophoretic mobility shift assays (EMSAs), and isothermal titration calorimetry, we identified that residues C28 and T43 in LdtR, located in close proximity to the Benz1 pocket, are involved in the interaction with zinc. These results provided new evidence of a high-affinity effector molecule targeting a key player in L. asiaticus' physiology and complemented our previous findings about the mechanisms of signal transduction in members of the MarR-family.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / antagonists & inhibitors*
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Citrus / microbiology*
  • Gene Expression Regulation, Bacterial / drug effects*
  • Plant Diseases / microbiology*
  • Rhizobiaceae / drug effects
  • Rhizobiaceae / physiology*
  • Transcriptional Activation / drug effects*
  • Zinc / pharmacology*

Substances

  • Bacterial Proteins
  • Zinc

Grants and funding

This work is supported by the Specialty Crop Research Initiative (Award number 2015-70016-23029 to G.L.L. and C.F.G), and the Plant Biotic Interactions Program (Award number 2017-03060 to C.F.G.) from the USDA National Institute of Food and Agriculture, http://nifa.usda.gov/.