The transcriptional activator LdtR from 'Candidatus Liberibacter asiaticus' mediates osmotic stress tolerance

• Published in: PLoS pathogens 2014-04, Vol.10 (4), p.e1004101-e1004101
• Main Authors: Pagliai, Fernando A, Gardner, Christopher L, Bojilova, Lora, Sarnegrim, Amanda, Tamayo, Cheila, Potts, Anastasia H, Teplitski, Max, Folimonova, Svetlana Y, Gonzalez, Claudio F, Lorca, Graciela L
• Format: Article
• Language: English
• Subjects: Bacteria - genetics; Bacteria - metabolism; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Bacteriology; Binding sites; Biology and Life Sciences; Citrus - microbiology; Diamonds; Genes; Genetic aspects; Genetic research; Genetic transcription; Genomes; Health aspects; Host-parasite relationships; Ligands; Medicine and Health Sciences; Methods; Microbiological research; Osmotic Pressure; Plant Diseases - genetics; Plant Diseases - microbiology; Proteins; Proteobacteria; Studies; Trans-Activators - genetics; Trans-Activators - metabolism; Transcription factors; Trees
• ISSN: 1553-7374; 1553-7366; 1553-7374
• DOI: 10.1371/journal.ppat.1004101

The causal agent of Huanglongbing disease, 'Candidatus Liberibacter asiaticus', is a non-culturable, gram negative, phloem-limited α-proteobacterium. Current methods to control the spread of this disease are still limited to the removal and destruction of infected trees. In this study, we identified and characterized a regulon from 'Ca. L. asiaticus' involved in cell wall remodeling, that contains a member of the MarR family of transcriptional regulators (ldtR), and a predicted L,D-transpeptidase (ldtP). In Sinorhizobium meliloti, mutation of ldtR resulted in morphological changes (shortened rod-type phenotype) and reduced tolerance to osmotic stress. A biochemical approach was taken to identify small molecules that modulate LdtR activity. The LdtR ligands identified by thermal shift assays were validated using DNA binding methods. The biological impact of LdtR inactivation by the small molecules was then examined in Sinorhizobium meliloti and Liberibacter crescens, where a shortened-rod phenotype was induced by growth in presence of the ligands. A new method was also developed to examine the effects of small molecules on the viability of 'Ca. Liberibacter asiaticus', using shoots from HLB-infected orange trees. Decreased expression of ldtRLas and ldtPLas was observed in samples taken from HLB-infected shoots after 6 h of incubation with the LdtR ligands. These results provide strong proof of concept for the use of small molecules that target LdtR, as a potential treatment option for Huanglongbing disease.