title: Transcriptome-based analysis of the Pantoea stewartii quorum-sensing regulon and identification of EsaR direct targets.
abstract: Pantoea stewartii subsp. stewartii is a proteobacterium that causes Stewart's wilt disease in corn plants. The bacteria form a biofilm in the xylem of infected plants and produce capsule that blocks water transport, eventually causing wilt. At low cell densities, the quorum-sensing (QS) regulatory protein EsaR is known to directly repress expression of esaR itself as well as the genes for the capsular synthesis operon transcription regulator, rcsA, and a 2,5-diketogluconate reductase, dkgA. It simultaneously directly activates expression of genes for a putative small RNA, esaS, the glycerol utilization operon, glpFKX, and another transcriptional regulator, lrhA. At high bacterial cell densities, all of this regulation is relieved when EsaR binds an acylated homoserine lactone signal, which is synthesized constitutively over growth. QS-dependent gene expression is critical for the establishment of disease in the plant. However, the identity of the full set of genes controlled by EsaR/QS is unknown. A proteomic approach previously identified around 30 proteins in the QS regulon. In this study, a whole-transcriptome, next-generation sequencing analysis of rRNA-depleted RNA from QS-proficient and -deficient P. stewartii strains was performed to identify additional targets of EsaR. EsaR-dependent transcriptional regulation of a subset of differentially expressed genes was confirmed by quantitative reverse transcription-PCR (qRT-PCR). Electrophoretic mobility shift assays demonstrated that EsaR directly bound 10 newly identified target promoters. Overall, the QS regulon of P. stewartii orchestrates three major physiological responses: capsule and cell envelope biosynthesis, surface motility and adhesion, and stress response.
authors.affiliation: Department of Biological Sciences, Virginia Tech, Blacksburg, Virginia, USA.
authors.forename: Revathy
authors.initials: R
authors.lastname: Ramachandran
authors.affiliation: Department of Biological Sciences, Virginia Tech, Blacksburg, Virginia, USA.
authors.forename: Alison Kernell
authors.initials: AK
authors.lastname: Burke
authors.affiliation: University of Georgia, Georgia Advanced Computing Resource Center and Institute of Bioinformatics, Athens, Georgia, USA.
authors.forename: Guy
authors.initials: G
authors.lastname: Cormier
authors.affiliation: Department of Biological Sciences, Virginia Tech, Blacksburg, Virginia, USA.
authors.forename: Roderick V
authors.initials: RV
authors.lastname: Jensen
authors.affiliation: Department of Biological Sciences, Virginia Tech, Blacksburg, Virginia, USA ams@vt.edu.
authors.forename: Ann M
authors.initials: AM
authors.lastname: Stevens
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism*; D004269:DNA, Bacterial / Q000378:metabolism; D024202:Electrophoretic Mobility Shift Assay; D020869:Gene Expression Profiling; D053263:Gene Regulatory Networks; D059014:High-Throughput Nucleotide Sequencing; D020636:Pantoea / Q000235:genetics / Q000502:physiology*; D010935:Plant Diseases / Q000382:microbiology; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D053038:Quorum Sensing; D060888:Real-Time Polymerase Chain Reaction; D018085:Regulon; D014157:Transcription Factors / Q000378:metabolism*; D003313:Zea mays / Q000382:microbiology
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial; C095459:EsaR protein, Pantoea stewartii; D014157:Transcription Factors