Borrelia host adaptation Regulator (BadR) regulates rpoS to modulate host adaptation and virulence factors in Borrelia burgdorferi
Ontology highlight
ABSTRACT:
ORGANISM(S): Borreliella burgdorferi B31
PROVIDER: GSE38827 | GEO | 2013/02/22
SECONDARY ACCESSION(S): PRJNA169019
REPOSITORIES: GEO
Similar Datasets
Project description:Our study is the first to demonstrate BadR as a repressor of rpoS and thus facilitates spirocheteM-bM-^@M-^Ys transition back into ticks. BadR binds upstream and represses rpoS in unfed ticks. GLcNA-6P, an abundant nutrient of blood and released by subsequent remodeling of the tick peritrophic membrane, relieves repression of BadR on rpoS facilitating vertebrate host adaptation. BadR regulates chitobiose utilization genes and activates genes critical for spirochetes residence of ticks (lp28-4 genes) badR-deficient strain (Gene BB0693) compared to B31-A3 parental wild-type B. burgdorferi strains Two separate but identically designed microarrays were performed and thus two chips were used. Each chip contained two replicates of B31-A3 WT and badR-deficient strains, and thus the overall array analysis of both chips corresponds to n=4 for both WT and badR-deficient strains. Hierarchical clustering and correlation analysis was used to verify quality of the normalized array data. The expression data from biological replicates 1 and 2 (chip 1 and 2) were both pooled and analyzed individually. We used LIMMA R package to test contrasts between WT and badR-deficient strain (Smyth GK.2004. Stat Appl Genet Mol Biol 3:Article3 .)
2013-02-22 | E-GEOD-38827 | biostudies-arrayexpress
Project description:Lyme disease is a result from an infection by the spirochete Borrelia burgdorferi, and is the leading vector-borne disease in North America. Due to the genomic and proteomic variability of different B. burgdorferi isolates, the study and further comparison of their proteome is key to understand the biology and infectivity of these spirochetes. Mass spectrometry-based proteomics was used to assemble peptide datasets of laboratory isolates B31, MM1, B31-ML23, and the infective isolates B31-5A4, B31-A3, and 297, as well as other public datasets, and is publicly available as the Borrelia PeptideAtlas (http://www.peptideatlas.org/builds/borrelia/). These datasets include information on total proteome, secretome, and membrane proteome of the B. burgdorferi.
2024-09-03 | PXD046281 | Pride
Project description:The RpoS transcription factor of Borrelia burgdorferi is a 'gatekeeper' because it activates genes required for spirochaetes to transition from tick to vertebrate hosts. However, it remains unknown how RpoS becomes repressed to allow the spirochaetes to transition back from the vertebrate host to the tick vector. Here we show that a putative carbohydrate-responsive regulatory protein, designated BadR (Borrelia host adaptation Regulator), is a transcriptional repressor of rpoS. BadR levels are elevated in B. burgdorferi cultures grown under in vitro conditions mimicking unfed-ticks and badR-deficient strains are defective for growth under these same conditions. Microarray and immunoblot analyses of badR-deficient strains showed upregulation of rpoS and other factors important for virulence in vertebrate hosts, as well as downregulation of putative tick-specific determinants (e.g. linear plasmid 28-4 genes). DNA-binding assays revealed BadR binds to upstream regions of rpoS. Site-directed mutations in BadR and the presence of phosphorylated sugars affected BadR's binding to the rpoS promoters. badR-deficient B. burgdorferi were unable to colonize mice. Several putative tick-specific targets have been identified. Our study identified a novel regulator, BadR, and provides a link between nutritional environmental cues utilized by spirochaetes to adaptation to disparate conditions found in the tick and vertebrate hosts.
| S-EPMC4828661 | biostudies-literature
Project description:The RpoN-RpoS alternative sigma factor pathway is essential for key adaptive responses by Borrelia burgdorferi, particularly those involved in the infection of a mammalian host. A putative response regulator, Rrp2, ostensibly is required for activation of the RpoN-dependent transcription of rpoS. However, questions remain regarding the extent to which the three major constituents of this pathway (Rrp2, RpoN, and RpoS) act interdependently. To assess the functional interplay between Rrp2, RpoN, and RpoS, we employed microarray analyses to compare gene expression levels in rrp2, rpoN, or rpoS mutants of parental strain 297. We identified 98 genes that were similarly regulated by Rrp2, RpoN and RpoS, and an additional 47 genes were determined to be likely regulated by this pathway. The substantial overlap between genes regulated by RpoS and RpoN provides compelling evidence that these two alternative sigma factors form a congruous pathway and that RpoN regulates B. burgdorferi gene expression through RpoS. Although several known B. burgdorferi virulence determinants were regulated by the RpoN-RpoS pathway, a defined function has yet to be ascribed to most of the genes substantially regulated by Rrp2, RpoN and RpoS, therefore providing additional avenues for future research into the genetic determinants contributing to virulence expression by B. burgdorferi. Keywords: gene profiling
2008-09-09 | GSE11284 | GEO
Project description:A collection of 1145 clones from an EST project on female tick salivary gland genes was hybridized on glass slides to RNA extracted from several feeding stages of adult female tick salivary glands, including unfed and replete, and from adult male ticks, either unfed or fed in the presence or absence of female ticks. In the female ticks, the early fed (<50 mg) and partially fed (30-200 mg) groups were very similar. The fast feeding (350-500 mg) and replete ticks were similar to each other, but different from the partially fed. The unfed ticks were more similar to the fast feeding – replete groups than the early fed-partially fed groups. In the males, there were differences between the males fed in the presence or absence of females, but overall, these groups were very similar. The unfed ticks were significantly different from the fed ticks. Males showed clear differences with females in expression, as well. The unfed females had high levels of genes involved in protein synthesis, while genes possibly involved in survival on the host, such as anticoagulants, seemed to be most expressed in the early and partially fed states. By contrast, in the males, the protein synthesis genes were expressed more in all three groups, while the putative secreted genes for survival were expressed less. Keywords: time course, effect of feeding, sex, effect of presence of females
2006-06-08 | GSE5010 | GEO
Project description:Lyme disease spirochetes must induce RpoS-dependent genes during tick feeding to prepare for host infection. Previous work in our lab identified bbd18 as a negative regulator of RpoS, but inactivation of bbd18 in wild-type spirochetes was never achieved. In the current study we generated an inducible bbd18 gene at the endogenous plasmid locus and demonstrated the essential nature of BBD18 for viability of wild-type spirochetes in vitro and at a unique point in vivo. Transcriptomic analyses demonstrated global induction of RpoS and RpoS-dependent genes following BBD18 depletion, culminating in spirochete lysis. Plasmid prophage genes were also induced and phage particles were detected in lysed culture supernatants, suggesting that RpoS regulates phage lysis-lysogeny decisions. The absolute requirement for BBD18 persisted following displacement of the entire set of cp32 plasmid prophages but could be circumvented by deletion of rpoS. This is the first report of a mechanistic link between endogenous transducing prophages and the RpoS-dependent adaptive response of the Lyme disease spirochete.
2022-12-12 | GSE207123 | GEO
Project description:A collection of 1145 clones from an EST project on female tick salivary gland genes was hybridized on glass slides to RNA extracted from several feeding stages of adult female tick salivary glands, including unfed and replete, and from adult male ticks, either unfed or fed in the presence or absence of female ticks. In the female ticks, the early fed (<50 mg) and partially fed (30-200 mg) groups were very similar. The fast feeding (350-500 mg) and replete ticks were similar to each other, but different from the partially fed. The unfed ticks were more similar to the fast feeding â replete groups than the early fed-partially fed groups. In the males, there were differences between the males fed in the presence or absence of females, but overall, these groups were very similar. The unfed ticks were significantly different from the fed ticks. Males showed clear differences with females in expression, as well. The unfed females had high levels of genes involved in protein synthesis, while genes possibly involved in survival on the host, such as anticoagulants, seemed to be most expressed in the early and partially fed states. By contrast, in the males, the protein synthesis genes were expressed more in all three groups, while the putative secreted genes for survival were expressed less. Keywords: time course, effect of feeding, sex, effect of presence of females All samples were compared to the partially fed females. Females consisted of five groups: unfed, early fed, partially fed, fast feeding and replete. Four or five biological replicates were done of each, with the dyes used in both possible ways. In the males, three groups were used: unfed, feeding in the presence of females, and feeding in the absence of females. Two biological replicates were done of the feeding males, and one of extracts was hybridized twice for the males fed in the presence of females. Unfed males used one RNA sample, extracted from a large pool of ticks.
2010-06-11 | E-GEOD-5010 | biostudies-arrayexpress
Project description:The RpoN-RpoS alternative sigma factor pathway is essential for key adaptive responses by Borrelia burgdorferi, particularly those involved in the infection of a mammalian host. A putative response regulator, Rrp2, ostensibly is required for activation of the RpoN-dependent transcription of rpoS. However, questions remain regarding the extent to which the three major constituents of this pathway (Rrp2, RpoN, and RpoS) act interdependently. To assess the functional interplay between Rrp2, RpoN, and RpoS, we employed microarray analyses to compare gene expression levels in rrp2, rpoN, or rpoS mutants of parental strain 297. We identified 98 genes that were similarly regulated by Rrp2, RpoN and RpoS, and an additional 47 genes were determined to be likely regulated by this pathway. The substantial overlap between genes regulated by RpoS and RpoN provides compelling evidence that these two alternative sigma factors form a congruous pathway and that RpoN regulates B. burgdorferi gene expression through RpoS. Although several known B. burgdorferi virulence determinants were regulated by the RpoN-RpoS pathway, a defined function has yet to be ascribed to most of the genes substantially regulated by Rrp2, RpoN and RpoS, therefore providing additional avenues for future research into the genetic determinants contributing to virulence expression by B. burgdorferi. Keywords: gene profiling 297 (wt) vs rpoS mutant: 6 slides, 12 hybridizations (including dyeswaps), three biological replicates 297 (wt) vs rpoN mutant: 5 slides, 10 hybridizations (including dyeswaps), three biological replicates 297 (wt) vs rrp2 mutant: 6 slides, 12 hybridizations (including dyeswaps), three biological replicates 5A18 (wt) vs XY424.3 (rrp2 mutant): 5 slides, 10 hybridizations (including dyeswaps), three biological replicates 297 (wt) vs rpoS mutant: 6 slides, 12 hybridizations (including dyeswaps), three biological replicates 297 (wt) vs rpoN mutant: 5 slides, 10 hybridizations (including dyeswaps), three biological replicates 297 (wt) vs rrp2 mutant: 6 slides, 12 hybridizations (including dyeswaps), three biological replicates 5A18 (wt) vs XY424.3 (rrp2 mutant): 5 slides, 10 hybridizations (including dyeswaps), three biological replicates
2008-09-09 | E-GEOD-11284 | biostudies-arrayexpress
Project description:We developed a new simple method to assess the composition of proteins in the Ornithodoros moubata saliva. To collect naturally expectorated saliva from the ticks, we employed an artificial membrane feeding technique using a simple, chemically defined diet and submitted the collected native saliva samples for liquid chromatography-mass spectrometry (LC-MS) analysis. These experiments were conducted with groups of uninfected ticks as well as with B. duttonii infected ticks. The ticks exhibited a fair feeding response to the tested diet, engorgement rates reaching between 60-100% of ticks per feeding chamber. LC-MS analysis identified a total of 17 and 15 proteins in saliva samples from the uninfected and infected ticks respectively. Importantly, the analysis was sensitive enough to detect up to 9 different proteins in the saliva containing diet upon which as few as 6 nymphal ticks fed. Some of the proteins identified are well known for their immunomodulatory activity in a vertebrate host, whereas others were structural or “housekeeping” proteins and their finding in the naturally expectorated saliva confirms that they can be secreted and having some roles at the tick-host interface. Most notably, some of the proteins that have long been suspected as important for vector-pathogen interactions of Borrelia spirochetes were detected only in the samples from infected ticks, suggesting that their expression was altered by the persistent colonization of the tick’s salivary glands by spirochetes. The simple method described herein is an important addition to the toolbox available to study the vector-host-pathogen interactions in the rapidly feeding soft ticks.
2023-01-10 | PXD038824 | Pride
Project description:Understanding the molecular basis of how the tick adapts to feed on different animal hosts is central to understanding tick and tick-borne disease (TBD) epidemiology. Tick adaptation to feed on vertebrate hosts is regulated by tick secretion of multiple tick saliva proteins (TSPs) and other molecules that regulate tick feeding. This study was initiated to determine if ticks such as Ixodes scapularis and Amblyomma americanum that are adapted to feed on multiple hosts utilized the same sets of proteins to accomplish feeding on all hosts. Our data suggest that ticks of the same species differentially express proteins when feeding on diffent hosts. SDS-PAGE and silver staining analysis revealed unique protein eletrophoretic profile in saliva of Ixodes scapularis and Amblyomma americanum that were stimulated to start feeding on different hosts: rabbits, humans, and dogs. LC-MS/MS sequencing and pairwise analysis of proteins in saliva of I. scapularis and A. americanum ticks that were non-stimulated and those that were stimulated to feed on rabbits, dogs, or humans identified TSPs that were unique to each treatment and those that were common. Overal, we identified a total of 276 and 340 non-redundant I. scapularis and A. americanum TSPs, which we have classified into 28 functional classes that include secreted conserved proteins (unknown functions), proteinase inhibitors, lipocalins, extracellular matrix/cell adhesion, heme/iron metabolism, signal transduction and immunity-related proteins being the most predominant in saliva of unfed ticks. With exception of Rhipicephalus microplus, anti-tick vaccine research relies on feeding lab animals. Data here suggest that lab animal data could result in prioritizing irrelevant targets as some tick genes are unique to ticks fed on lab animals. This study provides the platform that could be utilized to identify relevant target anti-tick vaccine antigens, and will facilitate early stage tick feeding research.
2022-02-28 | PXD007712 | Pride