Motility gene expression in L. ruminis ATCC25644 vs L. ruminis ATCC27782 during motile and non-motile growth in MRS media
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PROVIDER: PRJNA145677 | ENA |
REPOSITORIES: ENA
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Project description:The genus Lactobacillus contains over 100 different species that were traditionally considered to be uniformly non-motile. However, at least twelve motile species are known to exist in the L. salivarius clade of this genus. Of these, Lactobacillus rumnis is the only motile species that is also autochthonous to the mammalian gastrointestinal tract. The genomes of two L. ruminis strains, ATCC25644 (human isolate, non-motile) and ATCC27782 (bovine isolate, motile) were sequenced and annotated to identify the genes responsible for flagellum biogenesis and chemotaxis in this species. Transcriptome analysis revealed that motility genes were transcribed at a significantly higher level in motile L. ruminis ATCC27782 than in non-motile ATCC25644 during the motile growth phase. In preparation for RNA isolation from L. ruminis ATCC27782 and ATCC25644 cultures in the motile phase, 20 ml aliquots of MRS broth were inoculated (0.25 % inoculum) with a turbid culture of the desired strain. The cultures were incubated anaerobically at 37 °C for 15 hr. In preparation for RNA isolation from L. ruminis ATCC27782 and ATCC25644 cultures in the non-motile growth phase, 20 ml aliquots of MRS broth were inoculated (1 % inoculum) with a turbid culture of the desired strain. The cultures were incubated anaerobically at 37 °C for 16-20 hrs. RNAprotect Bacteria reagent (Qiagen) was used to stabilize gene expression in the target cultures, and total RNA was isolated according to the protocol for enzymatic and mechanical disruption of bacteria as described in the RNAprotect Bacteria Reagent handbook. RNA isolation was completed with the RNeasy Mini kit, and contaminating DNA was removed with the Turbo DNA-free kit (Ambion). An Oligo aCGH/ChIP-on chip hybridization kit (Agilent) was used for hybridization of the labelled cDNA to the microarrays. Probe hybridization took place at 65 °C for 20 hrs with constant rotation (10 rpm). Microarrays were scanned using the Agilent Microarray Scanner System (G2505B) and the scanned files were converted to data files with Feature Extraction software (Agilent). Three microarrays, (three biological replicates) were used to examine gene transcription during the motile growth phase. One microarray was used to examine gene transcription during the non-motile growth phase.
2012-07-09 | E-GEOD-31556 | biostudies-arrayexpress
Project description:The genus Lactobacillus contains over 100 different species that were traditionally considered to be uniformly non-motile. However, at least twelve motile species are known to exist in the L. salivarius clade of this genus. Of these, Lactobacillus rumnis is the only motile species that is also autochthonous to the mammalian gastrointestinal tract. The genomes of two L. ruminis strains, ATCC25644 (human isolate, non-motile) and ATCC27782 (bovine isolate, motile) were sequenced and annotated to identify the genes responsible for flagellum biogenesis and chemotaxis in this species. Transcriptome analysis revealed that motility genes were transcribed at a significantly higher level in motile L. ruminis ATCC27782 than in non-motile ATCC25644 during the motile growth phase.
2012-07-10 | GSE31556 | GEO
Project description:Expression and use of motility genes is a potentially beneficial but costly process in bacteria. Interestingly, many isolate strains of Escherichia coli possess motility genes but have lost the ability to activate them in conditions in which motile cells are advantageous, raising the question of how they respond to these situations. Through transcriptome profiling of strains in the E. coli single-gene knockout Keio collection, we noticed drastic upregulation of motility genes in many of the deletion strains as compared to its typically non-motile parent strain (BW25113). This switch to a motile phenotype is not a direct consequence of the genes deleted, but is instead due to a variety of secondary mutations that increase the synthesis of the major motility regulator, FlhDC. We found that a phenotypic switch to motility at a population level can be induced in non-motile E. coli strains by incubation in non-shaking liquid medium overnight but not in shaking media. Individual isolates after the overnight incubation acquired distinct mutations upstream of the flhDC operon, including different insertion sequence (IS) elements and, to a lesser extent, point mutations. The rapid sweep in the non-shaking population shows that non-motile strains without existing regulatory mechanisms can quickly adapt to a motile lifestyle by quickly acquired genetic changes.
2019-07-11 | GSE129161 | GEO
Project description:These series of experiments compares the expression profile of the motility variants of E.coli MG1655 ( Motile and NonMotile isolates) to an isogenic E.coli MG1655 strain in which the IS5 upstream of flhDC has been deleted. The expression profiles of genes in the E.coli MG1655 motile isolate and E.coli MG1655 Non_Motile isolate also compared.
2010-06-09 | E-GEOD-2694 | biostudies-arrayexpress
Project description:These series of experiments compares the expression profile of the motility variants of E.coli MG1655 ( Motile and NonMotile isolates) to an isogenic E.coli MG1655 strain in which the IS5 upstream of flhDC has been deleted. The expression profiles of genes in the E.coli MG1655 motile isolate and E.coli MG1655 Non_Motile isolate also compared. Keywords: parallel sample
2005-06-02 | GSE2694 | GEO
Project description:Bacterial motility shows a strong evolvable feature depending on the environment. Hyper-motile E. coli could be isolated by evolving non-motile E. coli due to the mutations that enhanced transcriptional expression of the master regulator of the flagellum biosynthesis, FlhDC. These hyper-motile isolates showed reduced growth fitness but with the molecular mechanisms unrevealed. Here we obtained a novel type of hyper-motile isolates by evolving a weakly-motile E. coli K12 strain on the soft agar plates. These isolates carried high accumulated FlhDC proteins and they shared one single point mutation of ClpXV78F. The V78F affected the ATP binding to ClpX via steric repulsive effect and the mutated ClpXP protease lost most of its ability to degraded FlhDC and some other of its known targets. The signal tag of FlhDC for ClpXP recognition was also characterized. Intriguingly, in the hyper-motile strains, the highly enhanced expression of the motility genes was accompanied by the reduced expression of stress resistance genes relating to the reduced fitness of these isolates. Hence, ClpX appeared to be a novel and hot locus during the evolution of bacterial motility and the molecular mechanism of the trade-off between motility and growth was proposed for the first time.
2021-11-15 | GSE165151 | GEO
Project description:To cope with sudden changes in their environment, bacteria can use a bet-hedging strategy by dividing the population into cells with different properties. This so called bimodal or bistable cellular differentiation is generally controlled by positive feedback regulation of transcriptional activators. Due to continued turnover of the cytoplasmic content, it is difficult for these activators to reach an activation threshold concentration when cells are growing exponentially. This is one reason why bimodal differentiation is primarily observed from the onset of the stationary phase when exponential growth ceases. An exception is the bimodal induction of motility in Bacillus subtilis, which occurs early during exponential growth. Several mechanisms have been put forward to explain this, including double negative-feedback regulation and the stability of the mRNA molecules involved. In this study, we used fluorescence-assisted cell sorting to compare the transcriptome of motile and non-motile cells and noted that expression of ribosomal genes is lower in motile cells. This was confirmed using an unstable GFP reporter fused to the strong ribosomal rpsD promoter. We propose that the reduction in ribosomal gene expression in motile cells is the result of a diversion of cellular resources to the synthesis of the chemotaxis and motility systems. In agreement, single-cell microscopic analysis indicates that motile cells are slightly shorter than non-motile cells, an indication of slower growth. We speculate that this growth rate reduction can contribute to the bimodal induction of motility during exponential growth.
2021-01-20 | GSE164348 | GEO
Project description:The challenge of predicting which patients with breast cancer will develop metastases leads to the overtreatment of patients with benign disease and to the inadequate treatment of the aggressive cancers. Here, we report the development and testing of a microfluidic assay that quantifies the abundance and proliferation of migratory cells in breast-cancer specimens, for the assessment of their metastatic propensity and for the rapid screening of potential antimetastatic therapeutics. On the basis of the key roles of cell motility and proliferation in cancer metastasis, the device accurately predicts the metastatic potential of breast-cancer cell lines and of patient-derived xenografts. Compared to unsorted cancer cells, highly motile cells isolated by the device exhibited similar tumourigenic potential but markedly increased metastatic propensity in vivo. RNA sequencing of the highly motile cells revealed an enrichment of motility-related and survival-related genes. The approach might be developed into a companion assay for the prediction of metastasis in patients and for the selection of effective therapeutic regimens.
2019-05-08 | GSE128313 | GEO
Project description:Bacterial motility shows a strong evolvable feature depending on the environment. Hyper-motile E. coli could be isolated by evolving non-motile E. coli due to the mutations that enhanced transcriptional expression of the master regulator of the flagellum biosynthesis, FlhDC. These hyper-motile isolates showed reduced growth fitness but with the molecular mechanisms unrevealed. Here we obtained a novel type of hyper-motile isolates by evolving a weakly-motile E. coli K12 strain on the soft agar plates. These isolates carried high accumulated FlhDC proteins and they shared one single point mutation of ClpXV78F. The V78F affected the ATP binding to ClpX via steric repulsive effect and the mutated ClpXP protease lost most of its ability to degraded FlhDC and some other of its known targets. The signal tag of FlhDC for ClpXP recognition was also characterized. Intriguingly, in the hyper-motile strains, the highly enhanced expression of the motility genes was accompanied by the reduced expression of stress resistance genes relating to the reduced fitness of these isolates. Hence, ClpX appeared to be a novel and hot locus during the evolution of bacterial motility and the molecular mechanism of the trade-off between motility and growth was proposed for the first time.
2021-11-25 | PXD023645 | Pride
Project description:Vibrio cholerae is highly motile by the action of a single polar flagellum. The loss of motility reduces the infectivity of V. cholerae, demonstrating that motility is an important virulence factor. FlrC is the sigma-54-dependent positive regulator of flagellar genes. Recently, the genes VC2206 (flgP) and VC2207 (flgO) were identified as being regulated by FlrC by microarray analysis of an flrC mutant. FlgP is reported to be an outer membrane lipoprotein required for motility that functions as a colonization factor. The study reported here focuses on the characterization of flgO, the first gene in the flgOP operon. We show FlgO/P are important for motility, as these mutants have reduced motility phenotypes. The flgO/P mutant populations display fewer motile cells as well as reduced numbers of flagellated cells. The flagella produced by the flgO/P mutant strains are shorter in length than the WT flagella, which can be restored by inhibiting rotation of the flagellum. FlgO is an outer membrane protein that localizes throughout the membrane and not at the flagellar pole. Although FlgO/P do not specifically localize to the flagellum, they are required for flagellar stability. Due to the nature of these motility defects, we established that the flagellum is not sufficient for adherence, rather, motility is the essential factor required for attachment and thus colonization by V. cholerae O1 of the classical biotype. This study reveals a novel mechanism for which the OMPs FlgO and FlgP function in motility to mediate flagellar stability and influence attachment and colonization. Vibrio cholerae O395 vs. rpoN mutant
2010-05-16 | E-GEOD-17144 | biostudies-arrayexpress