Biology subjects

Mauffrey, F.

Publications and source records attributed to Mauffrey, F..

3 recordsLinked to original sources

bioRxiv2019

ScmR, a global regulator of gene expression, quorum sensing, pH homeostasis, and virulence in Burkholderia thailandensis

The nonpathogenic soil saprophyte Burkholderia thailandensis is a member of the Burkholderia pseudomallei-thailandensis-mallei (Bptm) group, which also comprises the closely related human pathogens Burkholderia pseudomallei and Burkholderia mallei responsible for the diseases melioidosis and glanders, respectively. ScmR, a recently identified LysR-type transcriptional regulator (LTTR) in B. thailandensis acts as a global transcriptional regulator throughout the stationary phase, and modulates the production of a wide range of secondary metabolites, including N-acyl-L-homoserine lactones (AHLs) and 4-hydroxy-3-methyl-2-alkylquinoline (HMAQ), virulence in the model host Caenorhabditis elegans, as well as several quorum sensing (QS)-dependent phenotypes. We have investigated the role of ScmR in B. thailandensis strain E264 during the exponential phase. We used RNA-Sequencing (RNA-Seq) transcriptomic analyses to identify the ScmR regulon, which was compared to the QS-controlled regulon, showing a considerable overlap between the ScmR-regulated genes and those controlled by QS. We characterized several genes modulated by ScmR, using quantitative reverse transcription-PCR (qRT-PCR) or mini-CTX-lux transcriptional reporters, including the oxalate biosynthetic gene obc1 required for pH homeostasis, the orphan LuxR-type transcriptional regulator BtaR5-encoding gene, the bsa (Burkholderia secretion apparatus) type III secretion system (T3SS) genes essential for both B. pseudomallei and B. mallei pathogenicity, as well as the scmR gene itself. We confirmed that the transcription of scmR is under QS control, presumably ensuring fine-tuned modulation of gene expression. Finally, we demonstrate that ScmR influences virulence using the fruit fly model host Drosophila melanogaster. We conclude that ScmR represents a central component of the B. thailandensis QS regulatory network. ImportanceCoordination of the expression of genes associated with bacterial virulence and environmental adaptation is often dependent on quorum sensing (QS). The QS circuitry of the nonpathogenic bacterium Burkholderia thailandensis, which is widely used as a model system for the study of the human pathogen Burkholderia pseudomallei, is complex. We found that the recently identified LysR-type transcriptional regulator (LTTR), ScmR, which is highly conserved and involved in the control of virulence/survival factors in the Burkholderia genus, is a global regulator mediating gene expression through the multiple QS systems coexisting in B. thailandensis, as well as independently of QS. We conclude that ScmR represents a key QS modulatory network element, ensuring tight regulation of the transcription of QS-controlled genes, particularly those required for acclimatization to the environment.

microbiology↗

bioRxiv2019

Impact of NaCl, nitrate and temperature on the microbial community of a methanol-fed, denitrifying marine biofilm

BackgroundThe biofilm of a continuous, methanol-fed, fluidized denitrification system that treated a marine effluent at the Montreal Biodome is composed of a multi-species microbial community, among which Hyphomicrobium nitrativorans NL23 and Methylophaga nitratireducenticrescens JAM1 are the principal bacteria involved in the denitrifying activities. To assess its resilience to environmental changes, the biofilm taken from the denitrification system was cultured at laboratory scale in artificial seawater (ASW) under anoxic conditions and exposed to a range of specific physico-chemical parameters. We previously showed that the seawater formulation and the NaCl concentrations had a strong impact on the H. nitrativorans NL23 population, with its displacement by a new denitrifier, M. nitratireducenticrescens GP59. Here, we report the impact of these cultures conditions on the dynamics of the overall microbial community of the denitrifying biofilm.\n\nMethodsThe original biofilm (OB) taken from the denitrification system was acclimated for five weeks in ASW under anoxic conditions with a range of NaCl concentrations, and with four combinations of nitrate concentrations and temperatures. The OB was also acclimated to the commercial Instant Ocean seawater medium (IO). The bacterial diversity of the biofilm cultures and the OB was determined by 16S ribosomal RNA amplicon metagenome sequencing. Culture-dependent approach was used to isolate other denitrifying bacteria from the biofilm cultures. The metatranscriptomes of some of the biofilm cultures were derived, along with the transcriptomes of planktonic pure cultures of H. nitrativorans NL23 and M. nitratireducentricrescens GP59 cultivated under denitrifying conditions.\n\nResultsThe 16S metagenomic data revealed very high proportions of M. nitratireducenticrescens in the biofilm cultures. H. nitrativorans NL23 was found in high proportion in the OB, both was absent in the biofilm cultures with 2.75% NaCl in the ASW medium. It was found however in low proportions in the biofilm cultures with 0 to 1% NaCl in the ASW medium and in the IO biofilm cultures. Emergence of Marinicella spp. occurred in these biofilm cultures. Denitrifying bacterial isolates affiliated to Marinobacter spp. and Paracoccus spp. were isolated. Up regulation of the denitrification genes in strains GP59 and NL23 occurred in the biofilm cultures compared to the planktonic pure cultures. Denitrifying bacteria affiliated to the Stappia spp. were metabolically active in the biofilm cultures.\n\nConclusionsThese results illustrate the dynamics of the microbial community in the denitrifying biofilm cultures in adapting to different environmental conditions. The NaCl concentration is an important factor affecting the microbial community in the biofilm cultures. Up regulation of the denitrification genes in strain GP59 and strain NL23 in the biofilm cultures suggests different mechanisms of regulation of the denitrification pathway in the biofilm compared to the planktonic pure cultures. Other denitrifying heterotrophic bacteria are present in low proportions in the biofilm, suggesting that the biofilm has the potential to adapt to heterotrophic, non-methylotrophic environments.

microbiology↗

bioRxiv2017

Denitrifying metabolism of the methylotrophic marine bacterium Methylophaga nitratireducenticrescens strain JAM1

Methylophaga nitratireducenticrescens strain JAM1 is a methylotrophic, marine bacterium that was isolated from a denitrification reactor treating a closed-circuit seawater aquarium. It can sustain growth under anoxic conditions by reducing nitrate (NO3-) to nitrite (NO2-), which accumulates in the medium. These physiological traits are attributed to gene clusters that encode two dissimilatory nitrate reductases (NarGHJI). M. nitratireducenticrescens strain JAM1 also contains gene clusters encoding two putative nitric oxide reductase (NO) reductases and one putative nitrous oxide (N2O) reductase, suggesting that NO and N2O can be reduced by strain JAM1. In this study, we show that strain JAM1 can reduce NO to N2O and N2O to N2 and can sustain growth under anoxic conditions by reducing N2O as the sole electron acceptor. Although strain JAM1 lacks a gene encoding a dissimilatory copper-(NirK) or cytochrome cd1-type (NirS) NO2- reductase, NO3--amended strain JAM1 cultures produce N2O, representing up to 6% of the N-input. NO2- was shown to be the key intermediate of this production process. In NO3-amended cultures, we analyzed denitrification genes in succession of net N2O-production and -consumption phases at the gene expression level. These phases were found to correlate with changes in the expression levels of the NO reductase gene cnorB1 and nnrS, which indicated NO production in the cultures.\n\nImportanceBy showing that all the three denitrification reductases are active, this demonstrates that Methylophaga nitratireducenticrescens JAM1 is one of many bacteria species that maintain genes associated primarily with denitrification, but not necessarily related to the maintenance of the entire pathway. The reason to maintain such incomplete pathway could be related to the specific role of strain JAM1 in the denitrifying biofilm of a denitrification reactor from which it originates. The small production of N2O via NO in strain JAM1 did not involve Nar contrary to what was demonstrated in Escherichia coli. M. nitratireducenticrescens JAM1 is the only reported Methylophaga species that has the capacity to grow under anoxic conditions by using NO3- and N2O as sole electron acceptors for its growth. It is also one of a few marine methylotrophs that is studied at the physiological and genetic levels in relation to its capacity to perform denitrifying activities.

microbiology↗