Bacteria of the genus Shewanella are known for their versatile electron-accepting capacities, which allow them to couple the decomposition of organic matter to the reduction of the various terminal electron acceptors that they encounter in their stratified environments. Owing to their diverse metabolic capabilities, shewanellae are important for carbon cycling and have considerable potential for the remediation of contaminated environments and use in microbial fuel cells. Systems-level analysis of the model species Shewanella oneidensis MR-1 and other members of this genus has provided new insights into the signal-transduction proteins, regulators, and metabolic and respiratory subsystems that govern the remarkable versatility of the shewanellae.
Horizontal gene transfer contributes to the evolution of bacterial species. Mobile genetic elements play an important role in horizontal gene transfer, and characterization of the regulation of these elements should provide insight into conditions that influence bacterial evolution. We characterized a mobile genetic element, ICEBs1, in the Gram-positive bacterium Bacillus subtilis and found that it is a functional integrative and conjugative element (ICE) capable of transferring to Bacillus and Listeria species. We identified two conditions that promote ICEBs1 transfer: conditions that induce the global DNA damage response and crowding by potential recipients that lack ICEBs1. Transfer of ICEBs1 into cells that already contain the element is inhibited by an intercellular signaling peptide encoded by ICEBs1. The dual regulation of ICEBs1 allows for passive propagation in the host cell until either the potential mating partners lacking ICEBs1 are present or the host cell is in distress.conjugation ͉ horizontal gene transfer ͉ quorum sensing ͉ peptide signaling ͉ DNA microarrays
Clostridium difficile disease has recently increased to
become a dominant nosocomial pathogen in North America and Europe, although
little is known about what has driven this emergence. Here we show two epidemic
ribotypes (RT027 and RT078) have acquired unique mechanisms to metabolize low
concentrations of the disaccharide trehalose. RT027 strains contain a single
point mutation in the trehalose repressor that increases this ribotype’s
sensitivity to trehalose by >500 fold. Furthermore, dietary trehalose
increases virulence of a RT027 strain in a mouse model of infection. RT078
strains acquired a cluster of four genes involved in trehalose metabolism,
including a PTS permease that is both necessary and sufficient for growth on low
concentrations of trehalose. We propose that the implementation of trehalose as
a food additive into the human diet, shortly before the emergence of these two
epidemic lineages, helped select for their emergence and contributed to
hypervirulence.
SummaryICEBs1 is a mobile genetic element found in the chromosome of Bacillus subtilis. Excision and transfer of ICEBs1 is regulated by the global DNA damage response and intercellular peptide signalling. We identified and characterized a repressor, ImmR (formerly YdcN), encoded by ICEBs1. ImmR represses transcription of genes required for excision and transfer, and both activates and represses its own transcription. ImmR regulates transcription within ICEBs1 by binding to several sites in the region of DNA that contains promoters for both immR and xis (encoding excisionase). In addition, we found that ImmR confers immunity from acquisition of additional copies of ICEBs1. ImmR-mediated regulation serves to keep a single copy of ICEBs1 stably maintained in the absence of induction, allows a rapid response to inducing signals, and helps limit acquisition of additional copies of ICEBs1.
SummaryICEBs1 is an integrative and conjugative element (conjugative transposon) integrated into trnS-leu2 in Bacillus subtilis. In response to DNA damage or high concentrations of potential mating partners, ICEBs1 can excise and transfer to various recipients, including other species. We found that excision of ICEBs1 occurs by site-specific recombination within 60 bp direct repeats that mark the junctions between ICEBs1 and chromosomal DNA. Excision required two ICEBs1 genes, int (integrase, ydcL), predicted to encode a tyrosine recombinase similar to that of phage lambda, and xis (excisionase, sacV). Ectopic expression of xis was sufficient to induce excision of ICEBs1, indicating that regulation of xis transcription by DNA damage and peptide signalling normally controls excision. Int, but not Xis, was needed for sitespecific integration. We found that in the absence of the primary bacterial attachment site (attB) in trnSleu2, ICEBs1 integrated in secondary attachment sites that are similar to a 17 bp sequence in attB. In the absence of int, ICEBs1 could recombine into the chromosome by RecA-dependent homologous recombination, provided ICEBs1 contained a region of sequence identity to a chromosomal locus.
We sought to identify the fungi that colonize healthy GI tracts and that have a sustained influence on the diverse functions of the gut microbiome. Instead, we found that all fungi in the stool of healthy volunteers could be explained by their presence in oral and dietary sources and that our results, together with those from other analyses, support the model that there is little or no gastrointestinal colonization by fungi. This may be due to Westernization, primate evolution, fungal ecology, and/or the strong defenses of a healthy immune system. Importantly, fungal colonization of the GI tract may often be indicative of disease. As fungi can cause serious infections in immunocompromised individuals and are found at increased abundance in multiple disorders of the GI tract, understanding normal fungal colonization is essential for proper treatment and prevention of fungal pathogenesis.
BackgroundContinuous-flow culture models are one tool for studying complex interactions between members of human fecal microbiotas because they allow studies to be completed during an extended period of time under conditions where pH, nutrient availability, and washout of waste products and dead cells can be controlled. Because many of the existing well-validated continuous-flow models are large and complex, we were interested in developing a simpler continuous-flow system that would allow microbial community dynamics to be examined in higher throughput while still maintaining complex microbial communities. To this end, we developed minibioreactor arrays (MBRAs), small volume bioreactors (15 ml) that allow simultaneous cultivation of up to 48 microbial communities in a single anaerobic chamber.ResultsWe used MBRA to characterize the microbial community dynamics of replicate reactors inoculated from three different human fecal donors and reactors seeded with feces pooled from these three donors. We found that MBRA could be used to efficiently cultivate complex microbial communities that were a subset of the initial fecal inoculum (15–25 % of fecal OTUs initially observed). After an initial acclimation period of approximately 1 week, communities in each reactor stabilized and exhibited day-to-day variation similar to that observed in stable mouse fecal communities. Replicate reactors were predominately populated by shared core microbial communities; variation between replicate reactors was primarily driven by shifts in abundance of shared operational taxonomic units (OTUs). Consistent with differences between fecal donors, MBRA communities present in reactors seeded with different fecal samples had distinct composition and structure.ConclusionsFrom these analyses, we conclude that MBRAs can be used to cultivate communities that recapitulate key features of human fecal communities and are a useful tool to facilitate higher-throughput studies of the dynamics of these communities.Electronic supplementary materialThe online version of this article (doi:10.1186/s40168-015-0106-5) contains supplementary material, which is available to authorized users.
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