mBio最新文献

The composition of the gut microbiome is determined by a complex interplay of diet, host genetics, microbe-microbe interactions, abiotic factors, and stochasticity. Previous studies have demonstrated the importance of host genetics in community assembly of the Caenorhabditis elegans gut microbiome and identified a central role for DBL-1/BMP immune signaling in determining the abundance of gut Enterobacteriaceae. However, the effects of DBL-1 signaling on gut bacteria were found to depend on its activation in extra-intestinal tissues, highlighting a gap in our understanding of the proximal factors that determine microbiome composition. In the present study, we used RNA-seq gene expression analysis of wildtype, dbl-1 and sma-3 mutants, and dbl-1 over-expressors to identify candidate DBL-1/BMP targets that may mediate the pathway's effects on gut commensals. Bacterial colonization experiments in mutants, or following RNAi-mediated knock-down of candidate genes specifically in the intestine, demonstrated their local contribution to intestinal control of Enterobacteriaceae abundance. Furthermore, epistasis analysis suggested that these contributions were downstream of the DBL-1 pathway, together suggesting that examined candidates were intestinal effectors and mediators of DBL-1 signaling, contributing to the shaping of gut microbiome composition.IMPORTANCECompared to the roles of diet, environmental availability, or lifestyle in determining gut microbiome composition, that of genetic factors is the least understood and often underestimated. The identification of intestinal effectors of distinct molecular functions that control enteric bacteria offers a glimpse into the genetic logic of microbiome control as well as a list of targets for future exploration of this logic.

Fitness optimization in a dynamic environment requires bacteria to adapt their proteome in a tightly regulated manner by altering protein production and/or degradation. Here, we investigate proteome adaptation in Lactococcus cremoris following a sudden nutrient upshift (e.g., nutrients that allow faster growth) and focus especially on the fate of redundant proteins after the shift. Protein turnover analysis demonstrated that L. cremoris cultures shifted from galactose to glucose, immediately accelerate growth and initiate proteome-wide adjustment toward glucose-optimized composition. Redundant proteins were predominantly adjusted by lowering (or stopping) protein production combined with dilution by growth. However, pyruvate formate lyase activator (PflA) was actively degraded, which appears correlated to reduced 4Fe-4S cofactor availability. Active PflA removal induces the shutdown of galactose-associated mixed acid fermentation to accelerate the switch toward glucose-associated homolactic fermentation. Our work deciphers molecular adjustments upon environmental change that drive physiological adaptation, including growth rate and central energy metabolism.IMPORTANCEBacteria adapt to their environment by adjusting their molecular makeup, in particular their proteome, which ensures fitness optimization under the newly encountered environmental condition. We present a detailed analysis of proteome adaptation kinetics in Lactococcus cremoris following its acute transition from galactose to glucose media, as an example of a sudden nutrient quality upshift. Analysis of the replacement times of individual proteins after the nutrient upshift established that the entire proteome is instantly adjusting to the new condition, which coincides with immediate growth rate acceleration and metabolic adaptation. The latter is driven by the active removal of the pyruvate formate lyase activator protein that is pivotal in controlling pyruvate dissipation in L. cremoris. Our work exemplifies the amazing rate of molecular adaptation in bacteria that underlies physiological adjustments, including growth rate and carbon metabolism. This mechanistic study contributes to our understanding of adaptation in L. cremoris during the dynamic conditions it encounters during (industrial) fermentation, even though environmental transitions in these processes are mostly more gradual than the acute shift studied here.

Climate change is predicted to increase the spread of mosquito-borne viruses, but genetic mechanisms underlying the influence of environmental variation on the ability of insect vectors to transmit human pathogens is unknown. In response to a changing climate, mosquitoes will experience longer periods of drought. An important physiological response to dry environments is the protection against dehydration, here defined as desiccation tolerance. While temperature is known to impact interactions between mosquito and virus, the role of dehydration remains unknown. We identified two genetically diverse lines of the mosquito Aedes aegypti, a major arbovirus vector, with marked differences in desiccation tolerance. To determine the genetic response to dehydration between these contrasting lines, we compared gene expression profiles between desiccant- and non-desiccant-treated individuals in both the desiccation-tolerant and -susceptible lines by RNAseq. Gene expression analysis demonstrated that several genes are differentially expressed in response to desiccation stress between desiccation-tolerant and -susceptible lines. The most highly expressed transcript under desiccation stress in the desiccation-susceptible line encodes a peritrophin protein, Ae-Aper50. Peritrophins play a crucial role in peritrophic matrix formation in the mosquito midgut after a bloodmeal. Gene silencing of Ae-Aper50 by RNAi demonstrated that expression of Ae-Aper50 is required for survival of the desiccation-susceptible line under desiccation stress, but not for the desiccation-tolerant line. Moreover, the knockdown of Ae-Aper50 resulted in higher Zika virus (ZIKV) infection rates in the desiccation-tolerant line and increased ZIKV viral replication in the desiccation susceptible line, and higher chikungunya virus (CHIKV) infection rates in the desiccation-tolerant line. Altogether, these results provide a link between protection against desiccation and midgut infection, which has important implications in predicting how climate change will impact mosquito-borne viruses.

Importance: Climate change will have profound impacts on the burden of viruses transmitted by mosquitoes. While we know how changes in temperature impact mosquito physiology and dynamics of viral replication within the mosquito, there is a complete lack of knowledge in how low humidity, or drought tolerance, will impact interactions between mosquitoes and arboviruses. Understanding how drought tolerance will alter mosquito infection with arboviruses is critical in predicting and preventing the impact that climate change will have on mosquito-borne viruses. This work demonstrates a functional link between dehydration tolerance and midgut infection. This knowledge significantly enhances our understanding of how the predicted increase in droughts could impact the dynamics of mosquito-borne viruses.

Mycobacterium tuberculosis (Mtb) exhibits an impressive ability to adapt to rapidly changing environments, despite its genome's apparent stability. Recently, phase variation through indel formation in homopolymeric tracts (HT) has emerged as a potentially important mechanism promoting adaptation in Mtb. This study examines the impact of common phase variants associated with the ESX-1 type VII secretion system, focusing on a highly variable HT upstream of the ESX-1 regulatory factor, espR. By engineering this frequently observed indel into an isogenic background, we demonstrate that a single nucleotide insertion in the espR 5'UTR causes post-transcriptional upregulation of EspR protein abundance and corresponding alterations in the EspR regulon. Consequently, this mutation increases the expression of ESX-1 components in the espACD operon and enhances ESX-1 substrate secretion. We find that this indel specifically increases isoniazid resistance without impacting the effectiveness of other drugs tested. Furthermore, we show that two distinct observed HT indels that regulate either espR translation or espACD transcription increase bacterial fitness in a mouse infection model. The presence of multiple ESX-1-associated HTs provides a mechanism to combinatorially tune protein secretion, drug sensitivity, and host-pathogen interactions. More broadly, these findings support emerging data that Mtb utilizes HT-mediated phase variation to direct genetic variation to certain sites across the genome in order to adapt to changing pressures.

Importance: Mycobacterium tuberculosis (Mtb) is responsible for more deaths worldwide than any other single infectious agent. Understanding how this pathogen adapts to the varied environmental pressures imposed by host immunity and antibiotics has important implications for the design of more effective therapies. In this work, we show that the genome of Mtb contains multiple contingency loci that control the activity of the ESX-1 secretion system, which is critical for interactions with the host. These loci consist of homopolymeric DNA tracts in gene regulatory regions that are subject to high-frequency reversible variation and act to tune the activity of ESX-1. We find that variation at these sites increases the fitness of Mtb in the presence of antibiotic and/or during infection. These findings indicate that Mtb has the ability to diversify its genome in specific sites to create subpopulations of cells that are preadapted to new conditions.

Bacteriophages (phages) are being investigated as potential biocontrol agents for the suppression of bacterial diseases in cultivated crops. Jumbo bacteriophages, which possess genomic DNA larger than 200 kbp, generally have a broader host range than other phages and therefore would be useful as biocontrol agents against a wide range of bacterial strains. Thus, the characterization of novel jumbo phages specific for agricultural pathogens would be of importance for the development of phage biocontrol strategies. Herein, we demonstrate that phage S13 requires Burkholderia glumae flagella for its attachment and infection and that loss of B. glumae flagella prevents S13 cellular lysis. As flagella is a known virulence factor, loss of flagella results in a surviving population of B. glumae with reduced virulence. Further experimentation demonstrates that phage S13 can protect rice plants from B. glumae-sponsored destruction in a rice seedling model of infection.IMPORTANCEBacterial plant pathogens threaten many major food crops and inflict large agricultural losses worldwide. B. glumae is a bacterial plant pathogen that causes diseases such as rot, wilt, and blight in several food major crops including rice, tomato, hot pepper, and eggplant. B. glumae infects rice during all developmental stages, causing diseases such as rice seedling rot and bacterial panicle blight (BPB). The B. glumae incidence of rice plant infection is predicted to increase with warming global temperatures, and several different control strategies targeting B. glumae are being explored. These include chemical and antibiotic soil amendment, microbiome manipulation, and the use of partially resistant rice cultivars. However, despite rice growth amelioration, the treatment options for B. glumae plant infections remain limited to cultural practices. Alternatively, phage biocontrol represents a promising new method for eliminating B. glumae from crop soils and improving rice yields.

Bats are reservoirs for multiple viruses, some of which are known to cause global disease outbreaks. Virus spillovers from bats have been implicated in zoonotic transmission. Some bat species can tolerate viral infections, such as infections with coronaviruses and paramyxoviruses, better than humans and with less clinical consequences. Bat species are speculated to have evolved alongside these viral pathogens, and adaptations within the bat immune system are considered to be associated with viral tolerance. Inflammation and cell death in response to zoonotic virus infections prime human immunopathology. Unlike humans, bats have evolved adaptations to mitigate virus infection-induced inflammation. Inflammatory cell death pathways such as necroptosis and pyroptosis are associated with immunopathology during virus infections, but their regulation in bats remains understudied. This review focuses on the regulation of inflammation and cell death pathways in bats. We also provide a perspective on the possible contribution of cell death-regulating proteins, such as caspases and gasdermins, in modulating tissue damage and inflammation in bats. Understanding the role of these adaptations in bat immune responses can provide valuable insights for managing future disease outbreaks, addressing human disease severity, and improving pandemic preparedness.