Project description:The ubiquitous heterotrophic marine bacterium, Rugeria pomeroyi, was experimentally cultured under both environmentally realistic carbon conditions and with a tracer-level addition of 13C-labeled leucine. Bacterial protein biosynthesis was tracked through exponential and stationary growth phases. This combination of methods allowed for observation of real-time bacterial protein production of an environmentally relevant marine bacterium under low-carbon conditions to understand metabolic priorities during different growth phases.
Project description:The heterotrophic marine bacterium, Ruegeria pomeroyi, was experimentally cultured under environmentally realistic carbon conditions and with a tracer-level addition of 13C-labeled leucine to track bacterial protein biosynthesis through growth phases. A combination of methods allowed observation of real-time bacterial protein production to understand metabolic priorities through the different growth phases. Over 2000 proteins were identified in each experimental culture from exponential and stationary growth phases. Within two hours of the 13C-labeled leucine addition, R. pomeroyi significantly assimilated the newly encountered substrate into new proteins. This dataset provides a fundamental baseline for understanding growth phase differences in molecular physiology of a cosmopolitan marine bacterium.
Project description:Microbial alteration and remineralization of organic matter in the ocean play essential roles in the carbon cycle. The impact of individual compounds as either growth substrates or infochemicals is still poorly constrained in models, particularly for organisms with relatively versatile genomes. Here, we use metabolomics techniques to characterize the response of a heterotrophic marine bacterium, Ruegeria pomeroyi DSS-3, to growth on the abundant algal metabolite dimethylsulfoniopropionate (DMSP). DMSP is a valuable source of reduced carbon and sulfur as well as a chemo-attractant. It is present at higher concentrations during phytoplankton blooms. When cultivated on DMSP, R. pomeroyi synthesized a quorum sensing molecule, N-(3-oxotetradecanoyl)-L-homoserine lactone, which was not synthesized at the same levels during growth on propionate, another carbon substrate. More broadly, we observed differential production of intra- and extracellular metabolites with implications for nitrogen assimilation and microbial cross-feeding. This multi-faceted response suggests that R. pomeroyi uses a two-component regulatory system dependent on the presence of DMSP and high cell densities to activate a suite of metabolic changes that may shape how it accesses nutrients and impacts the surrounding microbial consortium.
Project description:The goal of this project was to identify bacterial transporters responsible for uptake of environmentally relevant marine metabolites. We used the model marine heterotrophic bacterium Ruegeria pomeroyi DSS-3, for which an arrayed library of single gene knockout mutants has been generated by selecting isolated from a barcoded transposon mutant library (BasSeq). Knockout mutants of putative transporters were grown on minimal medium with a single substrate as sole carbon source. Mutant defect was assessed by comparing the substrate drawdown of isolated mutants to drawdown by a pooled mutant library (BarSeq), a proxy for wildtype fitness.
Project description:When the genome of Ruegeria pomeroyi DSS-3 was published in 2004, it represented the first sequence from a heterotrophic marine bacterium. Over the last ten years, the strain has become a valuable model for understanding the cycling of sulfur and carbon in the ocean. To ensure that this genome remains useful, we have updated 69 genes to incorporate functional annotations based on new experimental data, and improved the identification of 120 protein-coding regions based on proteomic and transcriptomic data. We review the progress made in understanding the biology of R. pomeroyi DSS-3 and list the changes made to the genome.
Project description:Marine phototroph and heterotroph interactions are vital in maintaining the nutrient balance in the oceans as essential nutrients need to be rapidly cycled before sinking to aphotic layers. The aim of this study was to highlight the molecular mechanisms that drive these interactions. For this, we generated a detailed exoproteomic time-course analysis of a 100-day co-culture between the model marine picocyanobacterium Synechococcus sp. WH7803 and the Roseobacter strain Ruegeria pomeroyi DSS-3, both in nutrient-enriched and natural oligotrophic seawater. The proteomic data showed a transition between the initial growth phase and stable-state phase that, in the case of the heterotroph, was caused by a switch in motility attributed to organic matter availability. The phototroph adapted to seawater oligotrophy by reducing its selective leakiness, increasing the acquisition of essential nutrients and secreting conserved proteins of unknown function. We also report a surprisingly high abundance of extracellular superoxide dismutase produced by Synechococcus and a dynamic secretion of potential hydrolytic enzyme candidates used by the heterotroph to cleave organic groups and hydrolase polymeric organic matter produced by the cyanobacterium. The time course dataset we present here will become a reference for understanding the molecular processes underpinning marine phototroph-heterotroph interactions.