Project description:Copper is a highly reactive, toxic metal; consequently, transport of this metal within the cell is tightly regulated. Intriguingly, the actinobacterium Kineococcus radiotolerans has been shown to not only accumulate soluble copper to high levels within the cytoplasm, but the phenotype also correlated with enhanced cell growth during chronic exposure to ionizing radiation. This study offers a first glimpse into the physiological and proteomic responses of K. radiotolerans to copper at increasing concentration and distinct growth phases. Aerobic growth rates and biomass yields were similar over a range of Cu(II) concentrations (0-1.5 mM) in complex medium. Copper uptake coincided with active cell growth and intracellular accumulation was positively correlated with Cu(II) concentration in the growth medium (R(2)=0.7). Approximately 40% of protein coding ORFs on the K. radiotolerans genome were differentially expressed in response to the copper treatments imposed. Copper accumulation coincided with increased abundance of proteins involved in oxidative stress and defense, DNA stabilization and repair, and protein turnover. Interestingly, the specific activity of superoxide dismutase was repressed by low to moderate concentrations of copper during exponential growth, and activity was unresponsive to perturbation with paraquot. The biochemical response pathways invoked by sub-lethal copper concentrations are exceptionally complex; though integral cellular functions are preserved, in part, through the coordination of defense enzymes, chaperones, antioxidants and protective osmolytes that likely help maintain cellular redox. This study extends our understanding of the ecology and physiology of this unique actinobacterium that could potentially inspire new biotechnologies in metal recovery and sequestration, and environmental restoration.
Project description:In this study, approximately 36 and 29 million raw reads of two samples, namely radiation treated strain and its untreated control, are acquired from the sequencing platform. And 143 genes are screened out with the differential expression (DE) analysis.
Project description:Kineococcus radiotolerans SRS30216 was isolated from a high-level radioactive environment at the Savannah River Site (SRS) and exhibits gamma-radiation resistance approaching that of Deinococcus radiodurans. The genome was sequenced by the U.S. Department of Energy's Joint Genome Institute which suggested the existence of three replicons, a 4.76 Mb linear chromosome, a 0.18 Mb linear plasmid, and a 12.92 Kb circular plasmid. Southern hybridization confirmed that the chromosome is linear. The K. radiotolerans genome sequence was examined to learn about the physiology of the organism with regard to ionizing radiation resistance, the potential for bioremediation of nuclear waste, and the dimorphic life cycle. K. radiotolerans may have a unique genetic toolbox for radiation protection as it lacks many of the genes known to confer radiation resistance in D. radiodurans. Additionally, genes involved in the detoxification of reactive oxygen species and the excision repair pathway are overrepresented. K. radiotolerans appears to lack degradation pathways for pervasive soil and groundwater pollutants. However, it can respire on two organic acids found in SRS high-level nuclear waste, formate and oxalate, which promote the survival of cells during prolonged periods of starvation. The dimorphic life cycle involves the production of motile zoospores. The flagellar biosynthesis genes are located on a motility island, though its regulation could not be fully discerned. These results highlight the remarkable ability of K radiotolerans to withstand environmental extremes and suggest that in situ bioremediation of organic complexants from high level radioactive waste may be feasible.
Project description:While a part of the native gut microflora, the Gram-positive bacterium Enterococcus faecalis can lead to serious infections elsewhere in the body. The draft genome of E. faecalis strain ATCC BAA-2128, isolated from piglet feces, was examined. This draft genome consists of 42 contigs, 12 of which exhibit homology to annotated plasmids.
Project description:The physiological and transcriptomic response of the metal resistant bacterium Cupriavidus metallidurans strain CH34 in response to stable (non-radioactive) strontium ions (Sr) was investigated. C. metallidurans CH34 could survive and proliferate in the presence of relatively high concentrations of SrCl2 (D10 is 70mM, MIC is 120 mM). Precipitation of Sr as strontium carbonate was observed in the culture during aerobic growth of CH34 in the presence of 60 mM SrCl2. To identify the cellular mechanisms involved in the bioprecipitation process, gene expression in the cells was analyzed after short-time (30 min) exposure to low (5 mM) and high (60 mM) concentrations of SrCl2. The transcription of the gene clusters annotated as hmyFCBA and czcCBADRS, coding for ion efflux pumps, was significantly induced following exposure to Sr, and not with Ca. There were also significant changes is the transcription of the genes encoding TctCBA proteins involved in citrate uptake and two hypothetical porin coding genes following exposure Sr and Ca. These results highlight a specific molecular response of bacterium Cupriavidus metallidurans CH34 to Sr, including the identification of putative Sr specific efflux pumps, and thus the potential of this bacterium to distinguish Sr from Ca. These findings will help to better understand natural Sr (and Ca) microbial weathering or mineralization processes in the environment, and could be of interest for bioremediation or bioprocessing of (radioactive) Sr-containing water, soil or waste. Two-condition experiments. Comparing samples after induction with metals (Sr, Ca) versus non-induced samples. Biological triplicate. Each array contains 3 technical replicates.