Browsing by Author "Hamilton, Trinity L."

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Aerobic and Anaerobic Thiosulfate Oxidation by a Cold-Adapted, Subglacial Chemoautotroph
(2015-12) Harrold, Zoe R.; Skidmore, Mark L.; Hamilton, Trinity L.; Desch, Elizabeth; Kirina, Amada; van Gelder, Will; Glover, Kevin; Roden, Eric E.; Boyd, Eric S.
Geochemical data indicate that protons released during pyrite (FeS2) oxidation are important drivers of mineral weathering in oxic and anoxic zones of many aquatic environments including those beneath glaciers. Oxidation of FeS2 under oxic, circumneutral conditions proceeds through the metastable intermediate thiosulfate (S2O32-), which represents an electron donor capable of supporting microbial metabolism. Subglacial meltwaters sampled from Robertson Glacier (RG), Canada over a seasonal melt cycle reveal concentrations of S2O32- that are typically below detection despite the presence of available pyrite and several orders of magnitude higher concentrations of the FeS2 oxidation product sulfate (SO42-). Here we report the physiological and genomic characterization of the chemolithoautotrophic facultative anaerobe Thiobacillus sp. RG5 isolated from the subglacial environment at RG. The RG5 genome encodes pathways for the complete oxidation of S2O32-, CO2 fixation, and aerobic and anaerobic respiration with nitrite or nitrate. Growth experiments indicate that the energy required to synthesize a cell under oxygen or nitrate reducing conditions with S2O32- as electron donor was lower at 5.1 °C than 14.4 °C, indicating that this organism is cold-adapted. RG sediment-associated soxB transcripts, which encode a component of the S2O32--oxidizing complex, were closely affiliated to soxB from RG5. Collectively, these results suggest an active sulfur cycle in the subglacial environment at RG mediated in part by populations closely affiliated with RG5. Microbial consumption of S2O32- by RG5-like populations may accelerate abiotic FeS2 oxidation thereby enhancing mineral weathering in the subglacial environment.
Chemolithotrophic primary production in a subglacial ecosystem
(2014-10) Boyd, Eric S.; Hamilton, Trinity L.; Havig, Jeff R.; Skidmore, Mark L.; Shock, Everett L.
Glacial comminution of bedrock generates fresh mineral surfaces capable of sustaining chemotrophic microbial communities under the dark conditions that pervade subglacial habitats. Geochemical and isotopic evidence suggests that pyrite oxidation is a dominant weathering process generating protons that drive mineral dissolution in many subglacial systems. Here, we provide evidence correlating pyrite oxidation with chemosynthetic primary productivity and carbonate dissolution in subglacial sediments sampled from Robertson Glacier (RG), Alberta, Canada. Quantification and sequencing of ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) transcripts suggest that populations closely affiliated with Sideroxydans lithotrophicus, an iron sulfide-oxidizing autotrophic bacterium, are abundant constituents of microbial communities at RG. Microcosm experiments indicate sulfate production during biological assimilation of radiolabeled bicarbonate. Geochemical analyses of subglacial meltwater indicate that increases in sulfate levels are associated with increased calcite and dolomite dissolution. Collectively, these data suggest a role for biological pyrite oxidation in driving primary productivity and mineral dissolution in a subglacial environment and provide the first rate estimate for bicarbonate assimilation in these ecosystems. Evidence for lithotrophic primary production in this contemporary subglacial environment provides a plausible mechanism to explain how subglacial communities could be sustained in near-isolation from the atmosphere during glacial-interglacial cycles.
Draft genome sequence of a sulfide-oxidizing, autotrophic filamentous anoxygenic phototrophic bacterium, Chloroflexus sp. strain MS-G (Chloroflexi)
(American Society for Microbiology, 2014-09) Thiel, Vera; Hamilton, Trinity L.; Tomsho, Lynn P.; Burhans, Richard; Gay, Scott E.; Schuster, Stephan C.; Ward, David M.; Bryant, Donald A.
The draft genome sequence of the thermophilic filamentous anoxygenic phototrophic bacterium Chloroflexus sp. strain MS-G (Chloroflexi), isolated from Mushroom Spring (Yellowstone National Park, WY, USA) was sequenced and comprises 4,784,183 bp in 251 contigs. The draft genome is predicted to encode 4,059 protein coding genes, 49 tRNA encoding genes, and 3 rRNA operons.
Effect of salinity on mercury methylating benthic microbes and their activities in Great Salt Lake, Utah
(2017-03) Boyd, Eric S.; Yu, Ri-Qing; Barkay, Tamar; Hamilton, Trinity L.; Baxter, Bonnie K.; Naftz, David L.; Marvin-DiPasquale, Mark
Surface water and biota from Great Salt Lake (GSL) contain some of the highest documented concentrations of total mercury (THg) and methylmercury (MeHg) in the United States. In order to identify potential biological sources of MeHg and controls on its production in this ecosystem, THg and MeHg concentrations, rates of Hg(II)-methylation and MeHg degradation, and abundances and compositions of archaeal and bacterial 16 rRNA gene transcripts were determined in sediment along a salinity gradient in GSL. Rates of Hg(II)-methylation were inversely correlated with salinity and were at or below the limits of detection in sediment sampled from areas with hypersaline surface water. The highest rates of Hg(II)-methylation were measured in sediment with low porewater salinity, suggesting that benthic microbial communities inhabiting less saline environments are supplying the majority of MeHg in the GSL ecosystem. The abundance of 16S rRNA gene transcripts affiliated with the sulfate reducer Desulfobacterium sp. was positively correlated with MeHg concentrations and Hg(II)-methylation rates in sediment, indicating a potential role for this taxon in Hg(II)-methylation in low salinity areas of GSL. Reactive inorganic Hg(II) (a proxy used for Hg(II) available for methylation) and MeHg concentrations were inversely correlated with salinity. Thus, constraints imposed by salinity on Hg(II)-methylating populations and the availability of Hg(II) for methylation are inferred to result in higher MeHg production potentials in lower salinity environments. Benthic microbial MeHg degradation was also most active in lower salinity environments. Collectively, these results suggest an important role for sediment anoxia and microbial sulfate reducers in the production of MeHg in low salinity GSL sub-habitats and may indicate a role for salinity in constraining Hg(II)-methylation and MeHg degradation activities by influencing the availability of Hg(II) for methylation.
[FeFe]-Hydrogenase Abundance and Diversity along a Vertical Redox Gradient in Great Salt Lake, USA
(2014-12) Boyd, Eric S.; Hamilton, Trinity L.; Swanson, Kevin D.; Howells, Alta E.; Meuser, Jonathan E.; Posewitz, Matthew C.; Peters, John W.
The use of [FeFe]-hydrogenase enzymes for the biotechnological production of H2 or other reduced products has been limited by their sensitivity to oxygen (O2). Here, we apply a PCR-directed approach to determine the distribution, abundance, and diversity of hydA gene fragments along co-varying salinity and O2 gradients in a vertical water column of Great Salt Lake (GSL), UT. The distribution of hydA was constrained to water column transects that had high salt and relatively low O2 concentrations. Recovered HydA deduced amino acid sequences were enriched in hydrophilic amino acids relative to HydA from less saline environments. In addition, they harbored interesting variations in the amino acid environment of the complex H-cluster metalloenzyme active site and putative gas transfer channels that may be important for both H2 transfer and O2 susceptibility. A phylogenetic framework was created to infer the accessory cluster composition and quaternary structure of recovered HydA protein sequences based on phylogenetic relationships and the gene contexts of known complete HydA sequences. Numerous recovered HydA are predicted to harbor multiple N- and C-terminal accessory iron-sulfur cluster binding domains and are likely to exist as multisubunit complexes. This study indicates an important role for [FeFe]-hydrogenases in the functioning of the GSL ecosystem and provides new target genes and variants for use in identifying O2 tolerant enzymes for biotechnological applications.
Geobiological feedbacks and the evolution of thermoacidophiles
(2017-10) Colman, Daniel R.; Poudel, Saroj; Hamilton, Trinity L.; Havig, Jeff R.; Selensky, Matthew J.; Shock, Everett L.; Boyd, Eric S.
Oxygen-dependent microbial oxidation of sulfur compounds leads to the acidification of natural waters. How acidophiles and their acidic habitats evolved, however, is largely unknown. Using 16S rRNA gene abundance and composition data from 72 hot springs in Yellowstone National Park, Wyoming, we show that hyperacidic (pH<3.0) hydrothermal ecosystems are dominated by a limited number of archaeal lineages with an inferred ability to respire O2. Phylogenomic analyses of 584 existing archaeal genomes revealed that hyperacidophiles evolved independently multiple times within the Archaea, each coincident with the emergence of the ability to respire O2, and that these events likely occurred in the recent evolutionary past. Comparative genomic analyses indicated that archaeal thermoacidophiles from independent lineages are enriched in similar protein-coding genes, consistent with convergent evolution aided by horizontal gene transfer. Because the generation of acidic environments and their successful habitation characteristically require O2, these results suggest that thermoacidophilic Archaea and the acidity of their habitats co-evolved after the evolution of oxygenic photosynthesis. Moreover, it is likely that dissolved O2 concentrations in thermal waters likely did not reach levels capable of sustaining aerobic thermoacidophiles and their acidifying activity until ~0.8Ga, when present day atmospheric levels were reached, a time period that is supported by our estimation of divergence times for archaeal thermoacidophilic clades.
Toward Better Genetically Encoded Sensors of Membrane Potential
(2016-03) Urschel, Matthew R.; Hamilton, Trinity L.; Roden, Eric E.; Boyd, Eric S.
Facultative autotrophs are abundant components of communities inhabiting geothermal springs. However, the influence of uptake kinetics and energetics on preference for substrates is not well understood in this group of organisms. Here, we report the isolation of a facultatively autotrophic crenarchaeote, strain CP80, from Cinder Pool (CP, 88.7°C, pH 4.0), Yellowstone National Park. The 16S rRNA gene sequence from CP80 is 98.8% identical to that from Thermoproteus uzonensis and is identical to the most abundant sequence identified in CP sediments. Strain CP80 reduces elemental sulfur (S8°) and demonstrates hydrogen (H2)-dependent autotrophic growth. H2-dependent autotrophic activity is suppressed by amendment with formate at a concentration in the range of 20–40 μM, similar to the affinity constant determined for formate utilization. Synthesis of a cell during growth with low concentrations of formate required 0.5 μJ compared to 2.5 μJ during autotrophic growth with H2. These results, coupled to data indicating greater C assimilation efficiency when grown with formate as compared to carbon dioxide, are consistent with preferential use of formate for energetic reasons. Collectively, these results provide new insights into the kinetic and energetic factors that influence the physiology and ecology of facultative autotrophs in high-temperature acidic environments.