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Integrated Analysis of Protein Complexes and Regulatory Networks Involved in Anaerobic Energy Metabolism of Shewanella Oneidensis MR-1

Description: Anaerobic Nitrate Reduction. Nitrate is an extensive co-contaminant at some DOE sites making metal and radionuclide reduction problematic. Hence, we sought to better understand the nitrate reduction pathway and its control in S. oneidensis MR-1. It is not known whether the nitrate reduction is by denitrification or dissimilatory nitrate reduction into ammonium (DNRA). By both physiological and genetic evidence, we proved that DNRA is the nitrate reduction pathway in this organism. Using the complete genome sequence of S. oneidensis MR-1, we identified a gene encoding a periplasmic nitrate reductase based on its 72% sequence identity with the napA gene in E. coli. Anaerobic growth of MR-1 on nitrate was abolished in a site directed napA mutant, indicating that NapA is the only nitrate reductase present. The anaerobic expression of napA and nrfA, a homolog of the cytochrome b552 nitrite reductase in E. coli, increased with increasing nitrate concentration until a plateau was reached at 3 mM KNO3. This indicates that these genes are not repressed by increasing concentrations of nitrate. The reduction of nitrate can generate intermediates that can be toxic to the microorganism. To determine the genetic response of MR-1 to high concentrations of nitrate, DNA microarrays were used to obtain a complete gene expression profile of MR-1 at low (1 mM) versus high (40 mM) nitrate concentrations. Genes encoding transporters and efflux pumps were up-regulated, perhaps as a mechanism to export toxic compounds. In addition, the gene expression profile of MR-1, grown anaerobically with nitrate as the only electron acceptor, suggested that this dissimilatory pathway contributes to N assimilation. Hence the nitrate reduction pathway could serve a dual purpose. The role of EtrA, a homolog of Fnr (global anaerobic regulator in E. coli) was examined using an etrA deletion mutant we constructed, S. oneidensis EtrA7-1.
Date: June 1, 2005
Creator: Tiedje, James M.
Partner: UNT Libraries Government Documents Department

Noncompetitive microbial diversity patterns in soils: their causes and implications for bioremediation

Description: This funding provided support for over nine years of research on the structure and function of microbial communities in subsurface environments. The overarching goal during these years was to understand the impact of mixed contaminants, particularly heavy metals like uranium, on the structure and function of microbial communities. In addition we sought to identify microbial populations that were actively involved in the reduction of metals because these species of bacteria hold the potential for immobilizing soluble metals moving in subsurface water. Bacterial mediated biochemical reduction of metals like uranium, technetium and chromium, greatly reduces their mobility through complexation and precipitation. Hence, by taking advantage of natural metabolic capabilities of subsurface microbial populations it is possible to bioremediate contaminated subsurface environments with a cost-effective in situ approach. Towards this end we have i.) identified bacterial populations that have thrived under the adverse conditions at the contaminated FRC site, ii.) phylogenetically identified populations that respond to imposed remediation conditions at the FRC, iii.) used metagenomics to begin a reconstruction of the metabolic web in a contaminated subsurface zone, iv.) investigated the metal reducing attributes of a Gram-positive spore forming rod also capable of dechlorination.
Date: July 5, 2007
Creator: Tiedje, James M.; Zhou, Jizhong; Palumbo, Anthony; Ostrom, Nathaniel & Marsh, Terence L.
Partner: UNT Libraries Government Documents Department

Integrated Genome-Based Studies of Shewanella Ecophysiology

Description: The aim of the work reported is to study Shewanella population genomics, and to understand the evolution, ecophysiology, and speciation of Shewanella. The tasks supporting this aim are: to study genetic and ecophysiological bases defining the core and diversification of Shewanella species; to determine gene content patterns along redox gradients; and to Investigate the evolutionary processes, patterns and mechanisms of Shewanella.
Date: January 8, 2014
Creator: Tiedje, James M.; Konstantinidis, Kostas & Worden, Mark
Partner: UNT Libraries Government Documents Department

Metagenomic Insights into Evolution of a Heavy Metal-Contaminated Groundwater Microbial Community

Description: Understanding adaptation of biological communities to environmental change is a central issue in ecology and evolution. Metagenomic analysis of a stressed groundwater microbial community reveals that prolonged exposure to high concentrations of heavy metals, nitric acid and organic solvents (~;;50 years) have resulted in a massive decrease in species and allelic diversity as well as a significant loss of metabolic diversity. Although the surviving microbial community possesses all metabolic pathways necessary for survival and growth in such an extreme environment, its structure is very simple, primarily composed of clonal denitrifying ?- and ?-proteobacterial populations. The resulting community is over-abundant in key genes conferring resistance to specific stresses including nitrate, heavy metals and acetone. Evolutionary analysis indicates that lateral gene transfer could be a key mechanism in rapidly responding and adapting to environmental contamination. The results presented in this study have important implications in understanding, assessing and predicting the impacts of human-induced activities on microbial communities ranging from human health to agriculture to environmental management, and their responses to environmental changes.
Date: February 15, 2010
Creator: Hemme, Christopher L.; Deng, Ye; Gentry, Terry J.; Fields, Matthew W.; Wu, Liyou; Barua, Soumitra et al.
Partner: UNT Libraries Government Documents Department