Bacteria of the genus Dehalococcoides play an important role in the reductive dechlorination of chlorinated ethenes. A systemslevel approach was taken in this study to examine the global transcriptomic and proteomic responses of exponentially growing cells of Dehalococcoides ethenogenes strain 195 to fixed nitrogen limitation (FNL), as dechlorination activity and cell yield both decrease during FNL. As expected, the nitrogen-fixing (nif) genes were differentially upregulated in the transcriptome and proteome of strain 195 during FNL. Aside from the nif operon, a putative methylglyoxal synthase-encoding gene (DET1576), the product of which is predicted to catalyze the formation of the toxic electrophile methylglyoxal and is implicated in the uncoupling of anabolism from catabolism in bacteria, was strongly upregulated in the transcriptome and could potentially play a role in the observed growth inhibition during FNL. Carbon catabolism genes were generally downregulated in response to FNL, and a number of transporters were differentially regulated in response to nitrogen limitation, with some playing apparent roles in nitrogen acquisition, while others were associated with general stress responses. A number of genes related to the functions of nucleotide synthesis, replication, transcription, translation, and posttranslational modifications were also differentially expressed. One gene coding for a putative reductive dehalogenase (DET1545) and a number of genes coding for oxidoreductases, which have implications in energy generation and redox reactions, were also differentially regulated. Interestingly, most of the genes within the multiple integrated elements were not differentially expressed. Overall, this study elucidates the molecular responses of strain 195 to FNL and identifies differentially expressed genes that are potential biomarkers to evaluate environmental cellular nitrogen status.
Bacteria of the genus Dehalococcoides play a key role in the bioremediation of the carcinogenic and toxic chlorinated ethenes because of their ability to reductively dechlorinate tetrachloroethene (PCE) and its daughter products to the innocuous product ethene (9,10,29,36,49). While bacteria from diverse genera can dechlorinate PCE and trichloroethene (TCE) to isomers of dichloroethene (DCE), to date, dechlorination beyond DCE to vinyl chloride (VC) and ethene has been observed only for strains of Dehalococcoides (47). Through transcriptional and protein analyses, the reductive dehalogenase (RDase)-and hydrogenase-encoding genes for energy generation in Dehalococcoides isolates and enrichments have been studied extensively (6,16,19,23,27,28,34,36,39,40,55). However, other aspects of the physiology of Dehalococcoides are not as well understood.Currently, the genomes of four Dehalococcoides strains have been sequenced, annotated, and published (20,30,45). A nitrogenase-encoding operon (nif) for the reduction of atmospheric dinitrogen to ammonia is present in the genome annotation of Dehalococcoides ethenogenes strain 195 (45) ...