Investigating cellular responses to mutations in the glutathione and thioredoxin pathways of Escherichia coli

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Investigating cellular responses to mutations in the glutathione and thioredoxin pathways of Escherichia coli

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Title: Investigating cellular responses to mutations in the glutathione and thioredoxin pathways of Escherichia coli
Author: Chrysostomou, Constantine
Abstract: Inhibition of disulfide bond formation in Escherichia coli implicates an intricate collaboration of proteins which comprise the glutathione and thioredoxin reducing pathways. Bioengineers have successfully engineered E. coli possessing mutated reducing pathways that promote, rather than inhibit, disulfide bond formation in the cytoplasm. The transcriptome of six such mutant E. coli strains have been characterized using Microarray technology. We find that all mutant strains, exhibit a unique response to oxidative stress, not observed in wild type. Statistical analyses revealed the expression of more than 200 genes that are affected by mutations within the reducing pathways. Significantly up-regulated biological processes include cysteine biosynthesis, histidine biosynthesis, NADH Dehydrogenase I biosynthesis, sugar catabolic processes, and activation of stress responses . The second part of this work describes the construction of an E. coli strain that promotes the complete conversion of glutathione into its seemingly dormant derivative, glutathionylspermidine. This engineered strain can be used in assays designed to evaluate the effectiveness of glutathionylspermidine as a substitute for glutathione and, hopefully, allude to its true metabolic function.
Subject: Microarray Glutathione Thioredoxin Oxidative stress Glutathionylspermidine
URI: http://hdl.handle.net/2152/ETD-UT-2009-12-706
Date: 2009-12

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