Identification of livG, a membrane-associated component of the branched-chain amino acid transport in Escherichia coli.
Branched-chain amino acids are transported into Escherichia coli by two osmotic shock-sensitive systems (leucine-isoleucine-valine and leucine-specific transport systems). These high-affinity systems consist of separate periplasmic binding protein components and at least three common membrane-bound components. In this study, one of the membrane-bound components, livG, was identified. A toxic analog of leucine, azaleucine, was used to isolate a large number of azaleucine-resistant mutants which were defective in branched-chain amino acid transport. Genetic complementation studies established that two classes of transport mutants with similar phenotypes, livH and livG, were obtained which were defective in one of the membrane-associated transport components. Since the previously cloned plasmid, pOX1, genetically complemented both livH and livG mutants, we were able to verify the physical location of the livG gene on this plasmid. Recombinant plasmids which carried different portions of the pOX1 plasmid were constructed and subjected to complementation analysis. These results established that livG was located downstream from livH with about 1 kilobase of DNA in between. The expression of these plasmids was studied in minicells; these studies indicate that livG appears to be membrane bound and to have a molecular weight of 22,000. These results establish that livG is a membrane-associated component of the branched-chain amino acid transport system in E. coli.
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Mayo
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Anderson
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Oxender
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D000596:Amino Acids / Q000378:metabolism*; D001426:Bacterial Proteins; D001693:Biological Transport, Active; D002352:Carrier Proteins / Q000235:genetics*; D002462:Cell Membrane / Q000378:metabolism; D004262:DNA Restriction Enzymes; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D029968:Escherichia coli Proteins; D005020:Ethyl Methanesulfonate / Q000494:pharmacology; D005796:Genes; D005798:Genes, Bacterial; D005816:Genetic Complementation Test; D005838:Genotype; D009154:Mutation; D010957:Plasmids; D013045:Species Specificity
D000596:Amino Acids; D001426:Bacterial Proteins; D002352:Carrier Proteins; D029968:Escherichia coli Proteins; C488923:LivJ protein, E coli; C028332:leucine-isoleucine-valine binding protein, bacteria; D005020:Ethyl Methanesulfonate; D004262:DNA Restriction Enzymes