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PMID:4590465

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Contents

Citation

Guardiola, J, De Felice, M, Klopotowski, T and Iaccarino, M (1974) Mutations affecting the different transport systems for isoleucine, leucine, and valine in Escherichia coli K-12. J. Bacteriol. 117:393-405

Abstract

Uptake of isoleucine, leucine, and valine in Escherichia coli K-12 is due to several transport processes for which kinetic evidence has been reported elsewhere. A very-high-affinity transport process, a high-affinity transport process, and three different low-affinity transport processes were described. In this paper the existence of these transport processes is confirmed by the isolation and preliminary characterization of mutants altered in one or more of them. The very-high-affinity transport process is missing either in strains carrying the brnR6(am) mutation or in strains carrying the brn-8 mutation. This appears to be a pleiotropic effect since other transport systems are also missing. Mutant analysis shows that more than one transport system with high affinity is present. One of them, high-affinity 1, which needs the activity of a protein produced by the brnQ gene, transports isoleucine, leucine, and valine and is unaffected by threonine. The other, high-affinity 2, which needs the activity of a protein produced by the brnS gene, transports isoleucine, leucine, and valine; this uptake is inhibited by threonine which probably is a substrate. Another protein, produced by the brnR gene, is required for uptake through both high-affinity 1 and high-affinity 2 transport systems. The two systems therefore appear to work in parallel, brnR being a branching point. The brnQ gene is located close to phoA at 9.5 min on the chromosome of E. coli, the brnR gene is located close to lac at 9.0 min, and the brnS gene is close to pdxA at 1 min. A mutant lacking the low-affinity transport system for isoleucine was isolated from a strain in which the high-affinity system was missing because of a brnR mutation. This strain also required isoleucine for growth because of an ilvA mutation. The mutant lacking the low-affinity transport system was unable to grow on isoleucine but could grow on glycylisoleucine. This mutant had lost the low-affinity transport for isoleucine, whereas those for leucine and valine were unaffected. A pleiotropic consequence of this mutation (brn-8) was a complete absence of the very-high-affinity transport system due either to the alteration of a common gene product or to any kind of secondary interference which inhibits it. Mutants altered in isoleucine-leucine-valine transport were isolated by taking advantage of the inhibition that valine exerts on the K-12 strain of E. coli. Mutants resistant both to valine inhibition (Val(r)) and to glycylvaline inhibition are regulatory mutants. Val(r) mutants that are sensitive to glycylvaline inhibition are transport mutants. When the very-high-affinity transport process is repressed (for example by methionine) the frequency of transport mutants among Val(r) mutants is higher, and it is even higher if the high-affinity transport process is partially inhibited by leucine.

Links

PubMed PMC285526

Keywords

Alanine/pharmacology; Amino Acids/pharmacology; Bacterial Proteins/metabolism; Biological Transport, Active; Chromosome Mapping; Escherichia coli/growth & development; Escherichia coli/metabolism; Genes; Isoleucine/metabolism; Leucine/metabolism; Methionine/pharmacology; Mutagens; Mutation; Nitrosoguanidines; Threonine/pharmacology; Transduction, Genetic; Ultraviolet Rays; Valine/metabolism

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Annotations

Gene product Qualifier GO ID GO term name Evidence Code with/from Aspect Notes Status

brnQ:Gene_Product(s)

GO:0015820

leucine transport

IMP: Inferred from Mutant Phenotype

P

Seeded from EcoCyc (v14.0)

complete

brnQ:Gene_Product(s)

GO:0015803

branched-chain aliphatic amino acid transport

IMP: Inferred from Mutant Phenotype

P

complete

brnQ:Gene_Product(s)

GO:0015829

valine transport

IMP: Inferred from Mutant Phenotype

P

complete

brnQ:Gene_Product(s)

GO:0015818

isoleucine transport

IMP: Inferred from Mutant Phenotype

P

complete

brnQ:Gene_Product(s)

GO:0015820

leucine transport

IMP: Inferred from Mutant Phenotype

P

inhibited by threonine

complete

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