Abstract
Mutations in two chromosomal genes of Escherichia coli, cpxA and cpxB, produced a temperature-sensitive growth defect that was remedied specifically by the addition of isoleucine and valine to the minimal medium. This auxotrophy was manifested only when the medium contained exogenous leucine, suggesting that mutant cells fail to elaborate active acetohydroxy acid synthase, isozyme I. In the presence of leucine, this enzyme was required to catalyze the first reaction common to the biosynthesis of isoleucine and valine. Measurements of enzyme activity in crude extracts showed that mutant cells were seven- to eightfold deficient in active isozyme I when the cells were grown in the presence of leucine. When grown in the absence of leucine, mutant cells contained more acetohydroxy acid synthase activity. We attribute this activity to isozyme III, the product of the ilvHI genes, which are derepressed in the absence of exogenous leucine. The cpxA and cpxB mutations appear to affect the production of active isozyme I, rather than its activity, since (i) neither the cpxA nor the cpxB gene mapped near the structural gene for isozyme I (ilvB), (ii) the growth of mutant cells shifted from the permissive (34 degrees C) to the nonpermissive (41 degrees C) temperature did not immediately cease, but declined gradually over a period corresponding to several normal generation times, and (iii) the enzyme from mutant cells grown at 34 degrees C was as stable at 41 degrees C as the enzyme from cpx+ cells.
MeSH Terms
Acetolactate Synthase/genetics
Escherichia coli/genetics,metabolism
Genes
Isoenzymes/genetics
Isoleucine/biosynthesis
Leucine
Mutation
Oxo-Acid-Lyases/genetics
Temperature
Valine/biosynthesis
Chemicals
Isoenzymes
Isoleucine
Acetolactate Synthase
Oxo-Acid-Lyases
Leucine
Valine
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
McEwen J
Silverman P
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15 references, click to expand
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