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Volume 71, Issue 2

Mutants of the α-Ketoglutarate Dehydrogenase Gene of k12 Free

Abstract

SUMMARY

Of 21 mutants of k12 five were shown to be mutants by their sensitivity to suppressors. In addition to lacking activity for the overall Α-ketoglutarate dehydrogenase complex and the Α-ketoglutarate dehydrogenase component (E1), the mutants had little or no dihydrolipoyl trans-succinylase (E2) and 20 to 30% less dihydrolipoyl dehydrogenase (E3) than the parental strain. In common with non-suppressible mutants they also had reduced amounts of succinylCoA synthetase and succinate dehydrogenase. Fine structure mapping by Pi-transduction showed that some of the sites were located in the gene. It is concluded that expression of the region is polarized from to , i.e. synthesis of the E1 component precedes that of the E2 component. Double mutants with lesions in the pyruvate and Α-ketoglutarate dehydrogenase genes () were constructed but their E3 activity was never less than 30% of that of the parental strain. The results are consistent with the existence of separate genes for the dihydrolipoyl dehydrogenase component of the pyruvate and Α-ketoglutarate dehydrogenase complexes, but other possibilities could not be ruled out. Further studies on the positions of the four closely-linked tricarboxylic acid cycle genes specifying citrate synthase (), succinate dehydrogenase () and two components of the Α-ketoglutarate dehydrogenase complex ( and ) indicate the relative order:

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© Society for General Microbiology, 1962
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1972-07-01
2024-04-25
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Amber Mutants of the α-Ketoglutarate Dehydrogenase Gene of Escherichia coli k12
Microbiology 71, 207 (1972); https://doi.org/10.1099/00221287-71-2-207
/content/journal/micro/10.1099/00221287-71-2-207
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