Mutations affecting the regulation of γ-aminobutyrate utilization inEscherichia coli K-12

Abstract

Four genes,gabCPDT, are involved in the utilization of γ-aminobutyrate (GABA) byEscherichia coli K-12. Thegab gene cluster maps nearrecA andsrl, at 57.5 min.gabP, gabD andgabT specify the synthesis of GABA transport carrier, succinic semialdehyde dehydrogenase (SSDH), and glutamate-succinic semialdehyde transaminase (GSST), respectively;gabC controls the synthesis of all three proteins. GABA-nonutilizing mutants carrying deletions insrl extended into thegab cluster have been isolated. The mutants completely lost the capacity for GABA transport, while preserving full activity of GSST and SSDH, suggesting thatgabC is not a promoter-operator locus or a gene coding for an activator protein. A mutation ingabD (M-16) that abolished SSDH activity had the following additional properties: It exerted a bipolar effect on the neighboring genes, greatly reducing the activities of GSST and SSDH; the polar effect ongabP but not ongabT was fully suppressed by the knownrho mutation suA78; at least three classes of GABA-utilizing revertants of M-16 were obtained: (i) revertants with allgab activities restored to the parental levels; (ii) revertants with SSDH activity still missing, but with the other activities fully repaired; (iii) revertants with no SSDH activity, with GSST partly recovered, but with transport fully repaired. It is suggested that thegab cluster is transcribed bidirectionally from a promoter in thegabD region and that the mutation in strain M-16 may be due to DNA insertion in that region.