Suppression of the btuB451 mutation by mutations in the tonB gene suggests a direct interaction between TonB and TonB-dependent receptor proteins in the outer membrane of Escherichia coli

Abstract

In cells of Escherichia coli, the function of the tonB gene is needed for energy-dependent transport processes mediated by the outer-membrane receptors for iron siderophore complexes and vitamin B12. The btuB451 mutation has the same effect on vitamin B12 transport as does a tonB mutation. When a btuB451 strain carried a plasmid with the intact tonB gene, partial revertant strains were isolated which had acquired the ability to grow on 5 nM vitamin B12. This suppression activity was associated with the plasmid, suggesting that a mutation within the tonB gene on the plasmid allowed the mutant BtuB receptor to function in the transport of the vitamin. The nucleotide sequence of the entire tonB gene of ten independently isolated suppressing plasmids was determined. Only a single nucleotide change had occurred in each of the cases. The same codon was always affected resulting in the conversion of glutamine-165 to a leucine in seven of the ten isolates and to a lysine in the other three. The phenotype of strains carrying both types of altered tonB genes showed the retention of their function for other TonB-dependent processes in addition to their suppressor properties with respect to the btuB451 mutation. The fact that mutations suppressing the btuB451 mutation occurred in the tonB gene suggests that there is a direct interaction between TonB and TonB-dependent receptors in the outer membrane of E. coli.