1887

Abstract

produces, under conditions of iron limitation, a high-affinity siderophore, pyoverdine (PVD), which is recognized at the level of the outer membrane by a specific TonB-dependent receptor, FpvA. So far, for , three different PVDs, differing in their peptide chain, have been described (types I–III), but only the FpvA receptor for type I is known. Two PVD-producing strains, one type II and one type III, were mutagenized by a mini-Tn transposon. In each case, one mutant unable to grow in the presence of the strong iron chelator ethylenediaminedihydroxyphenylacetic acid (EDDHA) and the cognate PVD was selected. The first mutant, which had an insertion in the gene, upstream of , was unable to take up type II PVD and showed resistance to pyocin S3, which is known to use type II FpvA as receptor. The second mutant was unable to take up type III PVD and had the transposon insertion in . Cosmid libraries of the respective type II and type III PVD wild-type strains were constructed and screened for clones restoring the capacity to grow in the presence of PVD. From the respective complementing genomic fragments, type II and type III sequences were determined. When , type II and type III restored PVD production, uptake, growth in the presence of EDDHA and, in the case of type II , pyocin S3 sensitivity. Complementation of mutants obtained by allelic exchange was achieved by the presence of cognate . All three receptors posses an N-terminal extension of about 70 amino acids, similar to FecA of , but only FpvAI has a TAT export sequence at its N-terminal end.

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2003-04-01
2024-04-18
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