Mutational analysis of a bifunctional ferrisiderophore receptor and signal-transducing protein from Pseudomonas aeruginosa.
The FpvA protein of Pseudomonas aeruginosa strain PAO1 mediates uptake of a siderophore, ferripyoverdine. It is also a component of a signal transduction pathway that controls production of an exotoxin, a protease, pyoverdine, and FpvA itself. The purpose of the research described here was to dissect these different functions of FpvA. Signaling involves an N-terminal domain of FpvA, and it was shown that this domain is probably located in the periplasm, as expected. Short peptides were inserted at 36 sites within FpvA by linker insertion mutagenesis. The effects of these mutations on the presence of FpvA in the outer membrane, on FpvA-mediated uptake of ferripyoverdine, and on pyoverdine synthesis and gene expression were determined. Five of the mutations resulted in the absence of FpvA from the outer membrane of the bacteria. All of the remaining mutations eliminated either the transport or signaling function of FpvA and most affected both functions. Three mutations prevented transport of ferripyoverdine but had no effect on the signal transduction pathway showing that transport of ferripyoverdine is not required for the trans-membrane signaling process. Conversely, eight mutations affected pyoverdine-mediated signaling but had no effect on transport of ferripyoverdine. These data show that insertions throughout FpvA resulted in loss of function and that signaling and transport are separate and discrete functions of FpvA.
James
H Ellen
HE
Department of Biochemistry, University of Otago, PO Box 56, Dunedin, New Zealand.
Beare
Paul A
PA
Martin
Lois W
LW
Lamont
Iain L
IL
D001425:Bacterial Outer Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism; D004252:DNA Mutational Analysis; D005290:Ferric Compounds / Q000378:metabolism; D009842:Oligopeptides / Q000096:biosynthesis / Q000378:metabolism; D019897:Periplasm / Q000378:metabolism; D017434:Protein Structure, Tertiary; D011550:Pseudomonas aeruginosa / Q000378:metabolism*; D017262:Siderophores / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D015398:Signal Transduction
D001425:Bacterial Outer Membrane Proteins; D005290:Ferric Compounds; C082326:FpvA protein, Pseudomonas aeruginosa; D009842:Oligopeptides; D017262:Siderophores; C042453:pyoverdin