Pyoverdine-mediated iron uptake in Pseudomonas aeruginosa: the Tat system is required for PvdN but not for FpvA transport.
Under iron-limiting conditions, Pseudomonas aeruginosa PAO1 secretes a fluorescent siderophore called pyoverdine (Pvd). After chelating iron, this ferric siderophore is transported back into the cells via the outer membrane receptor FpvA. The Pvd-dependent iron uptake pathway requires several essential genes involved in both the synthesis of Pvd and the uptake of ferric Pvd inside the cell. A previous study describing the global phenotype of a tat-deficient P. aeruginosa strain showed that the defect in Pvd-mediated iron uptake was due to the Tat-dependent export of proteins involved in Pvd biogenesis and ferric Pvd uptake (U. Ochsner, A. Snyder, A. I. Vasil, and M. L. Vasil, Proc. Natl. Acad. Sci. USA 99:8312-8317, 2002). Using biochemical and biophysical tools, we showed that despite its predicted Tat signal sequence, FpvA is correctly located in the outer membrane of a tat mutant and is fully functional for all steps of the iron uptake process (ferric Pvd uptake and recycling of Pvd on FpvA after iron release). However, in the tat mutant, no Pvd was produced. This suggested that a key element in the Pvd biogenesis pathway must be exported to the periplasm by the Tat pathway. We located PvdN, a still unknown but essential component in Pvd biogenesis, at the periplasmic side of the cytoplasmic membrane and showed that its export is Tat dependent. Our results further support the idea that a critical step of the Pvd biogenesis pathway involving PvdN occurs at the periplasmic side of the cytoplasmic membrane.
Voulhoux
Romé
R
IBSM-LISM, CNRS, Marseille Cedex, France. voulhoux@ibsm.cnrs-mrs.fr
Filloux
Alain
A
Schalk
Isabelle J
IJ
D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism; D001426:Bacterial Proteins / Q000378:metabolism / Q000502:physiology*; D003593:Cytoplasm / Q000378:metabolism; D029968:Escherichia coli Proteins; D007501:Iron / Q000378:metabolism*; D026901:Membrane Transport Proteins / Q000502:physiology*; D008566:Membranes / Q000378:metabolism; D009842:Oligopeptides / Q000378:metabolism*; D010453:Peptide Synthases / Q000378:metabolism; D011550:Pseudomonas aeruginosa / Q000378:metabolism*
D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; C082326:FpvA protein, Pseudomonas aeruginosa; D026901:Membrane Transport Proteins; D009842:Oligopeptides; C408585:twin-arginine translocase complex, E coli; C042453:pyoverdin; D007501:Iron; D010453:Peptide Synthases