Identification of the vibriobactin receptor of Vibrio cholerae.
Vibrio cholerae produces the novel phenolate siderophore vibriobactin and several outer membrane proteins in response to iron starvation. To determine whether any of these iron-regulated outer membrane proteins serves as the receptor for vibriobactin, the classical V. cholerae strain 0395 was mutagenized by using TnphoA, and iron-regulated fusions were analyzed for vibriobactin transport. One mutant, MBG14, was unable to bind or utilize exogenous vibriobactin and did not grow in low-iron medium. However, synthesis of the siderophore and transport of other iron complexes, including ferrichrome, hemin, and ferric citrate, were unaffected in MBG14. Analysis of membrane proteins by sodium dodecyl sulfate-polyacrylamide gel electrophoresis demonstrated the loss from the mutant of a 74-kDa iron-regulated outer membrane protein present in the parental strain when grown in iron-limiting conditions. This protein partitioned into the detergent phase during Triton X-114 extraction, suggesting that it is a hydrophobic membrane protein. DNA sequences encoding the gene into which TnphoA had inserted, designated viuA (vibriobactin uptake), restored the wild-type phenotype to the mutant; the complemented mutant expressed the 74-kDa outer membrane protein under iron-limiting conditions and possessed normal vibriobactin binding and uptake. These data indicate that the 74-kDa outer membrane protein of V. cholerae serves as the vibriobactin receptor.
Stoebner
J A
JA
Department of Microbiology, University of Texas, Austin 78712-1095.
Butterton
J R
JR
Calderwood
S B
SB
Payne
S M
SM
D001425:Bacterial Outer Membrane Proteins / Q000235:genetics*; D002396:Catechols / Q000378:metabolism*; D004252:DNA Mutational Analysis; D004251:DNA Transposable Elements; D015964:Gene Expression Regulation, Bacterial; D007501:Iron / Q000378:metabolism; D016254:Mutagenesis, Insertional; D010080:Oxazoles; D011956:Receptors, Cell Surface / Q000235:genetics*; D014734:Vibrio cholerae / Q000235:genetics*
D001425:Bacterial Outer Membrane Proteins; D002396:Catechols; D004251:DNA Transposable Elements; D010080:Oxazoles; D011956:Receptors, Cell Surface; C035902:siderophore receptors; C040227:vibriobactin; D007501:Iron