A leucine-rich motif targets Pseudomonas aeruginosa ExoS within mammalian cells.
Type III cytotoxins contribute to the ability of bacterial pathogens to subvert the host innate immune system. ExoS (453 amino acids) is a bifunctional type III cytotoxin produced by Pseudomonas aeruginosa. Residues 96 to 232 comprise a Rho GTPase activating protein domain, while residues 233 to 453 comprise a 14-3-3-dependent ADP-ribosyltransferase domain. An N-terminal domain (termed the membrane localization domain [MLD]) targets ExoS to the Golgi-endoplasmic reticulum (Golgi-ER) of mammalian cells. This study identifies an amino acid motif that is responsible for the membrane binding properties of the MLD. Deletion mapping showed that the MLD included a symmetrical leucine-rich motif within residues 51 to 77 of ExoS. The terminal dileucines and internal leucines and an isoleucine within the MLD, but not charged or other hydrophobic residues, targeted a reporter protein to the Golgi-ER region of HeLa cells. Mutations of the leucines within the MLD did not affect type III secretion or translocation into HeLa cells but limited the ability of ExoS to ADP-ribosylate Ras GTPases. Mutations of charged residues within the MLD did not affect type III secretion, delivery into HeLa cells, or the ability of ExoS to ADP-ribosylate Ras GTPases. The organization of the leucines within the MLD of ExoS is different from that of previously described leucine-rich motifs but is present in several other bacterial proteins. This implies a role for intracellular targeting in the efficient targeting of mammalian cells by type III cytotoxins.
Zhang
Yue
Y
Medical College of Wisconsin, Microbiology and Molecular Genetics, 8701 Watertown Plank Rd., Milwaukee, WI 53226, USA.
Barbieri
Joseph T
JT
D036002:ADP Ribose Transferases / Q000032:analysis / Q000737:chemistry* / Q000378:metabolism*; D020727:ADP-Ribosylation Factors / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D020816:Amino Acid Motifs / Q000235:genetics; D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D001427:Bacterial Toxins / Q000032:analysis / Q000737:chemistry* / Q000378:metabolism*; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism / Q000382:microbiology; D003603:Cytotoxins / Q000032:analysis / Q000737:chemistry* / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D007930:Leucine / Q000235:genetics / Q000502:physiology*; D008969:Molecular Sequence Data; D009154:Mutation; D021381:Protein Transport; D011550:Pseudomonas aeruginosa / Q000378:metabolism / Q000472:pathogenicity*; D017384:Sequence Deletion; D018631:ras Proteins / Q000378:metabolism
D001427:Bacterial Toxins; D003603:Cytotoxins; D036002:ADP Ribose Transferases; C444820:exoenzyme S; D020727:ADP-Ribosylation Factors; D018631:ras Proteins; D007930:Leucine