A phosphatidylserine-binding site in the cytosolic fragment of Clostridium sordellii lethal toxin facilitates glucosylation of membrane-bound Rac and is required for cytotoxicity.
Large clostridial toxins glucosylate some small G proteins on a threonine residue, thereby preventing their interactions with effector molecules and regulators. We show that the glucosyltransferase domain of lethal toxin from Clostridium sordellii (LT(cyt); amino acids 1-546), which is released into the cytosol during cell infection, binds preferentially to liposomes containing phosphatidylserine as compared with other anionic lipids. The binding of LT(cyt) to phosphatidylserine increases by two orders of magnitude the rate of glucosylation of liposome-bound geranyl-geranylated Rac-GDP. Limited proteolysis and deletion studies show that the binding site for phosphatidylserine lies within the first 18 N-terminal residues of LT(cyt). Deletion of these residues abolishes the effect of phosphatidylserine on the activity of LT(cyt) on liposome-bound geranyl-geranylated Rac-GDP and prevents the morphological effects induced by LT(cyt) microinjection into various cells, but it does not affect the intrinsic activity of LT(cyt) on non-geranyl-geranylated Rac-GDP in solution. We conclude that the avidity of LT(cyt) for phosphatidylserine facilitates its targeting to the cytosolic leaflet of cell membranes and, notably, the plasma membrane, where this anionic lipid is abundant and where several targets of lethal toxin reside.
Mesmin
Bruno
B
CNRS, Institut de Pharmacologie Moléculaire et Cellulaire, 660 Route des Lucioles, 06560 Valbonne, France.
Robbe
Karine
K
Geny
Blandine
B
Luton
Frédéric
F
Brandolin
Gérard
G
Popoff
Michel R
MR
Antonny
Bruno
B
D001427:Bacterial Toxins / Q000378:metabolism*; D047989:Paraclostridium sordellii / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D006031:Glycosylation / Q000187:drug effects; D008081:Liposomes / Q000378:metabolism; D010718:Phosphatidylserines / Q000378:metabolism* / Q000494:pharmacology; D020744:rac GTP-Binding Proteins / Q000378:metabolism*
D001427:Bacterial Toxins; D008081:Liposomes; D010718:Phosphatidylserines; C072292:lethal toxin LT, Clostridium sordellii; D020744:rac GTP-Binding Proteins