ADP-ribosylation and functional effects of Pseudomonas exoenzyme S on cellular RalA.
Exoenzyme S (ExoS) is a bifunctional virulence factor directly translocated into eukaryotic cells by the type III secretory process of Pseudomonas aeruginosa. Bacterial translocation of ExoS into epithelial cells is associated with diverse effects on cell function, including inhibition of growth, alterations in cell morphology, and effects on adherence processes. Preferred substrates of the ADP-ribosyltransferase (ADPRT) portion of ExoS include low molecular weight G-proteins (LMWG-proteins) in the Ras family. In examining the ADP-ribosylation and functional effects of ExoS on RalA, ExoS was found to ADP-ribosylate endogenous RalA and recombinant RalADeltaCAAX at multiple sites, with Arg52 identified as the preferred site of ADP-ribosylation. The binding of RalA to the Ral binding domain (RBD) of its downstream effector, RalBP1, was inhibited by bacterially translocated ExoS, indicating an effect of ExoS on cellular RalA function. In vitro analyses confirmed that ADP-ribosylation of RalA directly interfered with its ability to bind to the RBD of RalBP1. The studies support the fact that RalA is a cellular substrate of bacterially translocated ExoS and that ADP-ribosylation by ExoS affects RalA interaction with its downstream effector, RalBP1.
Department of Pathology and Laboratory Medicine, Medical University of South Carolina, 165 Ashley Avenue, Suite 309, Charleston, South Carolina 29425, USA.
Jennifer E
JE
Fraylick
Matthew J
MJ
Riese
Timothy S
TS
Vincent
Joseph T
JT
Barbieri
Joan C
JC
Olson
D036002:ADP Ribose Transferases / Q000378:metabolism*; D000246:Adenosine Diphosphate Ribose / Q000378:metabolism*; D001427:Bacterial Toxins; D001483:Base Sequence; D017931:DNA Primers; D015180:Electrophoresis, Gel, Two-Dimensional; D020558:GTP Phosphohydrolases / Q000378:metabolism*; D011549:Pseudomonas / Q000201:enzymology*
D001427:Bacterial Toxins; D017931:DNA Primers; D000246:Adenosine Diphosphate Ribose; D036002:ADP Ribose Transferases; C444820:exoenzyme S; D020558:GTP Phosphohydrolases