Activation of Akt/protein kinase B in epithelial cells by the Salmonella typhimurium effector sigD.
The serine-threonine kinase Akt is a protooncogene involved in the regulation of cell proliferation and survival. Activation of Akt is initiated by binding to the phospholipid products of phosphoinositide 3-kinase at the inner leaflet of the plasma membranes followed by phosphorylation at Ser(473) and Thr(308). We have found that Akt is activated by Salmonella enterica serovar Typhimurium in epithelial cells. A bacterial effector protein, SigD, which is translocated into host cells via the specialized type III secretion system, is essential for Akt activation. In HeLa cells, wild type S. typhimurium induced translocation of Akt to membrane ruffles and phosphorylation at residues Thr(308) and Ser(473) and increased kinase activity. In contrast, infection with a SigD deletion mutant did not induce phosphorylation or activity although Akt was translocated to membrane ruffles. Complementation of the SigD deletion strain with a mutant containing a single Cys to Ser mutation (C462S), did not restore the Akt activation phenotype. This residue has previously been shown to be essential for inositol phosphatase activity of the SigD homologue, SopB. Our data indicate a novel mechanism of Akt activation in which the endogenous cellular pathway does not convert membrane-associated Akt into its active form. SigD is also the first bacterial effector to be identified as an activator of Akt.
Steele-Mortimer
O
O
Biotechnology Laboratory and Department of Medicine, University of British Columbia, Vancouver, British Columbia V6T 1Z3, Canada. osteelem@cellbio.wustl.edu
Knodler
L A
LA
Marcus
S L
SL
Scheid
M P
MP
Goh
B
B
Pfeifer
C G
CG
Duronio
V
V
Finlay
B B
BB
D001483:Base Sequence; D017931:DNA Primers; D004789:Enzyme Activation / Q000502:physiology; D004847:Epithelial Cells / Q000201:enzymology*; D005408:Flagellin / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D021381:Protein Transport; D011518:Proto-Oncogene Proteins; D051057:Proto-Oncogene Proteins c-akt; D012486:Salmonella typhimurium / Q000378:metabolism*
D017931:DNA Primers; D005408:Flagellin; D019869:Phosphatidylinositol 3-Kinases; D017346:Protein Serine-Threonine Kinases; D011518:Proto-Oncogene Proteins; D051057:Proto-Oncogene Proteins c-akt; C416532:flaB flagellin; C494918:AKT1 protein, human