YopH dephosphorylates Cas and Fyn-binding protein in macrophages.
The tyrosine phosphatase YopH is an essential virulence effector of pathogenic Yersinia spp. YopH, which is translocated from extracellularly located bacteria into interacting target cells, blocks phagocytosis by professional phagocytes. We show here that immunoprecipitation of YopH from lysates of J774 cells infected with Y. pseudotuberculosis expressing an inactive form of YopH resulted in co-precipitation of certain phosphotyrosine proteins. The association between the inactive YopH and phosphotyrosine proteins in the 120 kDa range was rapid and could be detected after 2 min of infection. The proteins were identified as the docking proteins Cas and Fyn-binding protein (FYB). Upon infection of J774 cells with Y. pseudotuberculosis lacking YopH expression both of these proteins became tyrosine phosphorylated. Moreover, this infection caused recruitment of Cas to peripheral focal complexes, and FYB was relocalized to areas surrounding these structures. Both Cas and FYB became dephosphorylated upon infection with Y. pseudotuberculosis expressing active YopH, and this was associated with disruption of focal complexes. With regard to the previous identification of Cas and focal complexes as targets of YopH in HeLa cells, the present study supports an important role for these targets in a general mechanism of bacterial uptake.
Hamid
N
N
Department of Cell and Molecular Biology, University of Umeâ, Umeâ, S-901 87, Sweden.
Gustavsson
A
A
Andersson
K
K
McGee
K
K
Persson
C
C
Rudd
C E
CE
Fällman
M
M
D001425:Bacterial Outer Membrane Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D002352:Carrier Proteins / Q000302:isolation & purification / Q000378:metabolism*; D051177:Catenins; D015815:Cell Adhesion Molecules / Q000302:isolation & purification / Q000378:metabolism*; D002460:Cell Line; D005455:Fluorescent Antibody Technique; D008264:Macrophages; D010750:Phosphoproteins / Q000302:isolation & purification / Q000378:metabolism*; D010957:Plasmids / Q000008:administration & dosage; D017354:Point Mutation; D011233:Precipitin Tests; D017027:Protein Tyrosine Phosphatases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D013997:Time Factors; D015011:Yersinia pseudotuberculosis / Q000737:chemistry*; D000095002:Delta Catenin
D001425:Bacterial Outer Membrane Proteins; D002352:Carrier Proteins; D051177:Catenins; D015815:Cell Adhesion Molecules; D010750:Phosphoproteins; D017027:Protein Tyrosine Phosphatases; C067571:yopH protein, Yersinia; D000095002:Delta Catenin