Cytotoxic necrotizing factor-Y boosts Yersinia effector translocation by activating Rac protein

TY - JOUR

T1 - Cytotoxic necrotizing factor-Y boosts Yersinia effector translocation by activating Rac protein

AU - Wolters, Manuel

AU - Boyle, Colleen Erin

AU - Lardong, Kerstin

AU - Trülzsch, Konrad

AU - Steffen, Anika

AU - Rottner, Klemens

AU - Ruckdeschel, Klaus

AU - Aepfelbacher, Martin

PY - 2013/8/9

Y1 - 2013/8/9

N2 - Pathogenic Yersinia spp. translocate the effectors YopT, YopE, and YopO/YpkA into target cells to inactivate Rho family GTP-binding proteins and block immune responses. Some Yersinia spp. also secrete the Rho protein activator cytotoxic necrotizing factor-Y (CNF-Y), but it has been unclear how the bacteria may benefit from Rho protein activation. We show here that CNF-Y increases Yop translocation in Yersinia enterocolitica-infected cells up to 5-fold. CNF-Y strongly activated RhoA and also delayed in time Rac1 and Cdc42, but when individually expressed, constitutively active mutants of Rac1, but not of RhoA, increased Yop translocation. Consistently, knock-out or knockdown of Rac1 but not of RhoA, -B, or -C inhibited Yersinia effector translocation in CNF-Y-treated and control cells. Activation or knockdown of Cdc42 also affected Yop translocation but much less efficiently than Rac. The increase in Yop translocation induced by CNF-Y was essentially independent of the presence of YopE, YopT, or YopO in the infecting Yersinia strain, indicating that none of the Yops reported to inhibit translocation could reverse the CNF-Y effect. In summary, the CNF-Y activity of Yersinia strongly enhances Yop translocation through activation of Rac.

AB - Pathogenic Yersinia spp. translocate the effectors YopT, YopE, and YopO/YpkA into target cells to inactivate Rho family GTP-binding proteins and block immune responses. Some Yersinia spp. also secrete the Rho protein activator cytotoxic necrotizing factor-Y (CNF-Y), but it has been unclear how the bacteria may benefit from Rho protein activation. We show here that CNF-Y increases Yop translocation in Yersinia enterocolitica-infected cells up to 5-fold. CNF-Y strongly activated RhoA and also delayed in time Rac1 and Cdc42, but when individually expressed, constitutively active mutants of Rac1, but not of RhoA, increased Yop translocation. Consistently, knock-out or knockdown of Rac1 but not of RhoA, -B, or -C inhibited Yersinia effector translocation in CNF-Y-treated and control cells. Activation or knockdown of Cdc42 also affected Yop translocation but much less efficiently than Rac. The increase in Yop translocation induced by CNF-Y was essentially independent of the presence of YopE, YopT, or YopO in the infecting Yersinia strain, indicating that none of the Yops reported to inhibit translocation could reverse the CNF-Y effect. In summary, the CNF-Y activity of Yersinia strongly enhances Yop translocation through activation of Rac.

KW - Animals

KW - Bacterial Outer Membrane Proteins

KW - Bacterial Toxins

KW - HeLa Cells

KW - Humans

KW - Mice

KW - Mice, Knockout

KW - Neuropeptides

KW - Protein Transport

KW - Yersinia Infections

KW - Yersinia enterocolitica

KW - cdc42 GTP-Binding Protein

KW - rac1 GTP-Binding Protein

KW - ras Proteins

KW - rho GTP-Binding Proteins

KW - rhoA GTP-Binding Protein

KW - rhoB GTP-Binding Protein

U2 - 10.1074/jbc.M112.448662

DO - 10.1074/jbc.M112.448662

M3 - SCORING: Journal article

C2 - 23803609

VL - 288

SP - 23543

EP - 23553

JO - J BIOL CHEM

JF - J BIOL CHEM

SN - 0021-9258

IS - 32

ER -