title: AMPylation of Rho GTPases subverts multiple host signaling processes.
abstract: Rho GTPases are frequent targets of virulence factors as they are keystone signaling molecules. Herein, we demonstrate that AMPylation of Rho GTPases by VopS is a multifaceted virulence mechanism that counters several host immunity strategies. Activation of NFκB, Erk, and JNK kinase signaling pathways were inhibited in a VopS-dependent manner during infection with Vibrio parahaemolyticus. Phosphorylation and degradation of IKBα were inhibited in the presence of VopS as was nuclear translocation of the NFκB subunit p65. AMPylation also prevented the generation of superoxide by the phagocytic NADPH oxidase complex, potentially by inhibiting the interaction of Rac and p67. Furthermore, the interaction of GTPases with the E3 ubiquitin ligases cIAP1 and XIAP was hindered, leading to decreased degradation of Rac and RhoA during infection. Finally, we screened for novel Rac1 interactions using a nucleic acid programmable protein array and discovered that Rac1 binds to the protein C1QA, a protein known to promote immune signaling in the cytosol. Interestingly, this interaction was disrupted by AMPylation. We conclude that AMPylation of Rho Family GTPases by VopS results in diverse inhibitory consequences during infection beyond the most obvious phenotype, the collapse of the actin cytoskeleton.
authors.affiliation: From the Department of Molecular Biology, UT Southwestern Medical Center, Dallas, Texas 75390-9148 and.
authors.forename: Andrew R
authors.initials: AR
authors.lastname: Woolery
authors.affiliation: The Virginia G. Piper Center for Personalized Diagnostics, Biodesign Institute, Arizona State University, Tempe, Arizona 85287.
authors.forename: Xiaobo
authors.initials: X
authors.lastname: Yu
authors.affiliation: The Virginia G. Piper Center for Personalized Diagnostics, Biodesign Institute, Arizona State University, Tempe, Arizona 85287.
authors.forename: Joshua
authors.initials: J
authors.lastname: LaBaer
authors.affiliation: From the Department of Molecular Biology, UT Southwestern Medical Center, Dallas, Texas 75390-9148 and Kim.Orth@utsouthwestern.edu.
authors.forename: Kim
authors.initials: K
authors.lastname: Orth
mesh_terms: D021581:Active Transport, Cell Nucleus; D000249:Adenosine Monophosphate / Q000378:metabolism; D001426:Bacterial Proteins / Q000378:metabolism*; D002467:Cell Nucleus / Q000378:metabolism; D015922:Complement C1q / Q000378:metabolism; D057809:HEK293 Cells; D054884:Host-Pathogen Interactions; D006801:Humans; D051550:I-kappa B Kinase / Q000378:metabolism; D015151:Immunoblotting; D051034:Inhibitor of Apoptosis Proteins / Q000378:metabolism; D018613:Microscopy, Confocal; D008954:Models, Biological; D010766:Phosphorylation; D011485:Protein Binding; D015398:Signal Transduction; D013481:Superoxides / Q000378:metabolism; D051996:Transcription Factor RelA / Q000378:metabolism; D014736:Vibrio parahaemolyticus / Q000378:metabolism* / Q000502:physiology; D051636:X-Linked Inhibitor of Apoptosis Protein / Q000378:metabolism; D020830:rac1 GTP-Binding Protein / Q000378:metabolism; D020741:rho GTP-Binding Proteins / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D051034:Inhibitor of Apoptosis Proteins; D051996:Transcription Factor RelA; D051636:X-Linked Inhibitor of Apoptosis Protein; D013481:Superoxides; D000249:Adenosine Monophosphate; D015922:Complement C1q; D051550:I-kappa B Kinase; D020830:rac1 GTP-Binding Protein; D020741:rho GTP-Binding Proteins
keywords: AMP; AMPylation; C1qA; GTPase; IAP; NF-κB (NF-KB); Rho (Rho GTPase); RhoGTPases; Signal Transduction; VopS