Intermolecular Binding between TIFA-FHA and TIFA-pT Mediates Tumor Necrosis Factor Alpha Stimulation and NF-κB Activation

Huang, Chia-Chi Flora; Weng, Jui-Hung; Wei, Tong‐You Wade; Wu, Pei; Hsu, Pang-Hung; Chen, Yu-Hou; Wang, Shunchang; Qin, Dongyan; Hung, Chien‐Ching; Chen, Shui-Tsung; Wang, Andrew H.-J.; Shyy, John Y.‐J.; Tsai, Max

doi:10.1128/mcb.00985-12

Cited by 19 publications

(40 citation statements)

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“…Consistent with NF-kB activation, only infection with WT H. pylori, but not TNF-a treatment, induced the formation of so-called TIFAsomes (Gaudet and Gray-Owen, 2016) ( Figure S3A). To further define the importance and specificity of TIFAsome formation upon infection, TIFA-deficient cells were transfected with TIFA-Myc-Flag or a TIFA-Myc-Flag mutant for T9, an important threonine phosphorylation site for TIFA activation (Huang et al, 2012). In cells that remained non-transfected, only TNF-a treatment but not H. pylori infection induced p65 translocation ( Figure 3A, arrowhead in upper panel and lower panel).…”

Section: Role Of Alpk1 In Nf-kb Activationmentioning

confidence: 99%

ALPK1- and TIFA-Dependent Innate Immune Response Triggered by the Helicobacter pylori Type IV Secretion System

et al. 2017

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Activation of transcription factor NF-κB is a hallmark of infection with the gastric pathogen Helicobacter pylori, associated with inflammation and carcinogenesis. Genome-wide RNAi screening revealed numerous host factors involved in H. pylori-, but not IL-1β- and TNF-α-dependent NF-κB regulation. Pathway analysis including CRISPR/Cas9-knockout and recombinant protein technology, immunofluorescence microscopy, immunoblotting, mass spectrometry, and mutant H. pylori strains identified the H. pylori metabolite D-glycero-β-D-manno-heptose 1,7-bisphosphate (βHBP) as a cagPAI type IV secretion system (T4SS)-dependent effector of NF-κB activation in infected cells. Upon pathogen-host cell contact, TIFA forms large complexes (TIFAsomes) including interacting host factors, such as TRAF2. NF-κB activation, TIFA phosphorylation, and TIFAsome formation depend on a functional ALPK1 kinase, highlighting the ALPK1-TIFA axis as a core innate immune pathway. ALPK1-TIFA-mediated NF-κB activation was independent of CagA protein translocation, indicating that CagA translocation and HBP delivery to host cells are distinct features of the pathogen's T4SS.

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Section: Role Of Alpk1 In Nf-kb Activationmentioning

confidence: 99%

ALPK1- and TIFA-Dependent Innate Immune Response Triggered by the Helicobacter pylori Type IV Secretion System

et al. 2017

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“…pcDNA3_TIFA-myc was kindly provided by Jessica Yu (Huang, Weng et al, 2012). The pGEX-14-3-3 constructs GST-14-3-3β, GST-14-3-3ϵ, GST-14-3-3γ, GST-14-3-3η and GST-14-3-3σ were obtained from Cheryl Lyn Walker and Judith Bergeron (University of Texas, USA).…”

Section: Plasmids and Reagentsmentioning

confidence: 99%

XIAP controls RIPK2 signaling by preventing its deposition in speck-like structures

Ellwanger

Arnold

Briese

et al. 2019

Preprint

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The receptor interacting serine/threonine kinase 2 (RIPK2) is essential for linking activation of the pattern recognition receptors NOD1 and NOD2 to cellular signaling events.Recently, it was shown that RIPK2 forms higher order molecular structures in vitro, which were proposed to activate signaling. Here, we demonstrate that RIPK2 forms detergent insoluble complexes in the cytosol of host cells upon infection with invasive enteropathogenic bacteria. Formation of these structures occurred after NF-κB activation and depends on the CARD of NOD1 or NOD2. Complex formation upon activation was dependent on RIPK2 autophosphorylation at Y474 and influenced by phosphorylation at S176. Inhibition of activity of the cIAP protein XIAP induced spontaneous complex formation of RIPK2 but blocked NOD1-dependet NF-κB activation. Using immunoprecipitation, we identified 14-3-3 proteins as novel binding partners of non-activated RIPK2, whereas complexed RIPK2 was bound by the prohibitin proteins Erlin-1 and Erlin-2.Taken together, our work reveals novel roles of XIAP, 14-3-3 and Erlin proteins in the regulation of RIPK2 and expands our knowledge on the function of RIPK2 posttranslational modifications in NOD1/2 signaling. AUTHOR CONTRIBUTIONSCA, KE, SB and IK performed the experiments.JP performed MS analysis and data processing.TAK conceived and supervised the study.CA, KE and TAK wrote and edited the manuscript. CONFLICT OF INTERESTThe authors declare that they have no conflict of interest.A: Indirect immunofluorescence micrographs of HeLa EGFP-RIPK2 cells infected for 2 h with S. flexneri M90T. Staining for XIAP, EGFP-RIPK2, and merge with DNA-staining is shown.Co-localization is shown at higher magnification in the inlay. Scale bar = 10 µm. B: Fluorescence micrographs of HeLa EGFP-RIPK2 cells, treated for 48 h with a XIAP siRNA or a non-targeting siRNA, following infection with S. flexneri M90T for 2 h or left uninfected. Cells were treated with 1 µg/ml doxycycline for 24 h prior infection. EGFP-RIPK2 and merge with DNA-staining is shown. Scale bar = 10 µm. C: IL-8 release in the supernatants from HeLa EGFP-RIPK2 cells, treated for 48 h with a XIAP siRNA or a non-targeting siRNA, following infection with S. flexneri M90T for 6 h. Mean of n=2 with S.D. is shown. D: Immunoblot analysis of HeLa EGFP-RIPK2 cells infected with S. flexneri M90T or BS176 for the indicated time. Immunoblot was probed with anti-RIPK2 antibody, anti-XIAP, and anti-β-tubulin as loading control. E: Left panel: Fluorescence micrographs of HeLa EGFP-RIPK2 cells, transfected with 500 ng Ubi-SMAC-fl-pDS-RED, Ubi-SMAC-tr-pDS-RED, or Ubi-SMAC-AVPI-pDS-RED plasmid and treated with 1 µg/ml doxycycline for 24 h. EGFP-RIPK2, SMAC-dsRed, and merge with DNA-staining is shown. Scale bar = 10 µm. Right panel: Immunoblot analysis of cells prepared as in the left panel. Protein levels were detected using an anti-RIPK2 antibody and anti-β-tubulin as loading control.Figure 6: Phosphorylation of RIPK2 at S176 and Y474 contribute to RIPosome formation. A: Fluorescence microg...

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“…One potential explanation for the disparate functions of TIFA and TIFAB is the requirement of an N-terminal threonine (Thr) residue at position 9 in TIFA. Phosphorylation of Thr-9 on TIFA initiates rapid assembly of a large signaling complex including TIFA and TRAF6, which then elicits downstream signaling (Gaudet et al, 2015;Huang et al, 2012). TIFAB lacks a homologous residue at this position.…”

Section: Introductionmentioning

confidence: 99%

TIFAB Regulates USP15-Mediated p53 Signaling during Stressed and Malignant Hematopoiesis

et al. 2020

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Intermolecular Binding between TIFA-FHA and TIFA-pT Mediates Tumor Necrosis Factor Alpha Stimulation and NF-κB Activation

Cited by 19 publications

References 0 publications

ALPK1- and TIFA-Dependent Innate Immune Response Triggered by the Helicobacter pylori Type IV Secretion System

ALPK1- and TIFA-Dependent Innate Immune Response Triggered by the Helicobacter pylori Type IV Secretion System

XIAP controls RIPK2 signaling by preventing its deposition in speck-like structures

TIFAB Regulates USP15-Mediated p53 Signaling during Stressed and Malignant Hematopoiesis

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