1. Biochemistry and Chemical Biology
  2. Cell Biology

INAVA-ARNO complexes bridge mucosal barrier function with inflammatory signaling

  1. Phi H Luong
  2. Matija Hedl
  3. Jie Yan
  4. Tao Zuo
  5. Tian-Min Fu
  6. Xiaomo Jiang
  7. Jay R Thiagarajah
  8. Steen H Hansen
  9. Cammie F Lesser
  10. Hao Wu
  11. Clara Abraham
  12. Wayne I Lencer  Is a corresponding author
  1. Harvard Medical School, United States
  2. Yale University, United States
  3. Novartis Institutes for Biomedical Research, United States
  4. Massachusetts General Hospital, United States
https://doi.org/10.7554/eLife.38539
Share this article
Cite this article
  1. Phi H Luong
  2. Matija Hedl
  3. Jie Yan
  4. Tao Zuo
  5. Tian-Min Fu
  6. Xiaomo Jiang
  7. Jay R Thiagarajah
  8. Steen H Hansen
  9. Cammie F Lesser
  10. Hao Wu
  11. Clara Abraham
  12. Wayne I Lencer
(2018)
INAVA-ARNO complexes bridge mucosal barrier function with inflammatory signaling
eLife 7:e38539.
https://doi.org/10.7554/eLife.38539
  2. Download BibTeX
  3. Download .RIS

Abstract

Homeostasis at mucosal surfaces requires cross-talk between the environment and barrier epithelial cells. Disruption of barrier function typifies mucosal disease. Here we elucidate a bifunctional role in coordinating this cross-talk for the inflammatory bowel disease risk-gene INAVA. Both activities require INAVA's DUF3338 domain (renamed CUPID). CUPID stably binds the cytohesin ARF-GEF ARNO to effect lateral membrane F-actin assembly underlying cell-cell junctions and barrier function. Unexpectedly, when bound to CUPID, ARNO affects F-actin dynamics in the absence of its canonical activity as a guanine nucleotide-exchange factor. Upon exposure to IL-1β, INAVA relocates to form cytosolic puncta, where CUPID amplifies TRAF6-dependent polyubiquitination and inflammatory signaling. In this case, ARNO binding to CUPID negatively-regulates polyubiquitination and the inflammatory response. INAVA and ARNO act similarly in primary human macrophages responding to IL-1β and NOD2 agonists. Thus, INAVA-CUPID exhibits dual functions, coordinated directly by ARNO, that bridge epithelial barrier function with extracellular signals and inflammation.

Data availability

All data analysed during this study are included in the manuscript. Source data have been provided for Figure 3 C-F.

Article and author information

Author details

  1. Phi H Luong

    Department of Pediatrics, Harvard Medical School, Boston, United States
    Competing interests
    No competing interests declared.

  2. Matija Hedl

    Department of Medicine, Yale University, New Haven, United States
    Competing interests
    No competing interests declared.

  3. Jie Yan

    Department of Medicine, Yale University, New Haven, United States
    Competing interests
    No competing interests declared.

  4. Tao Zuo

    Department of Pediatrics, Harvard Medical School, Boston, United States
    Competing interests
    No competing interests declared.

  5. Tian-Min Fu

    Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, United States
    Competing interests
    No competing interests declared.

  6. Xiaomo Jiang

    Novartis Institutes for Biomedical Research, Cambridge, United States
    Competing interests
    Xiaomo Jiang, is affiliated with Novartis. The author has no other competing interests to declare.

  7. Jay R Thiagarajah

    Department of Pediatrics, Harvard Medical School, Boston, United States
    Competing interests
    No competing interests declared.

  8. Steen H Hansen

    Department of Pediatrics, Harvard Medical School, Boston, United States
    Competing interests
    No competing interests declared.

  9. Cammie F Lesser

    Department of Medicine, Division of Infectious Diseases, Massachusetts General Hospital, Cambridge, United States
    Competing interests
    No competing interests declared.

  10. Hao Wu

    Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, United States
    Competing interests
    No competing interests declared.

  11. Clara Abraham

    Department of Medicine, Yale University, New Haven, United States
    Competing interests
    No competing interests declared.

  12. Wayne I Lencer

    Department of Pediatrics, Harvard Medical School, Boston, United States
    For correspondence
    Wayne.Lencer@childrens.harvard.edu
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-7346-2730

Funding

National Institutes of Health (T32HD007466)

  • Phi H Luong

Crohn's and Colitis Foundation of America (Career Development Award)

  • Phi H Luong

Boston Children's Hospital (Rubin-Wolpow Fellowship)

  • Phi H Luong

National Institutes of Health (DK099097)

  • Clara Abraham

National Institutes of Health (DK048106)

  • Wayne I Lencer

National Institutes of Health (DK084424)

  • Wayne I Lencer

National Institutes of Health (P30 DK034854)

  • Wayne I Lencer

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Copyright

© 2018, Luong et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

Metrics

  • 1,552
    views
  • 278
    downloads
  • 18
    citations

Views, downloads and citations are aggregated across all versions of this paper published by eLife.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Phi H Luong
  2. Matija Hedl
  3. Jie Yan
  4. Tao Zuo
  5. Tian-Min Fu
  6. Xiaomo Jiang
  7. Jay R Thiagarajah
  8. Steen H Hansen
  9. Cammie F Lesser
  10. Hao Wu
  11. Clara Abraham
  12. Wayne I Lencer
(2018)
INAVA-ARNO complexes bridge mucosal barrier function with inflammatory signaling
eLife 7:e38539.
https://doi.org/10.7554/eLife.38539

Share this article

https://doi.org/10.7554/eLife.38539

Categories and tags

Research organism

Further reading

    1. Biochemistry and Chemical Biology
    2. Microbiology and Infectious Disease

    On the role of VP3-PI3P interaction in birnavirus endosomal membrane targeting

    Flavia A Zanetti, Ignacio Fernandez ... Laura Ruth Delgui
    Research Article

    Birnaviruses are a group of double-stranded RNA (dsRNA) viruses infecting birds, fish, and insects. Early endosomes (EE) constitute the platform for viral replication. Here, we study the mechanism of birnaviral targeting of EE membranes. Using the Infectious Bursal Disease Virus (IBDV) as a model, we validate that the viral protein 3 (VP3) binds to phosphatidylinositol-3-phosphate (PI3P) present in EE membranes. We identify the domain of VP3 involved in PI3P-binding, named P2 and localized in the core of VP3, and establish the critical role of the arginine at position 200 (R200), conserved among all known birnaviruses. Mutating R200 abolishes viral replication. Moreover, we propose a two-stage modular mechanism for VP3 association with EE. Firstly, the carboxy-terminal region of VP3 adsorbs on the membrane, and then the VP3 core reinforces the membrane engagement by specifically binding PI3P through its P2 domain, additionally promoting PI3P accumulation.

    1. Biochemistry and Chemical Biology
    2. Microbiology and Infectious Disease

    Cellular Energy Production: Mycofactocin and the mycobacterial electron transport chain

    Stephanie M Stuteley, Ghader Bashiri
    Insight

    In the bacterium M. smegmatis, an enzyme called MftG allows the cofactor mycofactocin to transfer electrons released during ethanol metabolism to the electron transport chain.

    1. Biochemistry and Chemical Biology
    2. Structural Biology and Molecular Biophysics

    A conformational fingerprint for amyloidogenic light chains

    Cristina Paissoni, Sarita Puri ... Carlo Camilloni
    Research Article

    Both immunoglobulin light-chain (LC) amyloidosis (AL) and multiple myeloma (MM) share the overproduction of a clonal LC. However, while LCs in MM remain soluble in circulation, AL LCs misfold into toxic-soluble species and amyloid fibrils that accumulate in organs, leading to distinct clinical manifestations. The significant sequence variability of LCs has hindered the understanding of the mechanisms driving LC aggregation. Nevertheless, emerging biochemical properties, including dimer stability, conformational dynamics, and proteolysis susceptibility, distinguish AL LCs from those in MM under native conditions. This study aimed to identify a2 conformational fingerprint distinguishing AL from MM LCs. Using small-angle X-ray scattering (SAXS) under native conditions, we analyzed four AL and two MM LCs. We observed that AL LCs exhibited a slightly larger radius of gyration and greater deviations from X-ray crystallography-determined or predicted structures, reflecting enhanced conformational dynamics. SAXS data, integrated with molecular dynamics simulations, revealed a conformational ensemble where LCs adopt multiple states, with variable and constant domains either bent or straight. AL LCs displayed a distinct, low-populated, straight conformation (termed H state), which maximized solvent accessibility at the interface between constant and variable domains. Hydrogen-deuterium exchange mass spectrometry experimentally validated this H state. These findings reconcile diverse experimental observations and provide a precise structural target for future drug design efforts.