Influenza virus M2 targets cystic fibrosis transmembrane conductance regulator for lysosomal degradation during viral infection

Londino, J.D.; Lazrak, Ahmed; Noah, James W.; Aggarwal, Saurabh; Bali, Vedrana; Woodworth, Bradford A.; Bebök, Zsuzsanna; Matalon, Sadis

doi:10.1096/fj.14-268755

Cited by 50 publications

(56 citation statements)

References 55 publications

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“…Furthermore, this mechanism of exacerbated oxidative stress and increased production of HA provides possible treatment targets of Cl 2 -induced AHR. Post-Cl 2 -exposure treatment of human airway smooth muscle cells with an antibody against I␣I reverses the L-HA activation of RhoA (22). A previous study has shown increased level of serum urinary trypsin inhibitor, a breakdown product of I␣I in patients with RSV and suggested that this may be a marker of the severity of RSV (40).…”

Section: Discussionmentioning

confidence: 94%

Respiratory syncytial virus infection increases chlorine-induced airway hyperresponsiveness

Song

Doran

et al. 2015

American Journal of Physiology-Lung Cellular and Molecular Phys

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-Exposure to chlorine (Cl2) damages airway and alveolar epithelia resulting in acute lung injury and reactive airway hyperresponsiveness (AHR) to methacholine. However, little is known about the effect of preexisting respiratory disease on Cl2-induced lung injury. By using a murine respiratory syncytial virus (RSV) infection model, we found that preexisting RSV infection increases Cl2 (187 ppm for 30 min)-induced lung inflammation and airway AHR at 24 h after exposure (5 days after infection). RSV infection and Cl2 exposure synergistically induced oxygen desaturation and neutrophil infiltration and increased MCP-1, MIP-1␤, IL-10, IFN-␥, and RANTES concentrations in the bronchoalveolar lavage fluid (BALF). In contrast, levels of type 2 cytokines (i.e., IL-4, IL-5, IL-9, and IL-13) were not significantly affected by either RSV infection or Cl2 exposure. Cl2 exposure, but not RSV infection, induced AHR to methacholine challenge as measured by flexiVent. Moreover, preexisting RSV infection amplified BALF levels of hyaluronan (HA) and AHR. The Cl2-induced AHR was mitigated by treatment with inter-␣-trypsin inhibitor antibody, which inhibits HA signaling, suggesting a mechanism of HA-mediated AHR from exacerbated oxidative injury. Our results show for the first time that preexisting RSV infection predisposes the lung to Cl2-induced injury. These data emphasize the necessity for further research on the effects of Cl2 in vulnerable populations and the development of appropriate treatments. flexiVent; methacholine; bronchoalveolar lavage; cytokines; inflammatory cells CHLORINE (Cl 2 ) is an irritant and reactive gas with significant occupational and environmental hazard concerns due to its wide industrial and domestic usage. Spillage of Cl 2 in the environment during accidents or terrorist acts may result in significant morbidity and mortality to animals and humans (2,15,34,38). Exposure of animals to Cl 2 at concentrations comparable to the vicinity of industrial accidents damages airway epithelia and results in sloughing of airway mucosa, increased alveolar permeability, decreased ability of alveolar epithelial cells to clear fluid, damage to the pulmonary surfactant system (2, 5, 18, 33), coagulation abnormalities (42), and extensive systemic injury (1, 28). Animals surviving the initial exposure show significant structural and functional abnormalities, including the presence of foamy alveolar macrophages, patchy areas of ciliated airway cells (25,26), the presence of excess amounts of goblet cells, hyperplasia and airway hyperreactivity (11), and inability to clear inhaled fungi (15). These injuries may result from chemical reactions of Cl 2 and hypochlorous acid (HOCl), as well as various secondary reaction products, like chloramines (1, 33), chlorinated fatty acid, and low-molecular-weight hyaluronan (HA), a proinflammatory fragment generated by the fractionation of HA (19,21).Currently, little is known about the effects of previous existing respiratory infections on the subsequent Cl 2 -induced lung injury. Herein...

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Section: Discussionmentioning

confidence: 94%

Respiratory syncytial virus infection increases chlorine-induced airway hyperresponsiveness

Song

Doran

et al. 2015

American Journal of Physiology-Lung Cellular and Molecular Phys

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“…To test this hypothesis, we looked at Lys48‐ linked polyubiquitination as a proteasomal degradation signal, in both control and corrector combination‐treated conditions (55). To increase the levels of steady‐state polyubiquitinated CFTR, we inhibited the proteasome with epoxomicin (100 nM; 8 h) (56). The results demonstrated that, in control conditions, the relative polyubiquitination levels of the I507‐ATC ΔF508 CFTR were ~40% lower than those of the I507‐ATT ΔF508 CFTR (Fig.…”

Section: Resultsmentioning

confidence: 99%

A synonymous codon change alters the drug sensitivity of ΔF508 cystic fibrosis transmembrane conductance regulator

et al. 2015

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Synonymous mutations, such as I507-ATC→ATT, in deletion of Phe508 in cystic fibrosis transmembrane conductance regulator (DF508 CFTR), the most frequent disease-associated mutant of CFTR, may affect protein biogenesis, structure, and function and contribute to an altered disease phenotype. Small-molecule drugs are being developed to correct DF508 CFTR. To understand correction mechanisms and the consequences of synonymous mutations, we analyzed the effect of mechanistically distinct correctors, corrector 4a (C4) and lumacaftor (VX-809), on I507-ATT and I507-ATC DF508 CFTR biogenesis and function. C4 stabilized I507-ATT DF508 CFTR band B, but without considerable biochemical and functional correction. VX-809 biochemically corrected ∼10% of both of the variants, leading to stable, forskolin+3-isobutyl-1-methylxanthine (IBMX)-activated whole-cell currents in the presence of the corrector. Omitting VX-809 during whole-cell recordings led to a spontaneous decline of the currents, suggesting posttranslational stabilization by VX-809. Treatment of cells with the C4+VX-809 combination resulted in enhanced rescue and 2-fold higher forskolin+IBMX-activated currents of both I507-ATT and I507-ATC DF508 CFTR, compared with VX-809 treatment alone. The lack of an effect of C4 on I507-ATC DF508 CFTR, but its additive effect in combination with VX-809, implies that C4 acted on VX-809-modified I507-ATC DF508 CFTR. Our results suggest that binding of C4 and VX-809 to DF508 CFTR is conformation specific and provide evidence that synonymous mutations can alter the drug sensitivity of proteins.

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“…As previously described [9], cells were washed three times with PBS pH 8.0, and incubated with EZ-Link Sulfo-NHS-SS-biotin (Thermo Scientific) in PBS at pH 8.0 per manufacturer’s instructions. Cells were then incubated on ice for 15 min, and quenched three times with 50 mM Tris buffer.…”

Section: Methodsmentioning

confidence: 99%

“…The ubiquitination and degradation of membrane proteins is well established and has been demonstrated in receptors such as the interferon alpha receptor, epidermal growth factor receptor (EGFR), Toll-like receptor 2, CFTR, Notch, E-Cadherin, etc. [9–12]. Of the post-translational modifications known to alter protein ubiquitination, phosphorylation is the most widely studied.…”

Section: Introductionmentioning

confidence: 99%

Post-translational modification of the interferon-gamma receptor alters its stability and signaling

Londino

Gulick

Suber

et al. 2017

Biochemical Journal

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The IFN gamma receptor 1 (IFNGR1) binds IFN-γ and activates gene transcription path ways crucial for controlling bacterial and viral infections. Although decreases in IFNGR1 surface levels have been demonstrated to inhibit IFN-γ signaling, little is known regarding the molecular mechanisms controlling receptor stability. Here, we show in epithelial and monocytic cell lines that IFNGR1 displays K48 polyubiquitination, is proteasomally degraded, and harbors three ubiquitin acceptor sites at K277, K279, and K285. Inhibition of glycogen synthase kinase 3 beta (GSK3β) destabilized IFNGR1 while overexpression of GSK3β increased receptor stability. We identified critical serine and threonine residues juxtaposed to ubiquitin acceptor sites that impacted IFNGR1 stability. In CRISPR–Cas9 IFNGR1 generated knockout cell lines, cellular expression of IFNGR1 plasmids encoding ubiquitin acceptor site mutations demonstrated significantly impaired STAT1 phosphoryl ation and decreased STAT1-dependent gene induction. Thus, IFNGR1 undergoes rapid site-specific polyubiquitination, a process modulated by GSK3β. Ubiquitination appears to be necessary for efficient IFNGR1-dependent gamma gene induction and represents a relatively uncharacterized regulatory mechanism for this receptor.

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Influenza virus M2 targets cystic fibrosis transmembrane conductance regulator for lysosomal degradation during viral infection

Cited by 50 publications

References 55 publications

Respiratory syncytial virus infection increases chlorine-induced airway hyperresponsiveness

Respiratory syncytial virus infection increases chlorine-induced airway hyperresponsiveness

A synonymous codon change alters the drug sensitivity of ΔF508 cystic fibrosis transmembrane conductance regulator

Post-translational modification of the interferon-gamma receptor alters its stability and signaling

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