SARS-CoV proteins decrease levels and activity of human ENaC via activation of distinct PKC isoforms
Department of Anesthesiology, University of Alabama at Birmingham School of Medicine, Birmingham, Alabama 35233-6810, USA. AJP Lung Cellular and Molecular Physiology
(Impact Factor: 4.08).
01/2009; 296(3):L372-83. DOI: 10.1152/ajplung.90437.2008
Among the multiple organ disorders caused by the severe acute respiratory syndrome coronavirus (SARS-CoV), acute lung failure following atypical pneumonia is the most serious and often fatal event. We hypothesized that two of the hydrophilic structural coronoviral proteins (S and E) would regulate alveolar fluid clearance by decreasing the cell surface expression and activity of amiloride-sensitive epithelial sodium (Na(+)) channels (ENaC), the rate-limiting protein in transepithelial Na(+) vectorial transport across distal lung epithelial cells. Coexpression of either S or E protein with human alpha-, beta-, and gamma-ENaC in Xenopus oocytes led to significant decreases of both amiloride-sensitive Na(+) currents and gamma-ENaC protein levels at their plasma membranes. S and E proteins decreased the rate of ENaC exocytosis and either had no effect (S) or decreased (E) rates of endocytosis. No direct interactions among SARS-CoV E protein with either alpha- or gamma-ENaC were indentified. Instead, the downregulation of ENaC activity by SARS proteins was partially or completely restored by administration of inhibitors of PKCalpha/beta1 and PKCzeta. Consistent with the whole cell data, expression of S and E proteins decreased ENaC single-channel activity in oocytes, and these effects were partially abrogated by PKCalpha/beta1 inhibitors. Finally, transfection of human airway epithelial (H441) cells with SARS E protein decreased whole cell amiloride-sensitive currents. These findings indicate that lung edema in SARS infection may be due at least in part to activation of PKC by SARS proteins, leading to decreasing levels and activity of ENaC at the apical surfaces of lung epithelial cells.
Available from: Michael Kashon
- "The PCL is maintained by the coordinated action of many ion channels, pumps, and transporters (Knowles et al. 1984; Toczylowska-Maminska and Dolowy 2012). Disruption of ion transport can contribute to airway diseases, such as mucus thickening in cystic fibrosis due to PCL dehydration , lung edema due to an inhibition of epithelial Na + channels (ENaC; Chen et al. 2004; Morty et al. 2007; Ji et al. 2009), and interfere with regulatory mechanisms in the airways, such as the release of epithelium-derived relaxing factor which induces relaxation of airway smooth muscle and submucosal blood vessels (Prazma et al. 1994; Fedan et al. 2004; Wu et al. 2004). There is a large body of evidence supporting the notion that NGF can alter ion transport in nonpulmonary cells. "
[Show abstract] [Hide abstract]
ABSTRACT: Nerve growth factor (NGF) is overexpressed in patients with inflammatory lung diseases, including virus infections. Airway surface liquid (ASL), which is regulated by epithelial cell ion transport, is essential for normal lung function. No information is available regarding the effect of NGF on ion transport of airway epithelium. To investigate whether NGF can affect ion transport, human primary air-interface cultured epithelial cells were placed in Ussing chambers to obtain transepithelial voltage (−7.1 ± 3.4 mV), short-circuit current (Isc, 5.9 ± 1.0 μA), and transepithelial resistance (750 Ω·cm2), and to measure responses to ion transport inhibitors. Amiloride (apical, 3.5 × 10−5 mol/L) decreased Isc by 55.3%. Apically applied NGF (1 ng/mL) reduced Isc by 5.3% in 5 min; basolaterally applied NGF had no effect. The response to amiloride was reduced (41.6%) in the presence of NGF. K-252a (10 nmol/L, apical) did not itself affect Na+ transport, but it attenuated the NGF-induced reduction in Na+ transport, indicating the participation of the trkA receptor in the NGF-induced reduction in Na+ transport. PD-98059 (30 μmol/L, apical and basolateral) did not itself affect Na+ transport, but attenuated the NGF-induced reduction in Na+ transport, indicating that trkA activated the Erk 1/2 signaling cascade. NGF stimulated phosphorylation of Erk 1/2 and the β-subunit of ENaC. K-252a and PD-98059 inhibited these responses. NGF had no effect on Isc in the presence of apical nystatin (50 μmol/L). These results indicate that NGF inhibits Na+ transport through a trkA-Erk 1/2-activated signaling pathway linked to ENaC phosphorylation.
07/2014; 2(7). DOI:10.14814/phy2.12073
Available from: Jose Angel Regla-Nava
- "Possibly, these interactions could lead to partial relocalization of these proteins and therefore to the observed reduction of ion transport, which may explain decreased voltage activated currents intensity detected by whole-cell patch clamp assays. Importantly, in agreement with our work, it has been shown that E protein does not display ion channel activity in a different experimental system, the plasma membrane of Xenopus oocytes (Ji et al., 2009) but decreases the levels and activity of human epithelial sodium channels, affecting ion transport at the plasma membrane level. Overall, our results from four complementary approaches strongly suggest that SARS-CoV E protein did not accumulate at the plasma membrane during SARS-CoV infection, or when expressed alone, suggesting that the intrinsic activities of E protein should be displayed in the inner organelles and that plasma membrane permeabilization to monovalent ions (Pervushin et al., 2009) or to small weight compounds in E protein expressing cells (Liao et al., 2004, 2006; Madan et al., 2008), is most likely due to indirect effects. "
[Show abstract] [Hide abstract]
ABSTRACT: Severe acute respiratory syndrome (SARS) coronavirus (CoV) envelope (E) protein is a transmembrane protein. Several subcellular locations and topological conformations of E protein have been proposed. To identify the correct ones, polyclonal and monoclonal antibodies specific for the amino or the carboxy terminus of E protein, respectively, were generated. E protein was mainly found in the endoplasmic reticulum-Golgi intermediate compartment (ERGIC) of cells transfected with a plasmid encoding E protein or infected with SARS-CoV. No evidence of E protein presence in the plasma membrane was found by using immunofluorescence, immunoelectron microscopy and cell surface protein labeling. In addition, measurement of plasma membrane voltage gated ion channel activity by whole-cell patch clamp suggested that E protein was not present in the plasma membrane. A topological conformation in which SARS-CoV E protein amino terminus is oriented towards the lumen of intracellular membranes and carboxy terminus faces cell cytoplasm is proposed.
Virology 07/2011; 415(2):69-82. DOI:10.1016/j.virol.2011.03.029 · 3.32 Impact Factor
Available from: Jürg Hamacher
- "PKC is an important negative regulator of ENaC expression. Recently PKC alpha (PKC-α) and zeta isozymes were found to be crucial in ENaC downregulation caused by proteins of SARS-CoV17 and of Influenza A virus18. Recently, also oxidative stress, which often occurs in the lung under conditions such as infection and inflammation, has been demonstrated to interfere with ENaC expression. "
[Show abstract] [Hide abstract]
ABSTRACT: —Pulmonary edema, a major manifestation of left ventricular heart failure, renal insufficiency, shock, diffuse alveolar damage and lung hypersensitivity states, is a significant medical problem worldwide and can be life-threatening. The proinflammatory cytokine tumor necrosis factor (TNF) has been shown to contribute to the pathogenesis and development of pulmonary edema. However, some recent studies have demonstrated surprisingly that TNF can also promote alveolar fluid reabsorption in vivo and in vitro. This protective effect of the cytokine is mediated by the lectin-like domain of the cytokine, which is spatially distinct from the TNF receptor binding sites. The TIP peptide, a synthetic mimic of the lectin-like domain of TNF, can significantly increase alveolar fluid clearance and improve lung compliance in pulmonary edema models. In this review, we will discuss the dual role of TNF in pulmonary edema.
—tumor necrosis factor (TNF); acute lung injury (ALI); acute respiratory distress syndrome (ARDS); positive end-expiratory pressure (PEEP);epithelial sodium channel (ENaC);neural precursor cell-expressed developmentally downregulated (gene 4) protein (Nedd4-2);serum and glucocorticoid dependent kinase (Sgk-1);insulin-like growth factor 1 (IGF-1);Protein Kinase C (PKC);reactive oxygen species (ROS);myosin light chain (MLC);pneumolysin (PLY);listeriolysin (LLO);interleukin (IL);bronchoalveolar lavage fluids (BALF);Bacillus Calmette-Guerin (BCG);TNF receptor type 1 (TNFR1); TNF receptor type 2 (TNF-R2);
Journal of cardiovascular disease research 03/2010; 1(1):29-36. DOI:10.4103/0975-3583.59983
Data provided are for informational purposes only. Although carefully collected, accuracy cannot be guaranteed. The impact factor represents a rough estimation of the journal's impact factor and does not reflect the actual current impact factor. Publisher conditions are provided by RoMEO. Differing provisions from the publisher's actual policy or licence agreement may be applicable.