HRas Signal Transduction Promotes Hepatitis C Virus Cell Entry by Triggering Assembly of the Host Tetraspanin Receptor Complex

Zona, Laetitia; Lupberger, Joachim; Sidahmed-Adrar, Nazha; Thumann, Christine; Harris, Helen J.; Barnes, Amy; Florentin, Jonathan; Tawar, Rajiv G.; Xiao, Fei; Turek, Marine; Durand, Sarah; Duong, François H. T.; Heim, Markus H.; Cosset, François‐Loïc; Hirsch, Ivan; Samuel, Didier; Brino, Laurent; Zeisel, Mirjam B.; Naour, François Le; McKeating, Jane A.; Baumert, Thomas F.

doi:10.1016/j.chom.2013.02.006

Cited by 148 publications

(205 citation statements)

References 52 publications

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“…Multiple lines of evidence support a critical role for EGFR in HCV entry (38). Binding of HCVcc particles to cells induces EGFR activation (75), EGFR ligands enhance HCV entry (38,75), and EGFR/HRas signaling promotes CD81-CLDN1 binding (41). Moreover, EGFR ligands were shown to increase colocalization of EGFR with CD81 and their localization to early endosomes, suggesting a potential role for EGFR in a step following CD81-CLDN1 engagement (75).…”

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confidence: 99%

“…Additional cellular molecules identified as HCV entry factors include the two receptor tyrosine kinases epidermal growth factor receptor (EGFR) and ephrin type A receptor 2 (EPHA2) (38), the cholesterol uptake molecule Niemann-Pick C1-like 1 (NPC1L1) (39), and transferrin receptor 1 (TFR1) (40). CD81-bound HCV particles have been shown to traffic laterally on the plasma membrane to tight junctions, where they form stable CD81-CLDN1 complexes, and these activities are promoted by protein kinase A (PKA), Rho, and EGFR/HRas signaling (38,(41)(42)(43)(44). Multiple lines of evidence support the finding that HCVpp and HCVcc enter the cell via clathrin-mediated endocytosis (13,(45)(46)(47)(48)(49).…”

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confidence: 99%

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AP-2-Associated Protein Kinase 1 and Cyclin G-Associated Kinase Regulate Hepatitis C Virus Entry and Are Potential Drug Targets

Neveu

Ziv-Av

Barouch-Bentov

et al. 2015

J Virol

121

155

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Hepatitis C virus (HCV) enters its target cell via clathrin-mediated endocytosis. AP-2-associated protein kinase 1 (AAK1) and cyclin G-associated kinase (GAK) are host kinases that regulate clathrin adaptor protein (AP)-mediated trafficking in the endocytic and secretory pathways. We previously reported that AAK1 and GAK regulate HCV assembly by stimulating binding of the subunit of AP-2, AP2M1, to HCV core protein. We also discovered that AAK1 and GAK inhibitors, including the approved anticancer drugs sunitinib and erlotinib, could block HCV assembly. Here, we hypothesized that AAK1 and GAK regulate HCV entry independently of their effect on HCV assembly. Indeed, silencing AAK1 and GAK expression inhibited entry of pseudoparticles and cell culture grown-HCV and internalization of Dil-labeled HCV particles with no effect on HCV attachment or RNA replication. AAK1 or GAK depletion impaired epidermal growth factor (EGF)-mediated enhanced HCV entry and endocytosis of EGF receptor (EGFR), an HCV entry cofactor and erlotinib's cancer target. Moreover, either RNA interference-mediated depletion of AP2M1 or NUMB, each a substrate of AAK1 and/or GAK, or overexpression of either an AP2M1 or NUMB phosphorylation site mutant inhibited HCV entry. Last, in addition to affecting assembly, sunitinib and erlotinib inhibited HCV entry at a postbinding step, their combination was synergistic, and their antiviral effect was reversed by either AAK1 or GAK overexpression. Together, these results validate AAK1 and GAK as critical regulators of HCV entry that function in part by activating EGFR, AP2M1, and NUMB and as the molecular targets underlying the antiviral effect of sunitinib and erlotinib (in addition to EGFR), respectively. IMPORTANCEUnderstanding the host pathways hijacked by HCV is critical for developing host-centered anti-HCV approaches. Entry represents a potential target for antiviral strategies; however, no FDA-approved HCV entry inhibitors are currently available. We reported that two host kinases, AAK1 and GAK, regulate HCV assembly. Here, we provide evidence that AAK1 and GAK regulate HCV entry independently of their role in HCV assembly and define the mechanisms underlying AAK1-and GAK-mediated HCV entry. By regulating temporally distinct steps in the HCV life cycle, AAK1 and GAK represent "master regulators" of HCV infection and potential targets for antiviral strategies. Indeed, approved anticancer drugs that potently inhibit AAK1 or GAK inhibit HCV entry in addition to assembly. These results contribute to an understanding of the mechanisms of HCV entry and reveal attractive host targets for antiviral strategies as well as approved candidate inhibitors of these targets, with potential implications for other viruses that hijack clathrin-mediated pathways. H epatitis C virus (HCV) is a major global health problem, estimated to infect 170 million people worldwide (1, 2). HCV persistence results in severe liver disease, including cirrhosis, liver failure, and hepatocellular carcinoma (reviewed in reference...

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confidence: 99%

mentioning

confidence: 99%

AP-2-Associated Protein Kinase 1 and Cyclin G-Associated Kinase Regulate Hepatitis C Virus Entry and Are Potential Drug Targets

Neveu

Ziv-Av

Barouch-Bentov

et al. 2015

J Virol

121

155

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“…Besides its dipeptidase function, PEPD has been shown to be involved in epidermal growth factor receptor (EGFR) family signaling (37,38). EGFR signaling is believed to play a role during IAV infections (39,40). In addition, gene ontology analysis of the validated hits of the siRNA screens revealed that kinase signaling is the most overrepresented category (31).…”

Section: Resultsmentioning

confidence: 99%

Prolidase Is Required for Early Trafficking Events during Influenza A Virus Entry

Pohl

Edinger

Stertz

2014

J Virol

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Influenza A virus (IAV) entry is a multistep process that requires the interaction of the virus with numerous host factors. In this study, we demonstrate that prolidase (PEPD) is a cellular factor required by IAV for successful entry into target cells. PEPD was selected as a candidate during an entry screen performed on nonvalidated primary hits from previously published genome-wide small interfering RNA (siRNA) screens. siRNA-mediated depletion of PEPD resulted in the decreased growth of IAV during mono-and multicycle growth. This growth defect was independent of cell type or virus strain. Furthermore, IAV restriction was apparent as early as 3 h postinfection, and experiments in the absence of protein biosynthesis revealed that the nuclear import of viral ribonucleoprotein complexes (vRNPs) was already blocked in the absence of PEPD. These results led us to investigate which step during entry was affected. Receptor expression, IAV attachment, or IAV internalization was not dependent on the presence of PEPD. However, when looking at the distribution of incoming IAV particles in PEPD-knockdown cells, we found a localization pattern that differed from that in control cells: IAV mostly localized to the cell periphery, and consequently, viral particles displayed reduced colocalization with early and late endosome markers and fusion between viral and endosomal membranes was strongly reduced. Finally, experiments using a competitive inhibitor of PEPD catalytic activity suggested that the enzymatic function of the dipeptidase is required for its proviral effect on IAV entry. In sum, this study establishes PEPD as a novel entry factor required for early endosomal trafficking of IAV. IMPORTANCEInfluenza A virus (IAV) continues to be a constant threat to public health. As IAV relies on its host cell for replication, the identification of host factors required by the virus is of importance. First, such studies often reveal novel functions of cellular factors and can extend our knowledge of cellular processes. Second, we can further our understanding of processes that are required for the entry of IAV into target cells. Third, the identification of host factors that contribute to IAV entry will increase the number of potential targets for the development of novel antiviral drugs that are of urgent need. Our study identifies prolidase (PEPD) to be a novel entry factor required by IAV for correct routing within the endosomal compartment following virus internalization. Thereby, we link PEPD, which has been shown to play a role during collagen recycling and growth factor signaling, to early events of viral infection.

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“…In vivo treatment of erlotinib resulted in a significant suppression of the viral load in PHH-chimeric mouse model with HCV infection [54]. Furthermore, inhibitors of EGFR downstream kinases Ras (tipifarnib) and Raf (sorafenib) were also assessed and found effective in blocking HCV entry [126].…”

Section: Small Molecules Targeting Host Entry Factors and Cd81-triggementioning

confidence: 99%

Strategies to Preclude Hepatitis C Virus Entry

Burnouf¹,

Lin²

2017

Advances in Treatment of Hepatitis C and B

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Without a preventive vaccine, hepatitis C virus (HCV) remains an important pathogen worldwide with millions of carriers at risk of end-stage liver diseases. Despite the introduction of novel direct-acting antivirals (DAAs), resistance problems, challenges with the difficult-to-treat populations and high costs limit the widespread application of these drugs. Antivirals with alternative mechanism(s) of action, such as by restricting viral entry or cell-to-cell spread, could help expand the scope of antiviral strategies for the management of hepatitis C. Transfusion-associated HCV infection remains another issue in endemic and resource-limited areas around the world. This chapter describes some of the latest developments in antiviral strategies to preclude HCV entry, such as through monoclonal antibodies and small molecules, as well as measures to enhance the safety of therapeutic plasma products in blood transfusion.

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HRas Signal Transduction Promotes Hepatitis C Virus Cell Entry by Triggering Assembly of the Host Tetraspanin Receptor Complex

Cited by 148 publications

References 52 publications

AP-2-Associated Protein Kinase 1 and Cyclin G-Associated Kinase Regulate Hepatitis C Virus Entry and Are Potential Drug Targets

AP-2-Associated Protein Kinase 1 and Cyclin G-Associated Kinase Regulate Hepatitis C Virus Entry and Are Potential Drug Targets

Prolidase Is Required for Early Trafficking Events during Influenza A Virus Entry

Strategies to Preclude Hepatitis C Virus Entry

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