Multiple cyclophilins involved in different cellular pathways mediate HCV replication

Gaither, L. Alex; Borawski, Jason; Anderson, Leah J.; Balabanis, Kara; Dévay, Piroska; Joberty, Gérard; Rau, Christina; Schirle, Markus; Bouwmeester, Tewis; Mickanin, Craig; Zhao, Shan Chuan; Vickers, Chad; Lee, Lac; Deng, Gejing; Baryza, Jeremy; Fujimoto, Roger A.; Lin, Kai; Compton, Teresa; Wiedmann, Brigitte

doi:10.1016/j.virol.2009.10.043

Cited by 78 publications

(101 citation statements)

References 63 publications

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“…Spectra were processed with the Bruker TopSpin software package 1.3 and analyzed using the product plane approach developed in our laboratory (43). Backbone assignments were achieved using two-dimensional 1 H, 15 N HSQC and triple resonance three-dimensional 1 H, 15 N, 13 C HNCO, HN(CA)CO, HNCACB, HN(CO)CACB, and HN(CA)NNH spectra.…”

Section: Methodsmentioning

confidence: 99%

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Domain 3 of NS5A Protein from the Hepatitis C Virus Has Intrinsic α-Helical Propensity and Is a Substrate of Cyclophilin A

Verdegem

Badillo

Wieruszeski

et al. 2011

Journal of Biological Chemistry

101

106

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Nonstructural protein 5A (NS5A) is essential for hepatitis C virus (HCV) replication and constitutes an attractive target for antiviral drug development. Although structural data for its in-plane membrane anchor and domain D1 are available, the structure of domains 2 (D2) and 3 (D3) remain poorly defined. We report here a comparative molecular characterization of the NS5A-D3 domains of the HCV JFH-1 (genotype 2a) and Con1 (genotype 1b) strains. Combining gel filtration, CD, and NMR spectroscopy analyses, we show that NS5A-D3 is natively unfolded. However, NS5A-D3 domains from both JFH-1 and Con1 strains exhibit a propensity to partially fold into an ␣-helix. NMR analysis identifies two putative ␣-helices, for which a molecular model could be obtained. The amphipathic nature of the first helix and its conservation in all genotypes suggest that it might correspond to a molecular recognition element and, as such, promote the interaction with relevant biological partner(s). Because mutations conferring resistance to cyclophilin inhibitors have been mapped into NS5A-D3, we also investigated the functional interaction between NS5A-D3 and cyclophilin A (CypA). CypA indeed interacts with NS5A-D3, and this interaction is completely abolished by cyclosporin A. NMR heteronuclear exchange experiments demonstrate that CypA has in vitro peptidyl-prolyl cis/trans-isomerase activity toward some, but not all, of the peptidyl-prolyl bonds in NS5A-D3. These studies lead to novel insights into the structural features of NS5A-D3 and its relationships with CypA. Hepatitis C virus (HCV),3 a small enveloped virus from the Flaviviridae family, is a major cause of chronic liver disease that may lead to steatosis, liver cirrhosis, and hepatocellular carcinoma. Given about 180 million chronically infected individuals worldwide, HCV is an important health challenge (1). Current therapy is based on a combination of pegylated interferon-␣ and ribavirin but is not fully satisfying because numerous patients are not responding or suffer from serious side effects caused by this treatment. The development of new drugs to treat HCV infections requires a better understanding of the structural and functional features of the viral proteins and their relationships with host cell factors. The HCV genome (ϳ9.6 kb) encodes for a single polyprotein precursor (ϳ3,000 aa) that is co-and post-translationally processed by cellular and viral proteases to yield 10 mature proteins (2, 3). They are classified into structural proteins (Core, E1, and E2), which constitute the viral particle, and nonstructural proteins, of which two, p7 and nonstructural protein 2 (NS2), are required for virus assembly. The remainder of the nonstructural proteins (NS3, NS4A, NS4B, NS5A, and NS5B) are involved in HCV RNA replication (reviewed in Refs. 2 and 3).Cyclophilins are host cell factors that in addition to viral proteins, are equally essential for HCV replication. The human genome encodes up to 16 different cyclophilins (4) that, despite differences in their tissue distributio...

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Section: Methodsmentioning

confidence: 99%

“…However, the precise molecular role of the cyclophilins remains to be elucidated. It has been proposed that CypB, CypA, or even Cyp40 is involved (13)(14)(15)(16)(17). However, recently, several laboratories have identified CypA as the key player in the HCV replication (17)(18)(19)(20).…”

mentioning

confidence: 99%

Domain 3 of NS5A Protein from the Hepatitis C Virus Has Intrinsic α-Helical Propensity and Is a Substrate of Cyclophilin A

Verdegem

Badillo

Wieruszeski

et al. 2011

Journal of Biological Chemistry

101

106

View full text Add to dashboard Cite

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“…We showed that STG-175 has a multi-genotypic activity, exhibits a 13-39 nM EC 50 range among HCV GTs, and does not significantly lose its antiviral activity in the presence of high concentration of human serum. Since we and others showed that host CypA is indispensible for the replication of all HCV GTs [35,[37][38][39][40][41][42], one can speculate that the multi-genotypic activity of STG-175 is due to the fact that it neutralizes the isomerase activity of CypA (Fig 1B).…”

Section: Discussionmentioning

confidence: 82%

“…A difference in the degree of requirement for CypA in the three viral life cycles may also explain the differences in the degree of sensitivity to STG-175 among the three viruses. We and others showed that knocking down CypA by shRNA or siRNA in target cells totally inhibits HCV infection [37][38][40][41] while it partially decreases HIV-1 [66][67][68] and HBV [65] infection. Thus, HCV highly relies on CypA to replicate while HIV-1 and HBV requires CypA to optimally replicate in target cells.…”

Section: Discussionmentioning

confidence: 98%

Characterization of the Anti-HCV Activities of the New Cyclophilin Inhibitor STG-175

Gallay¹,

Chatterji²,

Bobardt³

et al. 2016

Journal of Hepatology

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“…In humans, 19 cyclophilin isoforms with different cellular and tissue distributions have been described (Galat, 2004). Gaither et al (2010) demonstrated that HCV replication relies on several cellular pathways linked to cyclophilins A, H, 40, and E. Cyclosporine A, the cyclophilin inhibitor, was demonstrated to inhibit HCV replication directly, both in vitro and in vivo (Watashi et al, 2003). The finding that cyclosporine A's anti-HCV effect was independent of its immunosuppressive function was promising.…”

Section: Cyclophilin Binding Moleculesmentioning

confidence: 99%

New developments in small molecular compounds for anti-hepatitis C virus (HCV) therapy

Tong

Wang

2012

J. Zhejiang Univ. Sci. B

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Abstract:Infection with hepatitis C virus (HCV) affects approximately 170 million people worldwide. However, no vaccine or immunoglobulin is currently available for the prevention of HCV infection. The standard of care (SOC) involving pegylated interferon-α (PEG-IFN α) plus ribavirin (RBV) for 48 weeks results in a sustained virologic response in less than 50% of patients with chronic hepatitis C genotype 1, the most prevalent type of HCV in North America and Europe. Recently, reliable in vitro culture systems have been developed for accelerating antiviral therapy research, and many new specifically targeted antiviral therapies for hepatitis C (STAT-C) and treatment strategies are being evaluated in clinical trials. These new antiviral agents are expected to improve present treatment significantly and may potentially shorten treatment duration. The aim of this review is to summarize the current developments in new anti-HCV drugs.

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Multiple cyclophilins involved in different cellular pathways mediate HCV replication

Cited by 78 publications

References 63 publications

Domain 3 of NS5A Protein from the Hepatitis C Virus Has Intrinsic α-Helical Propensity and Is a Substrate of Cyclophilin A

Domain 3 of NS5A Protein from the Hepatitis C Virus Has Intrinsic α-Helical Propensity and Is a Substrate of Cyclophilin A

Characterization of the Anti-HCV Activities of the New Cyclophilin Inhibitor STG-175

New developments in small molecular compounds for anti-hepatitis C virus (HCV) therapy

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