Extended interaction networks with HCV protease NS3-4A substrates explain the lack of adaptive capability against protease inhibitors

Download
2020-10-01
Dultz, Georg
Shimakami, Tetsuro
Schneider, Markus
Murai, Kazuhisa
Yamane, Daisuke
Marıon, Antoıne
Zeitler, Tobias M.
Stross, Claudia
Grimm, Christian
Richter, Rebecca M.
Baeumer, Katrin
Yi, MinKyung
Biondi, Ricardo M.
Zeuzem, Stefan
Tampe, Robert
Antes, Iris
Lange, Christian M.
Welsch, Christoph
Inhibitors against the NS3-4A protease of hepatitis C virus (HCV) have proven to be useful drugs in the treatment of HCV infection. Although variants have been identified with mutations that confer resistance to these inhibitors, the mutations do not restore replicative fitness and no secondary mutations that rescue fitness have been found. To gain insight into the molecular mechanisms underlying the lack of fitness compensation, we screened known resistance mutations in infectious HCV cell culture with different genomic backgrounds. We observed that the Q41R mutation of NS3-4A efficiently rescues the replicative fitness in cell culture for virus variants containing mutations at NS3-Asp(168). To understand how the Q41R mutation rescues activity, we performed protease activity assays complemented by molecular dynamics simulations, which showed that protease-peptide interactions far outside the targeted peptide cleavage sites mediate substrate recognition by NS3-4A and support protease cleavage kinetics. These interactions shed new light on the mechanisms by which NS3-4A cleaves its substrates, viral polyproteins and a prime cellular antiviral adaptor protein, the mitochondrial antiviral signaling protein MAVS. Peptide binding is mediated by an extended hydrogen-bond network in NS3-4A that was effectively optimized for protease-MAVS binding in Asp(168)variants with rescued replicative fitness from NS3-Q41R. In the protease harboring NS3-Q41R, the N-terminal cleavage products of MAVS retained high affinity to the active site, rendering the protease susceptible for potential product inhibition. Our findings reveal delicately balanced protease-peptide interactions in viral replication and immune escape that likely restrict the protease adaptive capability and narrow the virus evolutionary space.
JOURNAL OF BIOLOGICAL CHEMISTRY

Suggestions

Targeting glucosylceramide synthase sensitizes imatinib-resistant chronic myeloid leukemia cells via endogenous ceramide accumulation
BARAN, YUSUF; Bielawski, Jacek; Gündüz, Ufuk; Ogretmen, Besim (2011-10-01)
Purpose Drug resistance presents a major obstacle for the treatment of some patients with chronic myeloid leukemia (CML). Pro-apoptotic ceramide mediates imatinib-induced apoptosis, and metabolism of ceramide by glucosylceramide synthase (GCS) activity, converting ceramide to glucosyl ceramide, might contribute to imatinib resistance. In this study, we investigated the role of ceramide metabolism by GCS in the regulation of imatinib-induced apoptosis in drug-sensitive and drug-resistant K562 and K562/IMA-0....
Upregulation of multi drug resistance genes in doxorubicin resistant human acute myelogeneous leukemia cells and reversal of the resistance
BARAN, YUSUF; Guer, Bala; Kaya, Pelin; Ural, Ali Ugur; Avcu, Ferit; Gündüz, Ufuk (2007-12-01)
The major problem in the treatment of acute myeloid leukemia (AML) patients results from multidrug resistance to administered anticancer agents. Drug resistance proteins, MDR1 and MRP1, which work as drug efflux pumps, can mediate the multidrug resistance of human leukemia cells. In this study, the mechanisms of resistance to doxorubicin-induced cell death in human HL60 AML cells were examined.
Efficient Photodynamic Inactivation of Escherichia coli using a Zinc(II)-Bisdipicolylamine Complex Bearing Porphyrin Derivative and its Liposomal Formulation
Selvi, Hatice Tuğba; Şahin, Sevil; Akbulut, Doğan; Türkyılmaz, Serhan (2021-04-08)
Antibiotic resistant bacterial strains (e.g. MRSA, VRE, MDR-TB and so on) pose a significant threat to public health1 and complete drug resistance could prove to be more lethal than viral pandemics like Covid-19.2 Targeted antibacterial agents may improve antibiotic efficacy compared to non-targeted agents by widening their therapeutic window. While a number of bacterial recognition motifs exist (e.g. antibodies, peptides, glycodendrimers, and cationic dendrimers), zinc(II)-bisdipicolylamine (Zn2BDPA) compl...
Half generations magnetic PAMAM dendrimers as an effective system for targeted gemcitabine delivery
PARSIAN, MARYAM; Mutlu, Pelin; Yalcin, Serap; Tezcaner, Ayşen; Gündüz, Ufuk (2016-12-30)
Tumor-specific delivery of anticancer drugs by magnetic nanoparticles will maximize the efficacy of the drug and minimize side effects, and reduce systemic toxicity. The magnetic core of these nanoparticles provides an advantage for selective drug targeting as they can be targeted to the tumor site and accumulated in cancer cells by means of an external magnetic field. Magnetic nanoparticles can be coated with Polyamidoamine (PAMAM) dendrimer and loaded with drugs. However, biomedical applications of PAMAM ...
Impaired toll like receptor-7 and 9 induced immune activation in chronic spinal cord injured patients contributes to immune dysfunction
Gucluler, Gozde; Adiguzel, Emre; Gungor, Bilgi; Kahraman, Tamer; Gürsel, Mayda; Yilmaz, Bilge; GÜRSEL, İHSAN (Public Library of Science (PLoS), 2017-02-07)
Reduced immune activation or immunosuppression is seen in patients withneurological diseases. Urinary and respiratory infections mainly manifested as septicemia and pneumonia are the most frequent complications following spinal cord injuries and they account for the majority of deaths. The underlying reason of these losses is believed to arise due to impaired immune responses to pathogens. Here, we hypothesized that susceptibility to infections of chronic spinal cord injured (SCI) patients might be due to i...
Citation Formats
G. Dultz et al., “Extended interaction networks with HCV protease NS3-4A substrates explain the lack of adaptive capability against protease inhibitors,” JOURNAL OF BIOLOGICAL CHEMISTRY, pp. 13862–13874, 2020, Accessed: 00, 2021. [Online]. Available: https://hdl.handle.net/11511/90038.