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Daclatasvir Prevents Hepatitis C Virus Infectivity by Blocking Transfer of Viral Genome to Assembly Sites
- Source :
- Gastroenterology, Gastroenterology, WB Saunders, 2016, 152 (4), pp.895-907. ⟨10.1053/j.gastro.2016.11.047⟩, Gastroenterology, 2016, 152 (4), pp.895-907. ⟨10.1053/j.gastro.2016.11.047⟩
- Publication Year :
- 2016
- Publisher :
- HAL CCSD, 2016.
-
Abstract
- Daclatasvir is a direct-acting antiviral agent and potent inhibitor of NS5A, which is involved in replication of the hepatitis C virus (HCV) genome, presumably via membranous web shaping, and assembly of new virions, likely via transfer of the HCV RNA genome to viral particle assembly sites. Daclatasvir inhibits the formation of new membranous web structures and, ultimately, of replication complex vesicles, but also inhibits an early assembly step. We investigated the relationship between daclatasvir-induced clustering of HCV proteins, intracellular localization of viral RNAs and inhibition of viral particle assembly.<br />HCVcc particles were produced from Huh7.5 hepatocarcinoma cells in presence of daclatasvir for short time periods. Infectivity as well as production of physical particles were quantified and producer cells were subjected to subcellular fractionation. Intracellular colocalization between core, E2, NS5A, NS4B proteins, and viral RNAs was quantitatively analyzed by confocal microscopy and by structured illumination microscopy.<br />Short exposure of HCV-infected cells to daclatasvir reduced viral assembly and induced clustering of structural proteins with non-structural HCV proteins, including core, E2, NS4B and NS5A. These clustered structures appeared to be inactive assembly platforms, likely owing to loss of functional connection with replication complexes. Daclatasvir greatly reduced delivery of viral genomes to these core clusters without altering HCV RNA colocalization with NS5A. In contrast, daclatasvir neither induced clustered structures nor inhibited HCV assembly in cells infected with a daclatasvir-resistant mutant (NS5A-Y93H), indicating that daclatasvir targets a mutual, specific function of NS5A inhibiting both processes.<br />In addition to inhibiting replication complex biogenesis, daclatasvir prevents viral assembly by blocking transfer of the viral genome to assembly sites. This leads to clustering of HCV proteins, because viral particles and replication complex vesicles cannot form and/or egress. This dual mode of action of daclatasvir could explain its efficacy in blocking HCV replication in cultured cells and in treatment of patients with HCV infection.
- Subjects :
- 0301 basic medicine
Daclatasvir
Pyrrolidines
Hepatitis C virus
viruses
030106 microbiology
Mutant
Genome, Viral
Hepacivirus
Biology
Viral Nonstructural Proteins
medicine.disease_cause
Antiviral Agents
RNA Transport
03 medical and health sciences
Multiplicity of infection
Viral Envelope Proteins
[SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases
Cell Line, Tumor
medicine
Humans
chronic hepatitis C
NS5A
DAA
Infectivity
direct-acting antiviral agent
Hepatology
Endoplasmic reticulum
Viral Core Proteins
Virus Assembly
Gastroenterology
Imidazoles
virus diseases
Valine
[SDV.MHEP.HEG]Life Sciences [q-bio]/Human health and pathology/Hépatology and Gastroenterology
Virology
[SDV.MHEP.HEG] Life Sciences [q-bio]/Human health and pathology/Hépatology and Gastroenterology
digestive system diseases
3. Good health
Protein Transport
030104 developmental biology
[SDV.MHEP.MI] Life Sciences [q-bio]/Human health and pathology/Infectious diseases
RNA, Viral
Carbamates
Biogenesis
medicine.drug
Subjects
Details
- Language :
- English
- ISSN :
- 00165085 and 15280012
- Database :
- OpenAIRE
- Journal :
- Gastroenterology, Gastroenterology, WB Saunders, 2016, 152 (4), pp.895-907. ⟨10.1053/j.gastro.2016.11.047⟩, Gastroenterology, 2016, 152 (4), pp.895-907. ⟨10.1053/j.gastro.2016.11.047⟩
- Accession number :
- edsair.doi.dedup.....fbc5d2d6e13fb403605fb1eb2da1c3f8