PMID- 25666154
OWN - NLM
STAT- MEDLINE
DCOM- 20160111
LR  - 20220110
IS  - 1098-6596 (Electronic)
IS  - 0066-4804 (Print)
IS  - 0066-4804 (Linking)
VI  - 59
IP  - 5
DP  - 2015 May
TI  - Cyclophilin and NS5A inhibitors, but not other anti-hepatitis C virus (HCV) 
      agents, preclude HCV-mediated formation of double-membrane-vesicle viral 
      factories.
PG  - 2496-507
LID - 10.1128/AAC.04958-14 [doi]
AB  - Although the mechanisms of action (MoA) of nonstructural protein 3 inhibitors 
      (NS3i) and NS5B inhibitors (NS5Bi) are well understood, the MoA of cyclophilin 
      inhibitors (CypI) and NS5A inhibitors (NS5Ai) are not fully defined. In this 
      study, we examined whether CypI and NS5Ai interfere with hepatitis C virus (HCV) 
      RNA synthesis of replication complexes (RCs) or with an earlier step of HCV RNA 
      replication, the creation of double-membrane vesicles (DMVs) essential for HCV 
      RNA replication. In contrast to NS5Bi, both CypI and NS5Ai do not block HCV RNA 
      synthesis by way of RCs, suggesting that they exert their antiviral activity 
      prior to the establishment of enzymatically active RCs. We found that viral 
      replication is not a precondition for DMV formation, since the NS3-NS5B 
      polyprotein or NS5A suffices to create DMVs. Importantly, only CypI and NS5Ai, 
      but not NS5Bi, mir-122, or phosphatidylinositol-4 kinase IIIalpha (PI4KIIIalpha) 
      inhibitors, prevent NS3-NS5B-mediated DMV formation. NS3-NS5B was unable to 
      create DMVs in cyclophilin A (CypA) knockdown (KD) cells. We also found that the 
      isomerase activity of CypA is absolutely required for DMV formation. This not 
      only suggests that NS5A and CypA act in concert to build membranous viral 
      factories but that CypI and NS5Ai mediate their early anti-HCV effects by 
      preventing the formation of organelles, where HCV replication is normally 
      initiated. This is the first investigation to examine the effect of a large panel 
      of anti-HCV agents on DMV formation, and the results reveal that CypI and NS5Ai 
      act at the same membranous web biogenesis step of HCV RNA replication, thus 
      indicating a new therapeutic target of chronic hepatitis C.
CI  - Copyright (c) 2015, American Society for Microbiology. All Rights Reserved.
FAU - Chatterji, Udayan
AU  - Chatterji U
AD  - Department of Immunology & Microbial Science, The Scripps Research Institute, La 
      Jolla, California, USA.
FAU - Bobardt, Michael
AU  - Bobardt M
AD  - Department of Immunology & Microbial Science, The Scripps Research Institute, La 
      Jolla, California, USA.
FAU - Tai, Andrew
AU  - Tai A
AD  - Division of Gastroenterology, Department of Internal Medicine, University of 
      Michigan, Ann Arbor, Michigan, USA.
FAU - Wood, Malcolm
AU  - Wood M
AD  - Department of Immunology & Microbial Science, The Scripps Research Institute, La 
      Jolla, California, USA.
FAU - Gallay, Philippe A
AU  - Gallay PA
AD  - Department of Immunology & Microbial Science, The Scripps Research Institute, La 
      Jolla, California, USA gallay@scripps.edu.
LA  - eng
GR  - R01 AI087746/AI/NIAID NIH HHS/United States
GR  - R01 AI105007/AI/NIAID NIH HHS/United States
GR  - R01 DK097374/DK/NIDDK NIH HHS/United States
GR  - AI087746/AI/NIAID NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
DEP - 20150209
PL  - United States
TA  - Antimicrob Agents Chemother
JT  - Antimicrobial agents and chemotherapy
JID - 0315061
RN  - 0 (Antiviral Agents)
RN  - 0 (Viral Nonstructural Proteins)
RN  - EC 2.7.7.48 (NS-5 protein, hepatitis C virus)
RN  - EC 5.2.1.- (Cyclophilins)
SB  - IM
MH  - Antiviral Agents/*pharmacology
MH  - Cell Line, Tumor
MH  - Cyclophilins/*antagonists & inhibitors
MH  - Hepacivirus/*drug effects/metabolism
MH  - Humans
MH  - Viral Nonstructural Proteins/*antagonists & inhibitors
MH  - Virus Replication/drug effects
PMC - PMC4394764
EDAT- 2015/02/11 06:00
MHDA- 2016/01/12 06:00
PMCR- 2015/11/01
CRDT- 2015/02/11 06:00
PHST- 2014/12/07 00:00 [received]
PHST- 2015/02/02 00:00 [accepted]
PHST- 2015/02/11 06:00 [entrez]
PHST- 2015/02/11 06:00 [pubmed]
PHST- 2016/01/12 06:00 [medline]
PHST- 2015/11/01 00:00 [pmc-release]
AID - AAC.04958-14 [pii]
AID - 04958-14 [pii]
AID - 10.1128/AAC.04958-14 [doi]
PST - ppublish
SO  - Antimicrob Agents Chemother. 2015 May;59(5):2496-507. doi: 10.1128/AAC.04958-14. 
      Epub 2015 Feb 9.