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NS5A inhibitors unmask differences in functional replicase complex half-life between different hepatitis C virus strains
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نویسنده
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benzine t. ,brandt r. ,lovell w.c. ,yamane d. ,neddermann p. ,de francesco r. ,lemon s.m. ,perelson a.s. ,ke r. ,mcgivern d.r.
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منبع
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plos pathogens - 2017 - دوره : 13 - شماره : 6
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چکیده
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Hepatitis c virus (hcv) rna is synthesized by the replicase complex (rc),a macromolecular assembly composed of viral non-structural proteins and cellular co-factors. inhibitors of the hcv ns5a protein block formation of new rcs but do not affect rna synthesis by pre-formed rcs. without new rc formation,existing rcs turn over and are eventually lost from the cell. we aimed to use ns5a inhibitors to estimate the half-life of the functional rc of hcv. we compared different cell culture-infectious strains of hcv that may be grouped based on their sensitivity to lipid peroxidation: robustly replicating,lipid peroxidation resistant (lpor) viruses (e.g. jfh-1 or h77d) and more slowly replicating,lipid peroxidation sensitive (lpos) viruses (e.g. h77s.3 and n.2). in luciferase assays,lposhcv strains declined under ns5a inhibitor therapy with much slower kinetics compared to lporhcv strains. this difference in rate of decline was not observed for inhibitors of the ns5b rna-dependent rna polymerase suggesting that the difference was not simply a consequence of differences in rna stability. in further analyses,we compared two isoclonal hcv variants: the lposh77s.3 and the lporh77d that differ only by 12 amino acids. differences in rate of decline between h77s.3 and h77d following ns5a inhibitor addition were not due to amino acid sequences in ns5a but rather due to a combination of amino acid differences in the non-structural proteins that make up the hcv rc. mathematical modeling of intracellular hcv rna dynamics suggested that differences in rc stability (half-lives of 3.5 and 9.9 hours,for h77d and h77s.3,respectively) are responsible for the different kinetics of antiviral suppression between lposand lporviruses. in nascent rna capture assays,the rate of rna synthesis decline following ns5a inhibitor addition was significantly faster for h77d compared to h77s.3 indicating different half-lives of functional rcs. © 2017 public library of science. all rights reserved.
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آدرس
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lineberger comprehensive cancer center,university of north carolina at chapel hill,chapel hill,nc,united states,division of infectious diseases,department of medicine,university of north carolina at chapel hill,chapel hill,nc, United States, department of mathematics,north carolina state university,raleigh,nc, United States, lineberger comprehensive cancer center,university of north carolina at chapel hill,chapel hill,nc, United States, lineberger comprehensive cancer center,university of north carolina at chapel hill,chapel hill,nc,united states,department of microbiology and cell biology,tokyo metropolitan institute of medical science,setagaya-ku,kamikitazawa,tokyo, Japan, ingm -istituto nazionale di genetica molecolare romeo ed enrica invernizzi,milan,italy,gcp-service international,bremen, Germany, ingm -istituto nazionale di genetica molecolare romeo ed enrica invernizzi,milan, Italy, lineberger comprehensive cancer center,university of north carolina at chapel hill,chapel hill,nc,united states,division of infectious diseases,department of medicine,university of north carolina at chapel hill,chapel hill,nc, United States, theoretical biology and biophysics,los alamos national laboratory,los alamos,nm, United States, department of mathematics,north carolina state university,raleigh,nc, United States, lineberger comprehensive cancer center,university of north carolina at chapel hill,chapel hill,nc,united states,division of infectious diseases,department of medicine,university of north carolina at chapel hill,chapel hill,nc, United States
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Authors
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