Open AccessDynamics of Subgenomic Hepatitis C Virus Replicon RNA Levels in Huh-7 Cells after Exposure to Nucleoside Antimetabolites
Author(s) -
Lieven Stuyver,
Tamara R. McBrayer,
Phillip M. Tharnish,
Abdalla E. A. Hassan,
Chung K. Chu,
Krzysztof W. Pankiewicz,
K. A. Watanabe,
Raymond F. Schinazi,
Michael Otto
Publication year - 2003
Publication title -
journal of virology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.617
H-Index - 292
eISSN - 1070-6321
pISSN - 0022-538X
DOI - 10.1128/jvi.77.19.10689-10694.2003
Subject(s) - replicon , biology , nucleoside , subgenomic mrna , ribonucleoside , rna , viral replication , cell growth , microbiology and biotechnology , pyrimidine metabolism , cell culture , hepatitis c virus , virology , virus , biochemistry , enzyme , gene , genetics , purine , plasmid
Treatment with antimetabolites results in chemically induced low nucleoside triphosphate pools and cell cycle arrest in exponentially growing cells. Since steady-state levels of hepatitis C virus (HCV) replicon RNA were shown to be dependent on exponential growth of Huh-7 cells, the effects of antimetabolites for several nucleoside biosynthesis pathways on cell growth and HCV RNA levels were investigated. A specific anti-HCV replicon effect was defined as (i). minimal interference with the exponential cell growth, (ii). minimal reduction in cellular host RNA levels, and (iii). reduction of the HCV RNA copy number per cell compared to that of the untreated control. While most antimetabolites caused a cytostatic effect on cell growth, only inhibitors of the de novo pyrimidine ribonucleoside biosynthesis mimicked observations seen in confluent replicon cells, i.e., cytostasis combined with a sharp decrease in replicon copy number per cell. These results suggest that high levels of CTP and UTP are critical parameters for maintaining the steady-state level replication of HCV replicon in Huh-7 cells.