title: Insertion and deletion analyses identify regions of non-structural protein 5A of Hepatitis C virus that are dispensable for viral genome replication.
abstract: Hepatitis C virus (HCV) non-structural protein 5A (NS5A) plays an essential role in viral genome replication. A series of transposon-mediated insertion mutants and deletion mutants of NS5A was used to examine the colony-forming ability of HCV subgenomic replicons encoding the mutant proteins. The results reveal that two regions of NS5A can tolerate insertions: one spanning residues 240-314, which contain the interferon sensitivity-determining region (ISDR), and the other spanning residues 349-417 at the carboxy terminus. The majority of these sites also tolerated insertion of enhanced green fluorescent protein. Furthermore, replicons encoding NS5A with deletions in ISDR or in the carboxy-terminal regions were replication-competent, indicating that these regions of NS5A are not necessary for replication. Taken together, the results suggest that the central region spanning the ISDR and the carboxy-terminal region of the molecule are dispensable for the functions of NS5A in viral genome replication.
authors.affiliation: Department of Veterinary and Biomedical Sciences and Nebraska Center for Virology, University of Nebraska-Lincoln (UNL), E126 Beadle Center, 1901 Vine Street, Lincoln, NE 68588-0666, USA.
authors.forename: Shuanghu
authors.initials: S
authors.lastname: Liu
authors.affiliation: Department of Veterinary and Biomedical Sciences and Nebraska Center for Virology, University of Nebraska-Lincoln (UNL), E126 Beadle Center, 1901 Vine Street, Lincoln, NE 68588-0666, USA.
authors.forename: Israrul H
authors.initials: IH
authors.lastname: Ansari
authors.affiliation: Department of Veterinary and Biomedical Sciences and Nebraska Center for Virology, University of Nebraska-Lincoln (UNL), E126 Beadle Center, 1901 Vine Street, Lincoln, NE 68588-0666, USA.
authors.forename: Subash C
authors.initials: SC
authors.lastname: Das
authors.affiliation: Department of Veterinary and Biomedical Sciences and Nebraska Center for Virology, University of Nebraska-Lincoln (UNL), E126 Beadle Center, 1901 Vine Street, Lincoln, NE 68588-0666, USA.
authors.forename: Asit K
authors.initials: AK
authors.lastname: Pattnaik
mesh_terms: D017353:Gene Deletion; D016679:Genome, Viral; D049452:Green Fluorescent Proteins; D016174:Hepacivirus / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D007372:Interferons / Q000494:pharmacology; D016254:Mutagenesis, Insertional; D017361:Viral Nonstructural Proteins / Q000235:genetics / Q000502:physiology*; D014779:Virus Replication / Q000235:genetics*; D012324:RNA-Dependent RNA Polymerase
chemical_list: D049452:Green Fluorescent Proteins; D007372:Interferons; D017361:Viral Nonstructural Proteins; C087036:NS-5 protein, hepatitis C virus; D012324:RNA-Dependent RNA Polymerase