4OBC

Crystal structure of HCV polymerase NS5b genotype 2a JFH-1 isolate with the S15G, C223H, V321I resistance mutations against the guanosine analog GS-0938 (PSI-3529238)

Summary for 4OBC

• Entry DOI: 10.2210/pdb4obc/pdb
• Related: 4E76 4E78 4E7A
• Descriptor: RNA-directed RNA polymerase, 2-(N-MORPHOLINO)-ETHANESULFONIC ACID, GLYCEROL, ... (6 entities in total)
• Functional Keywords: hepatitis, hcv, viral polymerase, rna-dependent-rna-polymerase, rdrp, resistance, nucleotide analog inhibitor, viral protein, transferase
• Biological source: Hepatitis C virus JFH-1 (HCV)
• Cellular location: Core protein p21: Host endoplasmic reticulum membrane ; Single-pass membrane protein . Core protein p19: Virion . Envelope glycoprotein E1: Virion membrane ; Single-pass type I membrane protein . Envelope glycoprotein E2: Virion membrane ; Single-pass type I membrane protein . p7: Host endoplasmic reticulum membrane ; Multi-pass membrane protein . Protease NS2-3: Host endoplasmic reticulum membrane ; Multi-pass membrane protein . Serine protease NS3: Host endoplasmic reticulum membrane ; Peripheral membrane protein . Non-structural protein 4A: Host endoplasmic reticulum membrane ; Single-pass type I membrane protein . Non-structural protein 4B: Host endoplasmic reticulum membrane ; Multi-pass membrane protein . Non-structural protein 5A: Host endoplasmic reticulum membrane ; Peripheral membrane protein . RNA-directed RNA polymerase: Host endoplasmic reticulum membrane ; Single-pass type I membrane protein : Q99IB8
• Total number of polymer chains: 1
• Total formula weight: 65500.59

Authors

Edwards, T.E.,Abendroth, J.,Appleby, T.C. (deposition date: 2014-01-07, release date: 2014-09-10, Last modification date: 2023-09-20)

Primary citation

Lam, A.M.,Edwards, T.E.,Mosley, R.T.,Murakami, E.,Bansal, S.,Lugo, C.,Bao, H.,Otto, M.J.,Sofia, M.J.,Furman, P.A.
Molecular and Structural Basis for the Roles of Hepatitis C Virus Polymerase NS5B Amino Acids 15, 223, and 321 in Viral Replication and Drug Resistance.
Antimicrob.Agents Chemother., 58:6861-6869, 2014

PubMed Abstract: Resistance to the 2'-F-2'-C-methylguanosine monophosphate nucleotide hepatitis C virus (HCV) inhibitors PSI-352938 and PSI-353661 was associated with a combination of amino acid changes (changes of S to G at position 15 [S15G], C223H, and V321I) within the genotype 2a nonstructural protein 5B (NS5B), an RNA-dependent RNA polymerase. To understand the role of these residues in viral replication, we examined the effects of single and multiple point mutations on replication fitness and inhibition by a series of nucleotide analog inhibitors. An acidic residue at position 15 reduced replicon fitness, consistent with its proximity to the RNA template. A change of the residue at position 223 to an acidic or large residue reduced replicon fitness, consistent with its proposed proximity to the incoming nucleoside triphosphate (NTP). A change of the residue at position 321 to a charged residue was not tolerated, consistent with its position within a hydrophobic cavity. This triple resistance mutation was specific to both genotype 2a virus and 2'-F-2'-C-methylguanosine inhibitors. A crystal structure of the NS5B S15G/C223H/V321I mutant of the JFH-1 isolate exhibited rearrangement to a conformation potentially consistent with short primer-template RNA binding, which could suggest a mechanism of resistance accomplished through a change in the NS5B conformation, which was better tolerated by genotype 2a virus than by viruses of other genotypes.

PubMed: 25182647
DOI: 10.1128/AAC.03847-14

Experimental method

X-RAY DIFFRACTION (2.5 Å)