4WTG

CRYSTAL STRUCTURE OF HCV NS5B GENOTYPE 2A JFH-1 ISOLATE WITH S15G E86Q E87Q C223H V321I MUTATIONS AND DELTA8 BETA HAIRPIN LOOP DELETION IN COMPLEX WITH SOFOSBUVIR DIPHOSPHATE GS-607596, MN2+ AND SYMMETRICAL PRIMER TEMPLATE 5'-CAAAAUUU

Summary for 4WTG

• Entry DOI: 10.2210/pdb4wtg/pdb
• Descriptor: RNA PRIMER TEMPLATE CAAAAUUU, RNA-directed RNA polymerase, MANGANESE (II) ION, ... (6 entities in total)
• Functional Keywords: hcv, viral, ns5b, rdrp, resistance mutation, template, primer, primed initiation, delta8 beta hairpin loop deletion, transferase-rna complex, transferase/rna
• Biological source: Hepatitis C virus JFH-1; More
• 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: 3
• Total formula weight: 69366.75

Authors

Edwards, T.E.,Appleby, T.C. (deposition date: 2014-10-30, release date: 2015-02-11, Last modification date: 2023-09-27)

Primary citation

Appleby, T.C.,Perry, J.K.,Murakami, E.,Barauskas, O.,Feng, J.,Cho, A.,Fox, D.,Wetmore, D.R.,McGrath, M.E.,Ray, A.S.,Sofia, M.J.,Swaminathan, S.,Edwards, T.E.
Structural basis for RNA replication by the hepatitis C virus polymerase.
Science, 347:771-775, 2015

PubMed Abstract: Nucleotide analog inhibitors have shown clinical success in the treatment of hepatitis C virus (HCV) infection, despite an incomplete mechanistic understanding of NS5B, the viral RNA-dependent RNA polymerase. Here we study the details of HCV RNA replication by determining crystal structures of stalled polymerase ternary complexes with enzymes, RNA templates, RNA primers, incoming nucleotides, and catalytic metal ions during both primed initiation and elongation of RNA synthesis. Our analysis revealed that highly conserved active-site residues in NS5B position the primer for in-line attack on the incoming nucleotide. A β loop and a C-terminal membrane-anchoring linker occlude the active-site cavity in the apo state, retract in the primed initiation assembly to enforce replication of the HCV genome from the 3' terminus, and vacate the active-site cavity during elongation. We investigated the incorporation of nucleotide analog inhibitors, including the clinically active metabolite formed by sofosbuvir, to elucidate key molecular interactions in the active site.

PubMed: 25678663
DOI: 10.1126/science.1259210

Experimental method

X-RAY DIFFRACTION (2.9 Å)