4WTL

CRYSTAL STRUCTURE OF HCV NS5B GENOTYPE 2A JFH-1 ISOLATE WITH S15G E86Q E87Q C223H V321I MUTATIONS IN COMPLEX WITH RNA TEMPLATE 5'-UACC, RNA PRIMER 5'-PGG, MN2+, AND UDP

4WTL の概要

• エントリーDOI: 10.2210/pdb4wtl/pdb
• 分子名称: RNA TEMPLATE UACC, RNA PRIMER GG, RNA-directed RNA polymerase, ... (9 entities in total)
• 機能のキーワード: hcv, viral, ns5b, rdrp, resistance mutation, template, primer, primed initiation, transferase-rna complex, transferase/rna
• 由来する生物種: Hepatitis C virus JFH-1; 詳細
• 細胞内の位置: 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
• タンパク質・核酸の鎖数: 3
• 化学式量合計: 67763.77

構造登録者

Edwards, T.E.,Appleby, T.C. (登録日: 2014-10-30, 公開日: 2015-02-11, 最終更新日: 2023-09-27)

主引用文献

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

実験手法

X-RAY DIFFRACTION (2 Å)