2XHV

HCV-J4 NS5B Polymerase Point Mutant Orthorhombic Crystal Form

2XHV の概要

• エントリーDOI: 10.2210/pdb2xhv/pdb
• 関連するPDBエントリー: 1NB4 1NB6 1NB7 2F55 2XHU 2XHW
• 分子名称: RNA-directed RNA polymerase, SULFATE ION, MAGNESIUM ION, ... (4 entities in total)
• 機能のキーワード: replication, transcription, transferase
• 由来する生物種: Hepatitis C virus genotype 1b (strain HC-J4) (HCV)
• 細胞内の位置: Core protein p21: Host endoplasmic reticulum membrane; Single-pass membrane protein (By similarity). Core protein p19: Virion (By similarity). Envelope glycoprotein E1: Virion membrane; Single-pass type I membrane protein (Potential). Envelope glycoprotein E2: Virion membrane; Single-pass type I membrane protein (Potential). p7: Host endoplasmic reticulum membrane; Multi-pass membrane protein (By similarity). Protease NS2-3: Host endoplasmic reticulum membrane; Multi-pass membrane protein (Potential). Serine protease NS3: Host endoplasmic reticulum membrane; Peripheral membrane protein (By similarity). Non-structural protein 4A: Host endoplasmic reticulum membrane; Single-pass type I membrane protein (Potential). Non-structural protein 4B: Host endoplasmic reticulum membrane; Multi-pass membrane protein (By similarity). Non-structural protein 5A: Host endoplasmic reticulum membrane; Peripheral membrane protein (By similarity). RNA-directed RNA polymerase: Host endoplasmic reticulum membrane; Single-pass type I membrane protein (Potential): O92972
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 131059.92

構造登録者

Harrus, D.,Ahmed-El-Sayed, N.,Simister, P.C.,Miller, S.,Triconnet, M.,Hagedorn, C.H.,Mahias, K.,Rey, F.A.,Astier-Gin, T.,Bressanelli, S. (登録日: 2010-06-21, 公開日: 2010-08-04, 最終更新日: 2024-03-27)

主引用文献

Harrus, D.,Ahmed-El-Sayed, N.,Simister, P.C.,Miller, S.,Triconnet, M.,Hagedorn, C.H.,Mahias, K.,Rey, F.A.,Astier-Gin, T.,Bressanelli, S.
Further Insights Into the Roles of GTP and the C- Terminus of the Hepatitis C Virus Polymerase in the Initiation of RNA Synthesis
J.Biol.Chem., 285:32906-, 2010

PubMed Abstract: The hepatitis C virus (HCV) NS5b protein is an RNA-dependent RNA polymerase essential for replication of the viral RNA genome. In vitro and presumably in vivo, NS5b initiates RNA synthesis by a de novo mechanism. Different structural elements of NS5b have been reported to participate in RNA synthesis, especially a so-called "β-flap" and a C-terminal segment (designated "linker") that connects the catalytic core of NS5b to a transmembrane anchor. High concentrations of GTP have also been shown to stimulate de novo RNA synthesis by HCV NS5b. Here we describe a combined structural and functional analysis of genotype 1 HCV-NS5b of strains H77 (subtype 1a), for which no structure has been previously reported, and J4 (subtype 1b). Our results highlight the linker as directly involved in lifting the first boundary to processive RNA synthesis, the formation of the first dinucleotide primer. The transition from this first dinucleotide primer state to processive RNA synthesis requires removal of the linker and of the β-flap with which it is shown to strongly interact in crystal structures of HCV NS5b. We find that GTP specifically stimulates this transition irrespective of its incorporation in neosynthesized RNA.

PubMed: 20729191
DOI: 10.1074/JBC.M110.151316

実験手法

X-RAY DIFFRACTION (1.9 Å)