2CKW

The 2.3 A resolution structure of the Sapporo virus RNA dependant RNA polymerase.

2CKW の概要

• エントリーDOI: 10.2210/pdb2ckw/pdb
• 分子名称: RNA-DIRECTED RNA POLYMERASE (2 entities in total)
• 機能のキーワード: hydrolase, nucleotide-binding, nucleotidyltransferase, covalent protein-rna linkage, polymerase, rna elongation, transferase activity, mutant, capsid protein, rna replication, structural protein, protease, helicase, transferase, rna-directed rna polymerase, polyprotein, atp-binding, thiol protease
• 由来する生物種: SAPPORO VIRUS
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 56485.71

構造登録者

Fullerton, S.W.B.,Tucker, P.A.,Rohayem, J.,Coutard, B.,Gebhardt, J.,Gorbalenya, A.,Canard, B. (登録日: 2006-04-24, 公開日: 2006-11-29, 最終更新日: 2023-12-13)

主引用文献

Fullerton, S.W.B.,Blaschke, M.,Coutard, B.,Gebhardt, J.,Gorbalenya, A.,Canard, B.,Tucker, P.A.,Rohayem, J.
Structural and Functional Characterization of Sapovirus RNA-Dependent RNA Polymerase.
J.Virol., 81:1858-, 2007

PubMed Abstract: Sapoviruses are one of the major agents of acute gastroenteritis in childhood. They form a tight genetic cluster (genus) in the Caliciviridae family that regroups both animal and human pathogenic strains. No permissive tissue culture has been developed for human sapovirus, limiting its characterization to surrogate systems. We report here on the first extensive characterization of the key enzyme of replication, the RNA-dependent RNA polymerase (RdRp) associated with the 3D(pol)-like protein. Enzymatically active sapovirus 3D(pol) and its defective mutant were expressed in Escherichia coli and purified. The overall structure of the sapovirus 3D(pol) was determined by X-ray crystallography to 2.32-A resolution. It revealed a right hand fold typical for template-dependent polynucleotide polymerases. The carboxyl terminus is located within the active site cleft, as observed in the RdRp of some (norovirus) but not other (lagovirus) caliciviruses. Sapovirus 3D(pol) prefers Mn(2+) over Mg(2+) but may utilize either as a cofactor in vitro. In a synthetic RNA template-dependent reaction, sapovirus 3D(pol) synthesizes a double-stranded RNA or labels the template 3' terminus by terminal transferase activity. Initiation of RNA synthesis occurs de novo on heteropolymeric templates or in a primer-dependent manner on polyadenylated templates. Strikingly, this mode of initiation of RNA synthesis was also described for norovirus, but not for lagovirus, suggesting structural and functional homologies in the RNA-dependent RNA polymerase of human pathogenic caliciviruses. This first experimental evidence makes sapovirus 3D(pol) an attractive target for developing drugs to control calicivirus infection in humans.

PubMed: 17121797
DOI: 10.1128/JVI.01462-06

実験手法

X-RAY DIFFRACTION (2.3 Å)