2RHB

Crystal structure of Nsp15-H234A mutant- Hexamer in asymmetric unit

2RHB の概要

• エントリーDOI: 10.2210/pdb2rhb/pdb
• 関連するPDBエントリー: 2gth 2h85
• 分子名称: Uridylate-specific endoribonuclease (2 entities in total)
• 機能のキーワード: endoribonuclease, sars, hexamer, rna, nsp, viral protein
• 由来する生物種: SARS coronavirus
• 細胞内の位置: Non-structural protein 3: Host membrane; Multi-pass membrane protein (Potential). Non-structural protein 4: Host membrane; Multi-pass membrane protein (Potential). Non-structural protein 6: Host membrane; Multi-pass membrane protein (Potential). Non-structural protein 7: Host cytoplasm, host perinuclear region (By similarity). Non-structural protein 8: Host cytoplasm, host perinuclear region (By similarity). Non-structural protein 9: Host cytoplasm, host perinuclear region (By similarity). Non-structural protein 10: Host cytoplasm, host perinuclear region (By similarity). Helicase: Host endoplasmic reticulum-Golgi intermediate compartment (Potential). Uridylate-specific endoribonuclease: Host cytoplasm, host perinuclear region (By similarity): P59641
• タンパク質・核酸の鎖数: 6
• 化学式量合計: 236554.76

構造登録者

Palaninathan, S.,Bhardwaj, K.,Alcantara, J.M.O.,Guarino, L.,Yi, L.L.,Kao, C.C.,Sacchettini, J. (登録日: 2007-10-08, 公開日: 2007-11-27, 最終更新日: 2023-08-30)

主引用文献

Bhardwaj, K.,Palaninathan, S.,Alcantara, J.M.,Yi, L.L.,Guarino, L.,Sacchettini, J.C.,Kao, C.C.
Structural and functional analyses of the severe acute respiratory syndrome coronavirus endoribonuclease Nsp15.
J.Biol.Chem., 283:3655-3664, 2008

PubMed Abstract: The severe acute respiratory syndrome (SARS) coronavirus encodes several RNA-processing enzymes that are unusual for RNA viruses, including Nsp15 (nonstructural protein 15), a hexameric endoribonuclease that preferentially cleaves 3' of uridines. We solved the structure of a catalytically inactive mutant version of Nsp15, which was crystallized as a hexamer. The structure contains unreported flexibility in the active site of each subunit. Substitutions in the active site residues serine 293 and proline 343 allowed Nsp15 to cleave at cytidylate, whereas mutation of leucine 345 rendered Nsp15 able to cleave at purines as well as pyrimidines. Mutations that targeted the residues involved in subunit interactions generally resulted in the formation of catalytically inactive monomers. The RNA-binding residues were mapped by a method linking reversible cross-linking, RNA affinity purification, and peptide fingerprinting. Alanine substitution of several residues in the RNA-contacting portion of Nsp15 did not affect hexamer formation but decreased the affinity of RNA binding and reduced endonuclease activity. This suggests a model for Nsp15 hexamer interaction with RNA.

PubMed: 18045871
DOI: 10.1074/jbc.M708375200

実験手法

X-RAY DIFFRACTION (2.8 Å)