2R8F

Crystal structure of H225A NSP2 and ATP-gS complex

Summary for 2R8F

• Entry DOI: 10.2210/pdb2r8f/pdb
• Related: 1l9v 2R7C 2R7J 2R7P
• Descriptor: Non-structural RNA-binding protein 35, PHOSPHATE ION, PHOSPHOTHIOPHOSPHORIC ACID-ADENYLATE ESTER, ... (4 entities in total)
• Functional Keywords: rotavirus, ndp kinase, non structural protein, ntpase, rna binding protein, rna-binding
• Biological source: Simian 11 rotavirus (serotype 3 / strain SA11-Ramig)
• Cellular location: Host cytoplasm (Potential): Q03243
• Total number of polymer chains: 1
• Total formula weight: 37169.41

Authors

Kumar, M.,Prasad, B.V.V. (deposition date: 2007-09-10, release date: 2007-10-23, Last modification date: 2023-10-25)

Primary citation

Kumar, M.,Jayaram, H.,Vasquez-Del Carpio, R.,Jiang, X.,Taraporewala, Z.F.,Jacobson, R.H.,Patton, J.T.,Prasad, B.V.V.
Crystallographic and Biochemical Analysis of Rotavirus NSP2 with Nucleotides Reveals a Nucleoside Diphosphate Kinase-Like Activity
J.Virol., 81:12272-12284, 2007

PubMed Abstract: Rotavirus, the major pathogen of infantile gastroenteritis, carries a nonstructural protein, NSP2, essential for viroplasm formation and genome replication/packaging. In addition to RNA-binding and helix-destabilizing properties, NSP2 exhibits nucleoside triphosphatase activity. A conserved histidine (H225) functions as the catalytic residue for this enzymatic activity, and mutation of this residue abrogates genomic double-stranded RNA synthesis without affecting viroplasm formation. To understand the structural basis of the phosphatase activity of NSP2, we performed crystallographic analyses of native NSP2 and a functionally defective H225A mutant in the presence of nucleotides. These studies showed that nucleotides bind inside a cleft between the two domains of NSP2 in a region that exhibits structural similarity to ubiquitous cellular HIT (histidine triad) proteins. Only minor conformational alterations were observed in the cleft upon nucleotide binding and hydrolysis. This hydrolysis involved the formation of a stable phosphohistidine intermediate. These observations, reminiscent of cellular nucleoside diphosphate (NDP) kinases, prompted us to investigate whether NSP2 exhibits phosphoryl-transfer activity. Bioluminometric assay showed that NSP2 exhibits an NDP kinase-like activity that transfers the bound phosphate to NDPs. However, NSP2 is distinct from the highly conserved cellular NDP kinases in both its structure and catalytic mechanism, thus making NSP2 a potential target for antiviral drug design. With structural similarities to HIT proteins, which are not known to exhibit NDP kinase activity, NSP2 represents a unique example among structure-activity relationships. The newly observed phosphoryl-transfer activity of NSP2 may be utilized for homeostasis of nucleotide pools in viroplasms during genome replication.

PubMed: 17804496
DOI: 10.1128/JVI.00984-07

Experimental method

X-RAY DIFFRACTION (2.8 Å)