Nuclear Magnetic Resonance Structure Shows that the Severe Acute Respiratory Syndrome Coronavirus-Unique Domain Contains a Macrodomain Fold

• DOI: 10.1128/jvi.01781-08
• Publication Date: 2008-12-04T01:55:23Z
• Authors: Amarnath Chatterjee; Margaret A. Johnson; Pedro Serrano; Bill Pedrini; Jeremiah S. Joseph; Benjamin W. Neuman; Kumar Saikatendu; Michael J. Buchmeier; Peter Kuhn; Kurt Wüthrich
• Keywords: Coronavirus; Protein tertiary structure

The nuclear magnetic resonance (NMR) structure of a central segment the previously annotated severe acute respiratory syndrome (SARS)-unique domain (SUD-M, for "middle SARS-unique domain") in SARS coronavirus (SARS-CoV) nonstructural protein 3 (nsp3) has been determined. SUD-M(513-651) exhibits macrodomain fold containing nsp3 residues 528 to 648, and there is flexibly extended N-terminal tail with 513 527 C-terminal flexible 649 651. As follow-up this initial result, we also solved construct representing only globular 651 [SUD-M(527-651)]. NMR chemical shift perturbation experiments showed that SUD-M(527-651) binds single-stranded poly(A) identified contact area RNA on surface, electrophoretic mobility assays then confirmed SUD-M higher affinity purine bases than pyrimidine bases. In further search clues function, found closest three-dimensional homology another nsp3, ADP-ribose-1"-phosphatase nsp3b, although two proteins share 5% sequence identity homologous regions. shows several helicases nucleoside triphosphate-binding proteins, but it does not contain motifs catalytic these structural homologues. combined results from screening potential substrates structure-based studies now form basis more focused investigations role viral infection.