title: Murine norovirus protein NS1/2 aspartate to glutamate mutation, sufficient for persistence, reorients side chain of surface exposed tryptophan within a novel structured domain.
abstract: Compact viral genomes such as those found in noroviruses, which cause significant enteric disease in humans, often encode only a few proteins, but affect a wide range of processes in their hosts and ensure efficient propagation of the virus. Both human and mouse noroviruses (MNVs) persistently replicate and are shed in stool, a highly effective strategy for spreading between hosts. For MNV, the presence of a glutamate rather than an aspartate at position 94 of the NS1/2 protein was previously shown to be essential for persistent replication and shedding. Here, we analyze these critical sequences of NS1/2 at the structural level. Using solution nuclear magnetic resonance methods, we determined folded NS1/2 domain structures from a nonpersistent murine norovirus strain CW3, a persistent strain CR6, and a persistent mutant strain CW3(D94E). We found an unstructured PEST-like domain followed by a novel folded domain in the N-terminus of NS1/2. All three forms of the domain are stable and monomeric in solution. Residue 94, critical for determining persistence, is located in a reverse turn following an α-helix in the folded domain. The longer side chain of glutamate, but not aspartate, allows interaction with the indole group of the nearby tryptophan, reshaping the surface of the domain. The discrimination between glutamyl and aspartyl residue is imposed by the stable tertiary conformation. These structural requirements correlate with the in vivo function of NS1/2 in persistence, a key element of norovirus biology and infection.
authors.affiliation: NMR Accelerator, San Diego, California.
authors.forename: Brendan N
authors.initials: BN
authors.lastname: Borin
authors.forename: Wei
authors.initials: W
authors.lastname: Tang
authors.forename: Timothy J
authors.initials: TJ
authors.lastname: Nice
authors.forename: Broc T
authors.initials: BT
authors.lastname: McCune
authors.forename: Herbert W
authors.initials: HW
authors.lastname: Virgin
authors.forename: Andrzej M
authors.initials: AM
authors.lastname: Krezel
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids / Q000737:chemistry / Q000235:genetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics*; D029322:Norovirus / Q000737:chemistry / Q000235:genetics; D011487:Protein Conformation; D017361:Viral Nonstructural Proteins / Q000737:chemistry / Q000235:genetics
chemical_list: D000596:Amino Acids; D017361:Viral Nonstructural Proteins
keywords: MNV; PEST motif; helix-strand-helix; polyprotein; protein NMR; side chain rotamers