The crystal structure of porcine reproductive and respiratory syndrome virus nonstructural protein Nsp1beta reveals a novel metal-dependent nuclease.
Porcine reproductive and respiratory syndrome virus (PRRSV), a member of the Arteriviridae family of Nidovirales, is the causative agent of porcine reproductive and respiratory syndrome, which results in enormous economic losses in the swine industry. As the second protein encoded by the PRRSV genome, nsp1beta cleaves itself from the downstream nsp2 protein via a C-terminal papain-like cysteine protease (PCP) domain. Although nsp1beta is known to be involved in virulence, its precise role in the process of viral infection remains unclear. In this work, we describe the homodimeric crystal structure of PRRSV nsp1beta in its natural, self-processed form. We show that the architecture of its N-terminal domain (NTD) adopts a fold closely resembling that of several known nucleases and has intrinsic nuclease activity that is strongly activated by manganese ions in vitro. Key features, however, distinguish nsp1beta from characterized nucleases, including the C-terminal PCP domain (which is responsible for the self-release of nsp1beta from nsp2), a linker domain (LKD) that connects the NTD and the PCP domain, and a C-terminal extension (CTE) that binds to and is stabilized by the putative substrate binding site of the PCPbeta domain. Combined with the reported nuclear localization of this protein, these results shed light on the self-processing mode and precise biological function of nsp1beta and thus offer a multitarget template for future drug discovery.
Xue
Fei
F
Structural Biology Laboratory, Tsinghua University, Beijing 100084, China.
Sun
Yuna
Y
Yan
Liming
L
Zhao
Cong
C
Chen
Ji
J
Bartlam
Mark
M
Li
Xuemei
X
Lou
Zhiyong
Z
Rao
Zihe
Z
D000595:Amino Acid Sequence; D003067:Coenzymes / Q000494:pharmacology*; D018360:Crystallography, X-Ray; D004279:DNA, Viral / Q000378:metabolism; D003851:Deoxyribonucleases / Q000737:chemistry* / Q000378:metabolism*; D019281:Dimerization; D008345:Manganese / Q000494:pharmacology*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D019316:Porcine respiratory and reproductive syndrome virus / Q000737:chemistry*; D020836:Protein Structure, Quaternary; D016415:Sequence Alignment; D017361:Viral Nonstructural Proteins / Q000737:chemistry* / Q000378:metabolism*
D003067:Coenzymes; D004279:DNA, Viral; D017361:Viral Nonstructural Proteins; D008345:Manganese; D003851:Deoxyribonucleases
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