Limits...
A Dimerization-Dependent Mechanism Drives the Endoribonuclease Function of Porcine Reproductive and Respiratory Syndrome Virus nsp11.

Shi Y, Li Y, Lei Y, Ye G, Shen Z, Sun L, Luo R, Wang D, Fu ZF, Xiao S, Peng G - J. Virol. (2016)

Bottom Line: The PRRSV nsp11 endoribonuclease plays a vital role in arterivirus replication, but its precise roles and mechanisms of action are poorly understood.Structural and biochemical experiments demonstrated that nsp11 exists mainly as a dimer in solution and that nsp11 may be fully active as a dimer.Mutagenesis and structural analysis revealed NendoU active site residues, which are conserved throughout the order Nidovirales(families Arteriviridae and Coronaviridae) and the major determinants of dimerization (Ser74 and Phe76) in Arteriviridae Importantly, these findings may provide a new structural basis for antiviral drug development.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan, China College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, China.

Show MeSH

Related in: MedlinePlus

The structural comparison of the N-terminal region of PRRSV nsp11 and the middle region of coronavirus nsp15. (A and B) The structure of PRRSV nsp11 (subunit A, yellow) superimposed onto the structures of SARS-CoV nsp15 (PDB code 2H85, magenta) and MHV nsp15 (PDB code 2GTH, magenta). The structure of the N-terminal region (Gly1 to Gly106) from PRRSV nsp11 superimposed with SARS-CoV nsp15 (Asn62-Ser197) and MHV nsp15 (Ser62-Leu228) is enlarged in panels A and B (the cartoon transparency was set at 60%). The dimerization site determinants Ser74 and Phe76 (corresponding to Val162/Leu156 and Leu167/Val161 in SARS-CoV nsp15 and MHV nsp15, respectively) are labeled with a ball-and-stick (yellow, PRRSV nsp11; magenta, SARS-CoV nsp15 and MHV nsp15) representation. The SARS-CoV nsp15 domains are colored and marked as described for Fig. 3C.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4836315&req=5

Figure 4: The structural comparison of the N-terminal region of PRRSV nsp11 and the middle region of coronavirus nsp15. (A and B) The structure of PRRSV nsp11 (subunit A, yellow) superimposed onto the structures of SARS-CoV nsp15 (PDB code 2H85, magenta) and MHV nsp15 (PDB code 2GTH, magenta). The structure of the N-terminal region (Gly1 to Gly106) from PRRSV nsp11 superimposed with SARS-CoV nsp15 (Asn62-Ser197) and MHV nsp15 (Ser62-Leu228) is enlarged in panels A and B (the cartoon transparency was set at 60%). The dimerization site determinants Ser74 and Phe76 (corresponding to Val162/Leu156 and Leu167/Val161 in SARS-CoV nsp15 and MHV nsp15, respectively) are labeled with a ball-and-stick (yellow, PRRSV nsp11; magenta, SARS-CoV nsp15 and MHV nsp15) representation. The SARS-CoV nsp15 domains are colored and marked as described for Fig. 3C.

Mentions: Our crystal structure indicates that nsp11 assembles into dimers, which is different from coronavirus nsp15 (19, 21). The monomer structure of SARS-CoV nsp15 includes three domains, the N-terminal domain (NTD), the middle domain, and the catalytic domain (23) (Fig. 3C); the NTD is critical for hexamerization and interactions with the middle domain and the catalytic domain of an adjacent monomer (19, 23). However, the NTD structure (approximately 61 N-terminal residues) of coronavirus nsp15 is missing in nsp11, and the NTD of nsp11 superimposes onto the middle domain of coronavirus nsp15 (Fig. 4). Moreover, the major determinants of dimerization (Ser74 and Phe76) are significantly different from the key residues involved in the oligomerization of SARS-CoV nsp15 (Fig. 4; Fig. 5), which indicates why the active form of nsp11 is a dimer rather than a hexamer.


A Dimerization-Dependent Mechanism Drives the Endoribonuclease Function of Porcine Reproductive and Respiratory Syndrome Virus nsp11.

Shi Y, Li Y, Lei Y, Ye G, Shen Z, Sun L, Luo R, Wang D, Fu ZF, Xiao S, Peng G - J. Virol. (2016)

The structural comparison of the N-terminal region of PRRSV nsp11 and the middle region of coronavirus nsp15. (A and B) The structure of PRRSV nsp11 (subunit A, yellow) superimposed onto the structures of SARS-CoV nsp15 (PDB code 2H85, magenta) and MHV nsp15 (PDB code 2GTH, magenta). The structure of the N-terminal region (Gly1 to Gly106) from PRRSV nsp11 superimposed with SARS-CoV nsp15 (Asn62-Ser197) and MHV nsp15 (Ser62-Leu228) is enlarged in panels A and B (the cartoon transparency was set at 60%). The dimerization site determinants Ser74 and Phe76 (corresponding to Val162/Leu156 and Leu167/Val161 in SARS-CoV nsp15 and MHV nsp15, respectively) are labeled with a ball-and-stick (yellow, PRRSV nsp11; magenta, SARS-CoV nsp15 and MHV nsp15) representation. The SARS-CoV nsp15 domains are colored and marked as described for Fig. 3C.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4836315&req=5

Figure 4: The structural comparison of the N-terminal region of PRRSV nsp11 and the middle region of coronavirus nsp15. (A and B) The structure of PRRSV nsp11 (subunit A, yellow) superimposed onto the structures of SARS-CoV nsp15 (PDB code 2H85, magenta) and MHV nsp15 (PDB code 2GTH, magenta). The structure of the N-terminal region (Gly1 to Gly106) from PRRSV nsp11 superimposed with SARS-CoV nsp15 (Asn62-Ser197) and MHV nsp15 (Ser62-Leu228) is enlarged in panels A and B (the cartoon transparency was set at 60%). The dimerization site determinants Ser74 and Phe76 (corresponding to Val162/Leu156 and Leu167/Val161 in SARS-CoV nsp15 and MHV nsp15, respectively) are labeled with a ball-and-stick (yellow, PRRSV nsp11; magenta, SARS-CoV nsp15 and MHV nsp15) representation. The SARS-CoV nsp15 domains are colored and marked as described for Fig. 3C.
Mentions: Our crystal structure indicates that nsp11 assembles into dimers, which is different from coronavirus nsp15 (19, 21). The monomer structure of SARS-CoV nsp15 includes three domains, the N-terminal domain (NTD), the middle domain, and the catalytic domain (23) (Fig. 3C); the NTD is critical for hexamerization and interactions with the middle domain and the catalytic domain of an adjacent monomer (19, 23). However, the NTD structure (approximately 61 N-terminal residues) of coronavirus nsp15 is missing in nsp11, and the NTD of nsp11 superimposes onto the middle domain of coronavirus nsp15 (Fig. 4). Moreover, the major determinants of dimerization (Ser74 and Phe76) are significantly different from the key residues involved in the oligomerization of SARS-CoV nsp15 (Fig. 4; Fig. 5), which indicates why the active form of nsp11 is a dimer rather than a hexamer.

Bottom Line: The PRRSV nsp11 endoribonuclease plays a vital role in arterivirus replication, but its precise roles and mechanisms of action are poorly understood.Structural and biochemical experiments demonstrated that nsp11 exists mainly as a dimer in solution and that nsp11 may be fully active as a dimer.Mutagenesis and structural analysis revealed NendoU active site residues, which are conserved throughout the order Nidovirales(families Arteriviridae and Coronaviridae) and the major determinants of dimerization (Ser74 and Phe76) in Arteriviridae Importantly, these findings may provide a new structural basis for antiviral drug development.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan, China College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, China.

Show MeSH
Related in: MedlinePlus