TY - JOUR

T1 - Structural insights into the activity regulation of full-length non-structural protein 1 from SARS-CoV-2

AU - Wang, Ying

AU - Kirkpatrick, John

AU - Lage, Susanne zur

AU - Carlomagno, Teresa

N1 - Funding Information: We thank Dr Andrea Graziadei (TU Berlin) for the kind gift of the Nsp1 expression plasmid and Prof. Dr. Harald Schwalbe (Goethe University Frankfurt, BMRZ) and Dr. Julia Weigand (Technical University Darmstadt) for the kind gift of the DNA template plasmids of the 5′ UTR region of the SARS-CoV-2. This work was funded by the Deutsche Forschungsgemeinschaft through grant CA294/16-1 to T.C. We thank Dr Andrea Graziadei (TU Berlin) for the kind gift of the Nsp1 expression plasmid and Prof. Dr. Harald Schwalbe (Goethe University Frankfurt, BMRZ) and Dr. Julia Weigand (Technical University Darmstadt) for the kind gift of the DNA template plasmids of the 5′ UTR region of the SARS-CoV-2. This work was funded by the Deutsche Forschungsgemeinschaft through grant CA294/16-1 to T.C. Y.W. and J.K. performed research, analyzed data, and wrote the manuscript; S.z.L. performed research; and T.C. designed and supervised the project, analyzed data, and wrote the manuscript. The authors declare no competing interests. We support inclusive, diverse, and equitable conduct of research.

PY - 2023/2/2

Y1 - 2023/2/2

N2 - Non-structural protein 1 (Nsp1) of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a major virulence factor and thus an attractive drug target. The last 33 amino acids of Nsp1 have been shown to bind within the mRNA entry tunnel of the 40S ribosomal subunit, shutting off host gene expression. Here, we report the solution-state structure of full-length Nsp1, which features an α/β fold formed by a six-stranded, capped β-barrel-like globular domain (N-terminal domain [NTD]), flanked by short N-terminal and long C-terminal flexible tails. The NTD has been found to be critical for 40S-mediated viral mRNA recognition and promotion of viral gene expression. We find that in free Nsp1, the NTD mRNA-binding surface is occluded by interactions with the acidic C-terminal tail, suggesting a mechanism of activity regulation based on the interplay between the folded NTD and the disordered C-terminal region. These results are relevant for drug-design efforts targeting Nsp1.

AB - Non-structural protein 1 (Nsp1) of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a major virulence factor and thus an attractive drug target. The last 33 amino acids of Nsp1 have been shown to bind within the mRNA entry tunnel of the 40S ribosomal subunit, shutting off host gene expression. Here, we report the solution-state structure of full-length Nsp1, which features an α/β fold formed by a six-stranded, capped β-barrel-like globular domain (N-terminal domain [NTD]), flanked by short N-terminal and long C-terminal flexible tails. The NTD has been found to be critical for 40S-mediated viral mRNA recognition and promotion of viral gene expression. We find that in free Nsp1, the NTD mRNA-binding surface is occluded by interactions with the acidic C-terminal tail, suggesting a mechanism of activity regulation based on the interplay between the folded NTD and the disordered C-terminal region. These results are relevant for drug-design efforts targeting Nsp1.

KW - disordered protein domains

KW - drug targets

KW - NMR spectroscopy

KW - non-structural proteins

KW - Nsp1

KW - SARS-CoV-2

UR - http://www.scopus.com/inward/record.url?scp=85147029540&partnerID=8YFLogxK

U2 - 10.1016/j.str.2022.12.006

DO - 10.1016/j.str.2022.12.006

M3 - Article

C2 - 36610391

AN - SCOPUS:85147029540

VL - 31

SP - 128-137.e5

JO - STRUCTURE

JF - STRUCTURE

SN - 0969-2126

IS - 2

ER -