1 H, 13 C, and 15 N backbone and side chain chemical shift assignments of the SARS-CoV-2 non-structural protein 7
- PDF / 1,078,731 Bytes
- 5 Pages / 595.276 x 790.866 pts Page_size
- 6 Downloads / 199 Views
ARTICLE
1
H, 13C, and 15N backbone and side chain chemical shift assignments of the SARS‑CoV‑2 non‑structural protein 7 Marco Tonelli1 · Chad Rienstra1,2 · Thomas K. Anderson1,3 · Rob Kirchdoerfer1,3 · Katherine Henzler‑Wildman1,2 Received: 3 September 2020 / Accepted: 12 November 2020 © Springer Nature B.V. 2020
Abstract The SARS-CoV-2 genome encodes for approximately 30 proteins. Within the international project covid19-nmr, we distribute the spectroscopic analysis of the viral proteins and RNA. Here, we report NMR chemical shift assignments for the protein nsp7. The 83 amino acid nsp7 protein is an essential cofactor in the RNA-dependent RNA polymerase. The polymerase activity and processivity of nsp12 are greatly enhanced by binding 1 copy of nsp7 and 2 copies of nsp8 to form a 160 kD complex. A separate hexadecameric complex of nsp7 and nsp8 (8 copies of each) forms a large ring-like structure. Thus, nsp7 is an important component of several large protein complexes that are required for replication of the large and complex coronavirus genome. We here report the near-complete NMR backbone and sidechain resonance assignment (1H,13C,15N) of isolated nsp7 from SARS-CoV-2 in solution. Further, we derive the secondary structure and compare it to the previously reported assignments and structure of the SARS-CoV nsp7. Keywords SARS-CoV-2 · Non-structural protein · Solution NMR-spectroscopy · COVID19-NMR
Biological context SARS-CoV-2, the causative agent of COVID-19, emerged late in 2019 to cause a global pandemic. This novel coronavirus is highly related to SARS-CoV that emerged in 2002. While several coronaviruses routinely infect humans causing mild respiratory symptoms (van der Hoek 2007), SARSCoV-2 is capable of causing severe respiratory symptoms or even death (Bchetnia et al. 2020). Coronaviruses are enveloped viruses with large positive-sense RNA genomes. At 30 kb or even greater, some of the genomes of the Nidovirales order, which includes coronaviruses, are among the largest RNA genomes known, requiring a highly processive RNA synthesis process for their replication.
* Katherine Henzler‑Wildman [email protected] 1
National Magnetic Resonance Facility at Madison (NMRFAM), University of Wisconsin at Madison, Madison, WI 53706, USA
2
Department of Biochemistry, University of Wisconsin at Madison, Madison, WI 53706, USA
3
Institute for Molecular Virology, University of Wisconsin at Madison, Madison, WI 53706, USA
Upon entering host cells, the coronavirus RNA genome acts as an mRNA to be translated by host ribosomes to produce the viral polyproteins pp1a and pp1ab. These polyproteins are cleaved to produce a suite of 16 non-structural proteins (nsp) that are responsible for replication and transcription of the viral RNA genome (Snijder et al. 2016). The viral nsp assemble membrane enclosed compartments containing the virus RNA-dependent RNA-polymerase. The minimal polymerase complex required for activity in vitro is composed of nsp7, nsp8 and nsp12. While the polymerase active site is con
Data Loading...
