Oligomeric State of β-Coronavirus Non-Structural Protein 10 Stimulators Studied by Small Angle X-ray Scattering

Wolfgang Knecht, S. Z. Fisher, Jiaqi Lou, Céleste Sele, Shumeng Ma, Anna A. Rasmussen, Nikos Pinotsis, Frank Kozielski

    Research output: Contribution to journalArticlepeer-review

    Abstract

    The β-coronavirus family, encompassing Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), Severe Acute Respiratory Syndrome Coronavirus (SARS), and Middle East Respiratory Syndrome Coronavirus (MERS), has triggered pandemics within the last two decades. With the possibility of future pandemics, studying the coronavirus family members is necessary to improve knowledge and treatment. These viruses possess 16 non-structural proteins, many of which play crucial roles in viral replication and in other vital functions. One such vital protein is non-structural protein 10 (nsp10), acting as a pivotal stimulator of nsp14 and nsp16, thereby influencing RNA proofreading and viral RNA cap formation. Studying nsp10 of pathogenic coronaviruses is central to unraveling its multifunctional roles. Our study involves the biochemical and biophysical characterisation of full-length nsp10 from MERS, SARS and SARS-CoV-2. To elucidate their oligomeric state, we employed a combination of Multi-detection Size exclusion chromatography (Multi-detection SEC) with multi-angle static light scattering (MALS) and small angle X-ray scattering (SAXS) techniques. Our findings reveal that full-length nsp10s primarily exist as monomers in solution, while truncated versions tend to oligomerise. SAXS experiments reveal a globular shape for nsp10, a trait conserved in all three coronaviruses, although MERS nsp10, diverges most from SARS and SARS-CoV-2 nsp10s. In summary, unbound nsp10 proteins from SARS, MERS, and SARS-CoV-2 exhibit a globular and predominantly monomeric state in solution.
    Original languageEnglish
    Article number13649
    Number of pages14
    JournalInternational Journal of Molecular Sciences
    Volume24
    Issue number17
    DOIs
    Publication statusPublished - 2023 Sept 4

    Subject classification (UKÄ)

    • Biochemistry and Molecular Biology

    Free keywords

    • SARS-CoV-2
    • COVID-19
    • non-structural proteins
    • nsp10
    • SAXS
    • oligomeric state
    • conformational changes

    Fingerprint

    Dive into the research topics of 'Oligomeric State of β-Coronavirus Non-Structural Protein 10 Stimulators Studied by Small Angle X-ray Scattering'. Together they form a unique fingerprint.

    Cite this