The SARS-unique domain (SUD) of SARS-CoV and SARS-CoV-2 interacts with human Paip1 to enhance viral RNA translation

EMBO J. 2021 Jun 1;40(11):e102277. doi: 10.15252/embj.2019102277.

Jian Lei ¹ ² ³, Yue Ma-Lauer ⁴ ⁵, Yinze Han ³, Matthias Thoms ⁶, Robert Buschauer ⁶, Joerg Jores ⁷, Volker Thiel ⁸, Roland Beckmann ⁶, Wen Deng ⁹ ¹⁰, Heinrich Leonhardt ⁹, Rolf Hilgenfeld ¹ ² ¹¹, Albrecht von Brunn ⁴ ⁵

Affiliations

1 Institute of Biochemistry, Center for Structural and Cell Biology in Medicine, University of Lübeck, Lübeck, Germany.
2 German Center for Infection Research (DZIF), Hamburg-Lübeck- Borstel-Riems Site, University of Lübeck, Lübeck, Germany.
3 State Key Laboratory of Biotherapy and Cancer Center, National Clinical Research Center for Geriatrics, West China Hospital, Sichuan University, Chengdu, China.
4 Max-von-Pettenkofer Institute, Ludwig-Maximilians-University Munich, Munich, Germany.
5 German Center for Infection Research (DZIF), Munich, Germany.
6 Gene Center Munich, Department of Biochemistry, Ludwig-Maximilians-University Munich, Munich, Germany.
7 Institute of Veterinary Bacteriology, Vetsuisse Faculty, University of Bern, Bern, Switzerland.
8 Institute of Virology and Immunology, University of Bern, Bern, Switzerland.
9 Department of Biology and Center for Integrated Protein Science, Ludwig-Maximilians-University Munich, Planegg-Martinsried, Germany.
10 College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, China.
11 Institute of Molecular Medicine, University of Lübeck, Lübeck, Germany.

PMID: 33876849 DOI: 10.15252/embj.2019102277

Abstract

The ongoing outbreak of severe acute respiratory syndrome (SARS) coronavirus 2 (SARS-CoV-2) demonstrates the continuous threat of emerging coronaviruses (CoVs) to public health. SARS-CoV-2 and SARS-CoV share an otherwise non-conserved part of non-structural protein 3 (Nsp3), therefore named as "SARS-unique domain" (SUD). We previously found a yeast-2-hybrid screen interaction of the SARS-CoV SUD with human poly(A)-binding protein (PABP)-interacting protein 1 (Paip1), a stimulator of protein translation. Here, we validate SARS-CoV SUD:Paip1 interaction by size-exclusion chromatography, split-yellow Fluorescent protein, and co-immunoprecipitation assays, and confirm such interaction also between the corresponding domain of SARS-CoV-2 and Paip1. The three-dimensional structure of the N-terminal domain of SARS-CoV SUD ("macrodomain II", Mac2) in complex with the middle domain of Paip1, determined by X-ray crystallography and small-angle X-ray scattering, provides insights into the structural determinants of the complex formation. In cellulo, SUD enhances synthesis of viral but not host proteins via binding to Paip1 in pBAC-SARS-CoV replicon-transfected cells. We propose a possible mechanism for stimulation of viral translation by the SUD of SARS-CoV and SARS-CoV-2.

Keywords

coronavirus; eukaryotic translation initiation factors; macrodomain; protein synthesis; virus-host interactions.

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