tetano
Editor, Senior Moderator
EMBO J
. 2021 Apr 20;e102277.
doi: 10.15252/embj.2019102277. Online ahead of print.
The SARS-unique domain (SUD) of SARS-CoV and SARS-CoV-2 interacts with human Paip1 to enhance viral RNA translation
Jian Lei[SUP] 1 2 3 [/SUP], Yue Ma-Lauer[SUP] 4 5 [/SUP], Yinze Han[SUP] 3 [/SUP], Matthias Thoms[SUP] 6 [/SUP], Robert Buschauer[SUP] 6 [/SUP], Joerg Jores[SUP] 7 [/SUP], Volker Thiel[SUP] 8 [/SUP], Roland Beckmann[SUP] 6 [/SUP], Wen Deng[SUP] 9 10 [/SUP], Heinrich Leonhardt[SUP] 9 [/SUP], Rolf Hilgenfeld[SUP] 1 2 11 [/SUP], Albrecht von Brunn[SUP] 4 5 [/SUP]
Affiliations
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
aip1 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.
. 2021 Apr 20;e102277.
doi: 10.15252/embj.2019102277. Online ahead of print.
The SARS-unique domain (SUD) of SARS-CoV and SARS-CoV-2 interacts with human Paip1 to enhance viral RNA translation
Jian Lei[SUP] 1 2 3 [/SUP], Yue Ma-Lauer[SUP] 4 5 [/SUP], Yinze Han[SUP] 3 [/SUP], Matthias Thoms[SUP] 6 [/SUP], Robert Buschauer[SUP] 6 [/SUP], Joerg Jores[SUP] 7 [/SUP], Volker Thiel[SUP] 8 [/SUP], Roland Beckmann[SUP] 6 [/SUP], Wen Deng[SUP] 9 10 [/SUP], Heinrich Leonhardt[SUP] 9 [/SUP], Rolf Hilgenfeld[SUP] 1 2 11 [/SUP], Albrecht von Brunn[SUP] 4 5 [/SUP]
Affiliations
- 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
aip1 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.