title: The envelope protein of severe acute respiratory syndrome coronavirus interacts with the non-structural protein 3 and is ubiquitinated.
abstract: To analyze the proteins interacting with the severe acute respiratory syndrome coronavirus (SARS-CoV) envelope (E) protein, a SARS-CoV was engineered including two tags associated to the E protein. Using this virus, complexes of SARS-CoV E and other proteins were purified using a tandem affinity purification system. Several viral and cell proteins including spike, membrane, non-structural protein 3 (nsp3), dynein heavy chain, fatty acid synthase and transmembrane protein 43 bound E protein. In the present work, we focused on the binding of E protein to nsp3 in infected cells and cell-free systems. This interaction was mediated by the N-terminal acidic domain of nsp3. Moreover, nsp3 and E protein colocalized during the infection. It was shown that E protein was ubiquitinated in vitro and in cell culture, suggesting that the interaction between nsp3 and E protein may play a role in the E protein ubiquitination status and therefore on its turnover.
authors.affiliation: Centro Nacional de Biotecnología (CNB), CSIC, Darwin 3, Campus Universidad Autónoma de Madrid, 28049 Madrid, Spain.
authors.forename: Enrique
authors.initials: E
authors.lastname: Alvarez
authors.forename: Marta L
authors.initials: ML
authors.lastname: DeDiego
authors.forename: Jose L
authors.initials: JL
authors.lastname: Nieto-Torres
authors.forename: Jose M
authors.initials: JM
authors.lastname: Jiménez-Guardeño
authors.forename: Laura
authors.initials: L
authors.lastname: Marcos-Villar
authors.forename: Luis
authors.initials: L
authors.lastname: Enjuanes
mesh_terms: D000818:Animals; D002460:Cell Line; D002522:Chlorocebus aethiops; D002846:Chromatography, Affinity; D006801:Humans; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D025941:Protein Interaction Mapping; D011499:Protein Processing, Post-Translational; D012324:RNA-Dependent RNA Polymerase / Q000378:metabolism*; D045473:Severe acute respiratory syndrome-related coronavirus / Q000502:physiology*; D054875:Ubiquitination; D014759:Viral Envelope Proteins / Q000378:metabolism*; D017361:Viral Nonstructural Proteins / Q000378:metabolism*; D000086082:Viroporin Proteins
chemical_list: C501689:E protein, SARS coronavirus; D014759:Viral Envelope Proteins; D017361:Viral Nonstructural Proteins; D000086082:Viroporin Proteins; C510959:Nsp3 protein, SARS-CoV; D012324:RNA-Dependent RNA Polymerase
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