Processing of the SARS-CoV pp1a/ab nsp7–10 region
Identifieur interne : 000366 ( 2020/Analysis ); précédent : 000365; suivant : 000367Processing of the SARS-CoV pp1a/ab nsp7–10 region
Auteurs : Boris Krichel ; Sven Falke ; Rolf Hilgenfeld ; Lars Redecke ; Charlotte UetrechtSource :
- Biochemical Journal [ 0264-6021 ] ; 2020.
Abstract
Severe acute respiratory syndrome coronavirus is the causative agent of a respiratory disease with a high case fatality rate. During the formation of the coronaviral replication/transcription complex, essential steps include processing of the conserved polyprotein nsp7–10 region by the main protease Mpro and subsequent complex formation of the released nsp's. Here, we analyzed processing of the coronavirus nsp7–10 region using native mass spectrometry showing consumption of substrate, rise and fall of intermediate products and complexation. Importantly, there is a clear order of cleavage efficiencies, which is influenced by the polyprotein tertiary structure. Furthermore, the predominant product is an nsp7+8(2 : 2) hetero-tetramer with nsp8 scaffold. In conclusion, native MS, opposed to other methods, can expose the processing dynamics of viral polyproteins and the landscape of protein interactions in one set of experiments. Thereby, new insights into protein interactions, essential for generation of viral progeny, were provided, with relevance for development of antivirals.
Url:
DOI: 10.1042/BCJ20200029
PubMed: 32083638
PubMed Central: 7078746
Affiliations:
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During the formation of the coronaviral replication/transcription complex, essential steps include processing of the conserved polyprotein nsp7–10 region by the main protease M<sup>pro</sup> and subsequent complex formation of the released nsp's. Here, we analyzed processing of the coronavirus nsp7–10 region using native mass spectrometry showing consumption of substrate, rise and fall of intermediate products and complexation. Importantly, there is a clear order of cleavage efficiencies, which is influenced by the polyprotein tertiary structure. Furthermore, the predominant product is an nsp7+8(2 : 2) hetero-tetramer with nsp8 scaffold. In conclusion, native MS, opposed to other methods, can expose the processing dynamics of viral polyproteins and the landscape of protein interactions in one set of experiments. 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Moll</name></author><author><name sortKey="Weimar, T" uniqKey="Weimar T">T. Weimar</name></author><author><name sortKey="Mesters, J R" uniqKey="Mesters J">J.R. Mesters</name></author><author><name sortKey="Hilgenfeld, R" uniqKey="Hilgenfeld R">R. Hilgenfeld</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Liu, F" uniqKey="Liu F">F. Liu</name></author><author><name sortKey="Rijkers, D T S" uniqKey="Rijkers D">D.T.S. Rijkers</name></author><author><name sortKey="Post, H" uniqKey="Post H">H. Post</name></author><author><name sortKey="Heck, A J R" uniqKey="Heck A">A.J.R. Heck</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Specht, H" uniqKey="Specht H">H. Specht</name></author><author><name sortKey="Emmott, E" uniqKey="Emmott E">E. Emmott</name></author><author><name sortKey="Petelski, A" uniqKey="Petelski A">A. Petelski</name></author><author><name sortKey="Huffman, R G" uniqKey="Huffman R">R.G. Huffman</name></author><author><name sortKey="Perlman, D H" uniqKey="Perlman D">D.H. Perlman</name></author><author><name sortKey="Serra, M" uniqKey="Serra M">M. Serra</name></author></analytic></biblStruct></listBibl></div1></back></TEI><affiliations><list></list><tree><noCountry><name sortKey="Falke, Sven" sort="Falke, Sven" uniqKey="Falke S" first="Sven" last="Falke">Sven Falke</name><name sortKey="Hilgenfeld, Rolf" sort="Hilgenfeld, Rolf" uniqKey="Hilgenfeld R" first="Rolf" last="Hilgenfeld">Rolf Hilgenfeld</name><name sortKey="Krichel, Boris" sort="Krichel, Boris" uniqKey="Krichel B" first="Boris" last="Krichel">Boris Krichel</name><name sortKey="Redecke, Lars" sort="Redecke, Lars" uniqKey="Redecke L" first="Lars" last="Redecke">Lars Redecke</name><name sortKey="Uetrecht, Charlotte" sort="Uetrecht, Charlotte" uniqKey="Uetrecht C" first="Charlotte" last="Uetrecht">Charlotte Uetrecht</name></noCountry></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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|area= SrasV1
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|texte= Processing of the SARS-CoV pp1a/ab nsp7–10 region
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