SND1 binds SARS-CoV-2 negative-sense RNA and promotes viral RNA synthesis through NSP9.
SND1 binds SARS-CoV-2 negative-sense RNA and promotes viral RNA synthesis through NSP9.
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DOI：
10.1016/j.cell.2023.09.002
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发表时间：
2023-10-26
期刊：
影响因子：
64.5
通讯作者：
Munschauer, Mathias
中科院分区：
文献类型：
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作者：
Schmidt, Nora;Ganskih, Sabina;Wei, Yuanjie;Gabel, Alexander;Zielinski, Sebastian;Keshishian, Hasmik;Lareau, Caleb A.;Zimmermann, Liv;Makroczyova, Jana;Pearce, Cadence;Krey, Karsten;Hennig, Thomas;Stegmaier, Sebastian;Moyon, Lambert;Horlacher, Marc;Werner, Simone;Aydin, Jens;Olguin-Nava, Marco;Potabattula, Ramya;Kibe, Anuja;Doelken, Lars;Smyth, Redmond P.;Caliskan, Neva;Marsico, Annalisa;Krempl, Christine;Bodem, Jochen;Pichlmair, Andreas;Carr, Steven A.;Chlanda, Petr;Erhard, Florian;Munschauer, Mathias
病毒RNA生物合成的调节是生产性SARS-CoV-2感染的基础。为了表征参与这一过程的宿主RNA结合蛋白（RBP），我们通过生物化学方法鉴定了与基因组和亚基因组SARS-CoV-2 RNA结合的蛋白质。我们发现宿主蛋白SND 1与负义病毒RNA的5′端结合，是SARS-CoV-2 RNA合成所必需的。SND 1缺失的细胞形成较小的复制细胞器，并显示出减弱的病毒生长动力学。我们发现NSP 9是病毒RBP和SND 1直接相互作用的伴侣，与感染过程中产生的正义和负义RNA的5′端共价连接。这些连接发生在复制-转录起始位点，与NSP 9引发病毒RNA合成一致。在机制上，SND 1重塑NSP 9占据并改变NSP 9与病毒RNA中起始核苷酸的共价键。我们的研究结果表明NSP 9参与了SARS-CoV-2 RNA合成的启动，并揭示了一种细胞蛋白在协调病毒RNA产生中的未知作用。SND 1是感染早期新生SARS-CoV-2 RNA合成所必需的SND 1直接与NSP 9相互作用，两种蛋白质都与负义病毒RNA结合。NSP 9在起始位点与病毒RNA共价连接，表明蛋白质引发SND 1调节NSP 9与正义和负义病毒RNA的共价连接。蛋白质相互作用揭示了宿主蛋白SND 1结合负义SARS-CoV-2 RNA并通过募集NSP 9促进病毒RNA合成，NSP 9可能作为RNA产生的蛋白质引物。
Regulation of viral RNA biogenesis is fundamental to productive SARS-CoV-2 infection. To characterize host RNA-binding proteins (RBPs) involved in this process, we biochemically identified proteins bound to genomic and subgenomic SARS-CoV-2 RNAs. We find that the host protein SND1 binds the 5′ end of negative-sense viral RNA and is required for SARS-CoV-2 RNA synthesis. SND1-depleted cells form smaller replication organelles and display diminished virus growth kinetics. We discover that NSP9, a viral RBP and direct SND1 interaction partner, is covalently linked to the 5′ ends of positive- and negative-sense RNAs produced during infection. These linkages occur at replication-transcription initiation sites, consistent with NSP9 priming viral RNA synthesis. Mechanistically, SND1 remodels NSP9 occupancy and alters the covalent linkage of NSP9 to initiating nucleotides in viral RNA. Our findings implicate NSP9 in the initiation of SARS-CoV-2 RNA synthesis and unravel an unsuspected role of a cellular protein in orchestrating viral RNA production. SND1 is required for nascent SARS-CoV-2 RNA synthesis early during infection SND1 directly interacts with NSP9 and both proteins bind negative-sense viral RNA NSP9 is covalently linked to viral RNA at initiation sites, indicating protein priming SND1 modulates the covalent linkage of NSP9 to positive- and negative-sense viral RNA Mapping of subgenome-resolved SARS-CoV-2 RNA-protein interactions reveals that the host protein SND1 binds negative-sense SARS-CoV-2 RNA and promotes viral RNA synthesis by recruiting NSP9, which likely serves as a protein primer for RNA production.
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影响因子：
64.5
作者：
Hafner M;Landthaler M;Burger L;Khorshid M;Hausser J;Berninger P;Rothballer A;Ascano M Jr;Jungkamp AC;Munschauer M;Ulrich A;Wardle GS;Dewell S;Zavolan M;Tuschl T
通讯作者：
Tuschl T
影响因子：
3.7
作者：
Galán C;Sola I;Nogales A;Thomas B;Akoulitchev A;Enjuanes L;Almazán F
通讯作者：
Almazán F
DOI：
10.1007/s00018-020-03745-y
发表时间：
2021-04
期刊：
Cellular and molecular life sciences : CMLS
影响因子：
--
作者：
Eymieux S;Rouillé Y;Terrier O;Seron K;Blanchard E;Rosa-Calatrava M;Dubuisson J;Belouzard S;Roingeard P
通讯作者：
Roingeard P
影响因子：
5.6
作者：
Friberg, Anders;Corsini, Lorenzo;Sattler, Michael
通讯作者：
Sattler, Michael
影响因子：
30.8
作者：
Basak, Anindita;Munschauer, Mathias;Sankaran, Vijay G.
通讯作者：
Sankaran, Vijay G.