Structural basis for the interaction of SARS-CoV-2 virulence factor nsp1 with DNA polymerase α-primase.
Structural basis for the interaction of SARS-CoV-2 virulence factor nsp1 with DNA polymerase α-primase.
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DOI：
10.1002/pro.4220
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发表时间：
2022-03
期刊：
影响因子：
--
通讯作者：
Pellegrini L
中科院分区：
文献类型：
--
作者：
Kilkenny ML;Veale CE;Guppy A;Hardwick SW;Chirgadze DY;Rzechorzek NJ;Maman JD;Pellegrini L
驱动严重急性呼吸道综合征冠状病毒2型（SARS-CoV-2）感染的分子机制-2019冠状病毒病（COVID-19）的病原体-目前正在密切关注，以了解病毒如何运作，并揭示预防或缓解疾病的方法。最近对病毒和宿主蛋白之间相互作用的蛋白质组学筛选已经确定了SARS-CoV-2靶向的人类蛋白质。DNA聚合酶α（Pol α）-引发酶复合物或引发体-负责在基因组复制期间启动DNA合成-被鉴定为非结构蛋白1（nsp 1）的靶标，nsp 1是SARS-CoV-2感染中的主要毒力因子。在这里，我们验证发表的报告的相互作用的nsp 1与primosome证明直接结合纯化的重组成分，并提供了一个生化表征它们的相互作用。此外，我们通过阐明与原体结合的nsp 1的冷冻电子显微镜结构，为相互作用提供了结构基础。我们的研究结果为毒力因子nsp 1靶向Pol α的报道提供了生化证据，并表明SARS-CoV-2干扰了Pol α在病毒感染期间免疫应答中的假定作用。PDB代码：7 OPL; EMDB代码：13020、13021
The molecular mechanisms that drive the infection by the severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2)—the causative agent of coronavirus disease 2019 (COVID‐19)—are under intense current scrutiny to understand how the virus operates and to uncover ways in which the disease can be prevented or alleviated. Recent proteomic screens of the interactions between viral and host proteins have identified the human proteins targeted by SARS‐CoV‐2. The DNA polymerase α (Pol α)–primase complex or primosome—responsible for initiating DNA synthesis during genomic duplication—was identified as a target of nonstructural protein 1 (nsp1), a major virulence factor in the SARS‐CoV‐2 infection. Here, we validate the published reports of the interaction of nsp1 with the primosome by demonstrating direct binding with purified recombinant components and providing a biochemical characterization of their interaction. Furthermore, we provide a structural basis for the interaction by elucidating the cryo‐electron microscopy structure of nsp1 bound to the primosome. Our findings provide biochemical evidence for the reported targeting of Pol α by the virulence factor nsp1 and suggest that SARS‐CoV‐2 interferes with Pol α's putative role in the immune response during the viral infection. PDB Code(s): 7OPL; EMDB codes: 13020, 13021
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影响因子：
16
作者：
Kolinjivadi AM;Sannino V;De Antoni A;Zadorozhny K;Kilkenny M;Técher H;Baldi G;Shen R;Ciccia A;Pellegrini L;Krejci L;Costanzo V
通讯作者：
Costanzo V
DOI：
10.1126/science.abe9403
发表时间：
2020-12-04
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Gordon DE;Hiatt J;Bouhaddou M;Rezelj VV;Ulferts S;Braberg H;Jureka AS;Obernier K;Guo JZ;Batra J;Kaake RM;Weckstein AR;Owens TW;Gupta M;Pourmal S;Titus EW;Cakir M;Soucheray M;McGregor M;Cakir Z;Jang G;O'Meara MJ;Tummino TA;Zhang Z;Foussard H;Rojc A;Zhou Y;Kuchenov D;Hüttenhain R;Xu J;Eckhardt M;Swaney DL;Fabius JM;Ummadi M;Tutuncuoglu B;Rathore U;Modak M;Haas P;Haas KM;Naing ZZC;Pulido EH;Shi Y;Barrio-Hernandez I;Memon D;Petsalaki E;Dunham A;Marrero MC;Burke D;Koh C;Vallet T;Silvas JA;Azumaya CM;Billesbølle C;Brilot AF;Campbell MG;Diallo A;Dickinson MS;Diwanji D;Herrera N;Hoppe N;Kratochvil HT;Liu Y;Merz GE;Moritz M;Nguyen HC;Nowotny C;Puchades C;Rizo AN;Schulze-Gahmen U;Smith AM;Sun M;Young ID;Zhao J;Asarnow D;Biel J;Bowen A;Braxton JR;Chen J;Chio CM;Chio US;Deshpande I;Doan L;Faust B;Flores S;Jin M;Kim K;Lam VL;Li F;Li J;Li YL;Li Y;Liu X;Lo M;Lopez KE;Melo AA;Moss FR 3rd;Nguyen P;Paulino J;Pawar KI;Peters JK;Pospiech TH Jr;Safari M;Sangwan S;Schaefer K;Thomas PV;Thwin AC;Trenker R;Tse E;Tsui TKM;Wang F;Whitis N;Yu Z;Zhang K;Zhang Y;Zhou F;Saltzberg D;QCRG Structural Biology Consortium;Hodder AJ;Shun-Shion AS;Williams DM;White KM;Rosales R;Kehrer T;Miorin L;Moreno E;Patel AH;Rihn S;Khalid MM;Vallejo-Gracia A;Fozouni P;Simoneau CR;Roth TL;Wu D;Karim MA;Ghoussaini M;Dunham I;Berardi F;Weigang S;Chazal M;Park J;Logue J;McGrath M;Weston S;Haupt R;Hastie CJ;Elliott M;Brown F;Burness KA;Reid E;Dorward M;Johnson C;Wilkinson SG;Geyer A;Giesel DM;Baillie C;Raggett S;Leech H;Toth R;Goodman N;Keough KC;Lind AL;Zoonomia Consortium;Klesh RJ;Hemphill KR;Carlson-Stevermer J;Oki J;Holden K;Maures T;Pollard KS;Sali A;Agard DA;Cheng Y;Fraser JS;Frost A;Jura N;Kortemme T;Manglik A;Southworth DR;Stroud RM;Alessi DR;Davies P;Frieman MB;Ideker T;Abate C;Jouvenet N;Kochs G;Shoichet B;Ott M;Palmarini M;Shokat KM;García-Sastre A;Rassen JA;Grosse R;Rosenberg OS;Verba KA;Basler CF;Vignuzzi M;Peden AA;Beltrao P;Krogan NJ
通讯作者：
Krogan NJ
DOI：
10.1107/s0907444904019158
发表时间：
2004-12-01
影响因子：
2.2
作者：
Emsley, P;Cowtan, K
通讯作者：
Cowtan, K
DOI：
10.1073/pnas.0603144103
发表时间：
2006-08-22
影响因子：
11.1
作者：
Kamitani, Wataru;Narayanan, Krishna;Makino, Shinji
通讯作者：
Makino, Shinji
DOI：
10.1074/jbc.m112.368951
发表时间：
2012-07-06
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Kilkenny ML;De Piccoli G;Perera RL;Labib K;Pellegrini L
通讯作者：
Pellegrini L