Regulation of epithelial sodium channel activity by SARS-CoV-1 and SARS-CoV-2 proteins.
Regulation of epithelial sodium channel activity by SARS-CoV-1 and SARS-CoV-2 proteins.
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DOI：
10.1016/j.bpj.2021.06.005
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发表时间：
2021-07-20
影响因子：
3.4
通讯作者：
Lester HA
中科院分区：
文献类型：
--
作者：
Grant SN;Lester HA
严重急性呼吸综合征（SARS）冠状病毒（CoV）2（SARS-CoV-2），导致冠状病毒疾病2019，编码几种蛋白质，其作用知之甚少。我们测试了它们直接形成质膜离子通道或改变两种哺乳动物质膜离子通道（上皮钠通道（ENaC）和α3β4烟碱乙酰胆碱受体）功能的能力。在注射了mRNA的非洲爪蟾卵母细胞中，9种SARS-CoV-2蛋白或2种SARS-CoV-1蛋白都没有产生电导，几种组合的共注射也没有。ORF 8、刺突（S）和包膜（E）蛋白的免疫印迹显示，这些蛋白以适当的分子量表达。在与ENaC共表达的实验中，三种测试的SARS蛋白（SARS-CoV-1 E、SARS-CoV-2 E和SARS-CoV-2 S）显著降低ENaC电流。SARS-CoV-1 S蛋白适度降低ENaC电流。与α3β4烟碱型乙酰胆碱受体共表达E蛋白而非S蛋白显著降低乙酰胆碱诱导的电流。如果在ENaC mRNA后24小时注射SARS-CoV蛋白mRNA，则ENaC抑制不会发生，这表明SARS-CoV蛋白影响通道蛋白功能表达的早期步骤。与SARS-CoV-2 S蛋白诱导的ENaC抑制涉及对可用蛋白酶的竞争的假设一致，突变SARS-CoV-2 S蛋白中的弗林蛋白酶切割位点部分地减轻了ENaC电流的抑制。扩展了先前关于SARS蛋白通过蛋白激酶C（PKC）激活影响ENaC电流的建议，通过佛波醇12-肉豆蔻酸酯13-乙酸酯激活PKC降低ENaC和α3β4活性。佛波醇12-肉豆蔻酸酯13-乙酸酯的应用使膜电容降低了约5%，推测是通过增加内吞作用，但这种降低比SARS蛋白对电导的影响小得多。此外，在PKCα和PKCβ抑制剂Gö-6976中孵育卵母细胞，并没有改变E或S蛋白诱导的通道抑制。我们的结论是，SARS-CoV-1和SARS-CoV-2蛋白改变人类质膜通道的功能，通过不完全理解的机制。这些相互作用可能在冠状病毒2019的病理生理学中发挥作用。
Severe acute respiratory syndrome (SARS) coronavirus (CoV) 2 (SARS-CoV-2), which causes the coronavirus disease 2019, encodes several proteins whose roles are poorly understood. We tested their ability either to directly form plasma membrane ion channels or to change functions of two mammalian plasma membrane ion channels, the epithelial sodium channel (ENaC) and the α3β4 nicotinic acetylcholine receptor. In mRNA-injected Xenopus oocytes, none of nine SARS-CoV-2 proteins or two SARS-CoV-1 proteins produced conductances, nor did co-injection of several combinations. Immunoblots for ORF8, spike (S), and envelope (E) proteins revealed that the proteins are expressed at appropriate molecular weights. In experiments on coexpression with ENaC, three tested SARS proteins (SARS-CoV-1 E, SARS-CoV-2 E, and SARS-CoV-2 S) markedly decrease ENaC currents. SARS-CoV-1 S protein decreases ENaC currents modestly. Coexpressing the E proteins but not the S proteins with α3β4 nicotinic acetylcholine receptors significantly reduces acetylcholine-induced currents. ENaC inhibition does not occur if the SARS-CoV protein mRNAs are injected 24 h after the ENaC mRNAs, suggesting that SARS-CoV proteins affect early step(s) in functional expression of channel proteins. Consistent with the hypothesis that the SARS-CoV-2 S protein-induced ENaC inhibition involves competition for available protease, mutating the furin cleavage site in SARS-CoV-2 S protein partially relieves inhibition of ENaC currents. Extending previous suggestions that SARS proteins affect ENaC currents via protein kinase C (PKC) activation, PKC activation via phorbol 12-myristate 13-acetate decreases ENaC and α3β4 activity. Phorbol 12-myristate 13-acetate application reduced membrane capacitance ∼5%, presumably via increased endocytosis, but this decrease is much smaller than the SARS proteins’ effects on conductances. Also, incubating oocytes in Gö-6976, a PKCα and PKCβ inhibitor, did not alter E or S protein-induced channel inhibition. We conclude that SARS-CoV-1 and SARS-CoV-2 proteins alter the function of human plasma membrane channels, via incompletely understood mechanisms. These interactions may play a role in the coronavirus 2019 pathophysiology.
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影响因子：
8.8
作者：
Mulay A;Konda B;Garcia G Jr;Yao C;Beil S;Villalba JM;Koziol C;Sen C;Purkayastha A;Kolls JK;Pociask DA;Pessina P;de Aja JS;Garcia-de-Alba C;Kim CF;Gomperts B;Arumugaswami V;Stripp BR
通讯作者：
Stripp BR
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
影响因子：
64.8
作者：
Delorey TM;Ziegler CGK;Heimberg G;Normand R;Yang Y;Segerstolpe Å;Abbondanza D;Fleming SJ;Subramanian A;Montoro DT;Jagadeesh KA;Dey KK;Sen P;Slyper M;Pita-Juárez YH;Phillips D;Biermann J;Bloom-Ackermann Z;Barkas N;Ganna A;Gomez J;Melms JC;Katsyv I;Normandin E;Naderi P;Popov YV;Raju SS;Niezen S;Tsai LT;Siddle KJ;Sud M;Tran VM;Vellarikkal SK;Wang Y;Amir-Zilberstein L;Atri DS;Beechem J;Brook OR;Chen J;Divakar P;Dorceus P;Engreitz JM;Essene A;Fitzgerald DM;Fropf R;Gazal S;Gould J;Grzyb J;Harvey T;Hecht J;Hether T;Jané-Valbuena J;Leney-Greene M;Ma H;McCabe C;McLoughlin DE;Miller EM;Muus C;Niemi M;Padera R;Pan L;Pant D;Pe'er C;Pfiffner-Borges J;Pinto CJ;Plaisted J;Reeves J;Ross M;Rudy M;Rueckert EH;Siciliano M;Sturm A;Todres E;Waghray A;Warren S;Zhang S;Zollinger DR;Cosimi L;Gupta RM;Hacohen N;Hibshoosh H;Hide W;Price AL;Rajagopal J;Tata PR;Riedel S;Szabo G;Tickle TL;Ellinor PT;Hung D;Sabeti PC;Novak R;Rogers R;Ingber DE;Jiang ZG;Juric D;Babadi M;Farhi SL;Izar B;Stone JR;Vlachos IS;Solomon IH;Ashenberg O;Porter CBM;Li B;Shalek AK;Villani AC;Rozenblatt-Rosen O;Regev A
通讯作者：
Regev A
影响因子：
5.5
作者：
Cabrera-Garcia, David;Bekdash, Ramsey;Harrison, Neil L.
通讯作者：
Harrison, Neil L.
影响因子：
6.4
作者：
Castaño-Rodriguez C;Honrubia JM;Gutiérrez-Álvarez J;DeDiego ML;Nieto-Torres JL;Jimenez-Guardeño JM;Regla-Nava JA;Fernandez-Delgado R;Verdia-Báguena C;Queralt-Martín M;Kochan G;Perlman S;Aguilella VM;Sola I;Enjuanes L
通讯作者：
Enjuanes L