high-throughput-screening-for-broad-spectrum-chemical-inhibitors-rna

High-throughput Screening for Broad-spectrum Chemical Inhibitors of RNA Viruses

RNA viruses are responsible for major human diseases such as flu, bronchitis, dengue, Hepatitis C or measles. They also represent an emerging threat ...

Institut Pasteur, CNRS UMR3569

Dengue disease is an increasing global health problem that threatens one-third of the world's population. Despite decades of efforts, no licensed vaccine against dengue is available. With the aim to develop an affordable vaccine that could be used in young populations living in tropical areas, we evaluated a new strategy based on the expression of a minimal dengue antigen by a vector derived from pediatric live-attenuated Schwarz measles vaccine (MV). As a proof-of-concept, we inserted into the MV vector a sequence encoding a minimal combined dengue antigen composed of the envelope domain III (EDIII) fused to the ectodomain of the membrane protein (ectoM) from DV serotype-1. Immunization of mice susceptible to MV resulted in a long-term production of DV1 serotype-specific neutralizing antibodies. The presence of ectoM was critical to the immunogenicity of inserted EDIII. The adjuvant capacity of ectoM correlated with its ability to promote the maturation of dendritic cells and the secretion of proinflammatory and antiviral cytokines and chemokines involved in adaptive immunity. The protective efficacy of this vaccine should be studied in non-human primates. A combined measles-dengue vaccine might provide a one-shot approach to immunize children against both diseases where they co-exist.

Pediatric measles vaccine expressing a dengue antigen induces durable serotype-specific neutralizing antibodies to dengue virus.

&#x57FA;&#x5B54;&#x80AF;&#x4E9A;&#x70ED;&#x75C5;&#x6BD2; (CHIKV) &#x662F;&#x5BFC;&#x81F4;&#x4EBA;&#x7C7B;&#x4E2D;&#x7684; polyarthralgia&#x3001; &#x53D1;&#x70E7;&#x3001; &#x808C;&#x8089;&#x75DB;&#x3001; &#x5934;&#x75DB;&#x7684;&#x7279;&#x70B9;&#x662F;&#x4E00;&#x79CD;&#x6025;&#x6027;&#x611F;&#x67D3;&#x7684;&#x868A;&#x5B50;&#x4F20;&#x64AD;&#x7532;&#x3002;&#x81EA; 2005 &#x5E74;&#x4EE5;&#x6765;&#x8FD9;&#x79CD;&#x75C5;&#x6BD2;&#x4E00;&#x76F4;&#x8D1F;&#x8D23;&#x7A7A;&#x524D;&#x89C4;&#x6A21;&#x7684;&#x66B4;&#x53D1;&#x6D41;&#x884C;&#x3002;&#x901A;&#x8FC7;&#x4E0E;&#x5176;&#x4ED6;&#x7532;&#x7684;&#x7C7B;&#x6BD4;&#xFF0C;&#x4EBA;&#x4EEC;&#x8BA4;&#x4E3A;&#x7EC6;&#x80DE;&#x86CB;&#x767D;&#x9176;&#x662F;&#x80FD;&#x591F;&#x5904;&#x7406;&#x75C5;&#x6BD2;&#x524D;&#x4F53;&#x86CB;&#x767D; E3E2 &#x751F;&#x4EA7;&#x4F5C;&#x4E3A;&#x9ED1;&#x8179; E1 &#x76F8;&#x5173;&#x8054;&#x7684;&#x53D7;&#x4F53;&#x7ED3;&#x5408; E2 &#x86CB;&#x767D;&#x3002;&#x4E0D;&#x7A33;&#x5B9A;&#x7684;&#x66B4;&#x9732;&#x4E8E;&#x4F4E; pH &#x7684;&#x9ED1;&#x8179;&#x5141;&#x8BB8;&#x75C5;&#x6BD2;&#x878D;&#x5408;&#x548C;&#x611F;&#x67D3;&#x3002;&#x6211;&#x4EEC;&#x663E;&#x793A;&#x9664;&#x7EC6;&#x83CC;&#x6027; convertases &#x5927;&#x578B;&#x9762;&#x677F;&#xFF0C;&#x819C;&#x6027;&#x5F17;&#x6797;&#x86CB;&#x767D;&#x9176;&#xFF0C;&#x4F46; PC5B &#x53EF;&#x4EE5;&#x5904;&#x7406; E3E2 &#x4ECE;&#x975E;&#x6D32; CHIKV &#x83CC;&#x682A;&#x5728; HRQRR(64) / ST &#x7F51;&#x7AD9;&#xFF0C;&#x800C;&#x4E9A;&#x6D32;&#x8D77;&#x6E90; CHIKV &#x5E94;&#x53D8;&#x5206;&#x89E3;&#x5728; RRQRR(64) / SI &#x7684;&#x819C;&#x6027;&#x548C;&#x53EF;&#x6EB6;&#x6027;&#x5F17;&#x6797;&#x86CB;&#x767D;&#x9176;&#x3001; PC5A&#x3001; PC5B&#x3001; PACE4&#xFF0C;&#x800C;&#x4E0D;&#x662F;&#x7531; PC7 &#x6216;&#x6ED1;&#x96EA;-1&#x3002;&#x4F7F;&#x7528;&#x8367;&#x5149;&#x6A21;&#x578B;&#x80BD;&#x548C;&#x91CD;&#x7EC4; convertases&#xFF0C;&#x6211;&#x4EEC;&#x89C2;&#x5BDF;&#x5230;&#x4E9A;&#x6D32;&#x682A; E3E2 &#x6A21;&#x578B;&#x80BD;&#x7684;&#x5F17;&#x6797;&#x86CB;&#x767D;&#x9176;&#x548C; PC5A &#x7684;&#x5207;&#x5272;&#x662F;&#x6548;&#x7387;&#x6700;&#x9AD8;&#x3002;&#x8FD9;&#x79CD;&#x5206;&#x88C2;&#x72B6;&#x51B5;&#x4E5F;&#x6709;&#x4EBA;&#x5728; CHIKV &#x611F;&#x67D3;&#x7EC6;&#x80DE;&#x4E2D;&#xFF0C;&#x800C;&#x53EF;&#x4EE5;&#x88AB;&#x5F17;&#x6797;&#x86CB;&#x767D;&#x9176;&#x6291;&#x5236;&#x5242;&#x5355;-RVKR-&#x6C2F;&#x7532;&#x57FA;&#x916E;&#x963B;&#x6B62;&#x3002;&#x8FD9;&#x6291;&#x5236;&#x4E86;&#x76F8;&#x6BD4;&#x6C2F;&#x55B9;&#xFF0C;&#x5176;&#x80FD;&#x591F;&#x6291;&#x5236; CHIKV &#x8513;&#x5EF6;&#x6210;&#x808C;&#x7EC6;&#x80DE;&#x57F9;&#x517B;&#x7269;&#xFF0C;&#x5728;&#x524D;&#x9762;&#x6240;&#x8FF0;&#x7684;&#x8FD9;&#x79CD;&#x75C5;&#x6BD2;&#x7684;&#x5929;&#x7136;&#x76EE;&#x6807;&#x4F5C;&#x4E3A;&#x5355;&#x5143;&#x683C;&#x7C7B;&#x578B;&#x3002;&#x6211;&#x4EEC;&#x7684;&#x7814;&#x7A76;&#x7ED3;&#x679C;&#x8868;&#x660E;&#x5F17;&#x6797;&#x86CB;&#x767D;&#x9176;&#x6837;&#x86CB;&#x767D;&#x9176; CHIKV &#x9897;&#x7C92;&#x52A0;&#x5DE5;&#x4E2D;&#x7684;&#x4F5C;&#x7528;&#x548C;&#x6307;&#x51FA;&#x65B0;&#x7684;&#x529E;&#x6CD5;&#x6765;&#x6291;&#x5236;&#x6B64;&#x7C7B;&#x611F;&#x67D3;&#x3002;

&#x5F17;&#x6797;&#x86CB;&#x767D;&#x9176;&#x6291;&#x5236;&#x5242;&#x7EC6;&#x80DE;&#x5728;&#x4F53;&#x5916;&#x57F9;&#x517B;&#x7684;&#x4EBA;&#x4F53;&#x808C;&#x8089;&#x7684;&#x57FA;&#x5B54;&#x80AF;&#x4E9A;&#x75C5;&#x75C5;&#x6BD2;&#x611F;&#x67D3;&#x7684;&#x6291;&#x5236;&#x4F5C;&#x7528;&#xFF1A; E2 &#x8868;&#x9762;&#x7CD6;&#x86CB;&#x767D;&#x7684;&#x6210;&#x719F;&#x7684;&#x635F;&#x4F24;&#x3002;

Chikungunya virus (CHIKV) is a mosquito-transmitted Alphavirus that causes in humans an acute infection characterized by polyarthralgia, fever, myalgia, and headache. Since 2005 this virus has been responsible for an epidemic outbreak of unprecedented magnitude. By analogy with other alphaviruses, it is thought that cellular proteases are able to process the viral precursor protein E3E2 to produce the receptor-binding E2 protein that associates as a heterodimer with E1. Destabilization of the heterodimer by exposure to low pH allows viral fusion and infection. We show that among a large panel of proprotein convertases, membranous furin but also PC5B can process E3E2 from African CHIKV strains at the HRQRR(64) / ST site, whereas a CHIKV strain of Asian origin is cleaved at RRQRR(64) / SI by membranous and soluble furin, PC5A, PC5B, and PACE4 but not by PC7 or SKI-1. Using fluorogenic model peptides and recombinant convertases, we observed that the Asian strain E3E2 model peptide is cleaved most efficiently by furin and PC5A. This cleavage was also observed in CHIKV-infected cells and could be blocked by furin inhibitor decanoyl-RVKR-chloromethyl ketone. This inhibitor was compared with chloroquine for its ability to inhibit CHIKV spreading in myoblast cell cultures, a cell-type previously described as a natural target of this virus. Our results demonstrate the role of furin-like proteases in the processing of CHIKV particles and point out new approaches to inhibit this infection.

Inhibition of Chikungunya virus infection in cultured human muscle cells by furin inhibitors: impairment of the maturation of the E2 surface glycoprotein.

Le virus Chikungunya (CHIKV) est un Alphavirus transmises moustique qui provoque chez l&#39;homme une infection aigu&euml; caract&eacute;ris&eacute;e par des maux de t&ecirc;te, fi&egrave;vre, myalgie et polyarthralgie. Depuis 2005, ce virus est responsable d&#39;une &eacute;pid&eacute;mie d&#39;une ampleur sans pr&eacute;c&eacute;dent. Par analogie avec d&#39;autres alphavirus, on croit que les prot&eacute;ases cellulaires sont en mesure de traiter la prot&eacute;ine virale pr&eacute;curseur E3E2 de produire la prot&eacute;ine E2-liaison aux r&eacute;cepteurs qui associe comme un h&eacute;t&eacute;rodim&egrave;re avec E1. Maladies infectieuses et virale fusion permet de d&eacute;stabilisation de l&#39;h&eacute;t&eacute;rodim&egrave;re par l&#39;exposition &agrave; un pH faible. Nous montrons que parmi un large panel des proprot&eacute;ines convertases, membraneuse furine, mais aussi PC5B peut traiter des E3E2 de souches africaines CHIKV &agrave; l&#39;HRQRR(64) / ST du site, alors qu&#39;une souche CHIKV d&#39;origine asiatique est cliv&eacute;e &agrave; RRQRR(64) / SI par furine membranaire et soluble, PC5A, PC5B et PACE4 mais pas par le PC7 ou SKI-1. &Agrave; l&#39;aide de peptides mod&egrave;les fluorog&eacute;niques et convertases recombinante, nous avons observ&eacute; que le peptide de mod&egrave;le de E3E2 de souche asiatique est cliv&eacute; plus efficacement par la furine et PC5A. Ce clivage a &eacute;galement &eacute;t&eacute; observ&eacute; dans les cellules infect&eacute;es par le CHIKV et peut &ecirc;tre bloqu&eacute; par decanoyl-RVKR-chlorom&eacute;thyl c&eacute;tone de furine inhibiteur. Cet inhibiteur a &eacute;t&eacute; compar&eacute; &agrave; la chloroquine pour sa capacit&eacute; &agrave; inhiber la CHIKV r&eacute;pand dans les cultures de cellules myoblastes, un type de cellule pr&eacute;c&eacute;demment d&eacute;crit comme une cible naturelle de ce virus. Nos r&eacute;sultats d&eacute;montrent le r&ocirc;le des prot&eacute;ases dans le traitement des particules CHIKV furine et signaler de nouvelles approches pour inhiber cette infection.

L&#39;inhibition de l&#39;infection par le virus Chikungunya dans le muscle humain cultiv&eacute; des cellules par les inhibiteurs de la furine : alt&eacute;ration de la maturation de la glycoprot&eacute;ine de surface E2.

Chikungunya-Virus (CHIKV) ist ein M&uuml;cken &uuml;bertragene Alphavirus, die beim Menschen eine akute Infektion, charakterisiert durch Polyarthralgia, Fieber, Myalgie und Kopfschmerzen verursacht. Seit 2005 ist dieses Virus verantwortlich f&uuml;r epidemischen Ausbruch der beispiellosen Gr&ouml;&szlig;enordnung. Analog zu anderen Alphaviruses sind der Ansicht, dass zellul&auml;re Proteasen sind verarbeiten die virale Vorl&auml;uferprotein E3E2 Rezeptor-Bindung E2 Proteins zu produzieren, die als ein Heterodimer E1 zuordnet. Destabilisierung der Heterodimer durch Aussetzung zu den niedrigen pH-Wert kann virale Fusion und Infektion. Wir zeigen, dass unter einer gro&szlig;en Gruppe von proprotein Convertases, membran&ouml;se Furin verarbeiten, sondern auch PC5B E3E2 aus afrikanischen CHIKV St&auml;mmen an der HRQRR(64) k&ouml;nnen / ST Platz, w&auml;hrend ein CHIKV Stamm von asiatischer Herkunft am RRQRR(64) zerspaltet wird / SI von membran&ouml;se und l&ouml;slichen Furin, PC5A, PC5B und PACE4 aber nicht von PC7 oder SKI-1. Mit Fluorogenic Modell Peptide und rekombinante Convertases, beobachteten wir, dass das asiatischen Stamm E3E2 Modell Peptid am effizientesten durch Furin und PC5A zerspaltet wird. Diese Spaltung war auch bei CHIKV-infizierten Zellen beobachtet und Furin Inhibitor Decanoyl-RVKR-Chlormethyl Keton geblockt werden konnte. Dieser Inhibitor verglich mit Chloroquin f&uuml;r seine F&auml;higkeit, CHIKV breitet sich in Myoblast Zellkulturen, eine Art von Zelle zuvor beschrieben als nat&uuml;rliches Ziel dieses Virus hemmen. Unsere Ergebnisse zeigen die Rolle der Furin-&auml;hnlichen Proteasen bei der Verarbeitung von CHIKV Partikel und darauf hin neue Ans&auml;tze f&uuml;r diese Infektion zu hemmen.

Hemmung der Chikungunya-Virus-Infektion in kultivierten menschlichen Muskel-Zellen durch Furin-Inhibitoren: Wertminderung von der Reifung von der Oberfl&auml;che E2-Glykoprotein.

Chikungunya virus (CHIKV) &egrave; un Alphavirus trasmesso da zanzare che provoca nell&#39;uomo un&#39;infezione acuta caratterizzata da polyarthralgia, febbre, mialgia e cefalea. Dal 2005 questo virus &egrave; stato responsabile di un focolaio epidemico di ampiezza senza precedenti. Per analogia con altri alphaviruses, si &egrave; pensato che le proteasi cellulari sono in grado di elaborare la proteina virale precursore E3E2 di produrre la proteina E2 receptor-binding che associa come un eterodimero con E1. Destabilizzazione dell&#39;eterodimero da esposizione a pH basso consente di infezione e fusione virale. Mostriamo che tra un grande pannello di proprotein convertasi, furin membranosa, ma anche PC5B pu&ograve; elaborare E3E2 da ceppi africani CHIKV presso il HRQRR(64) / ST sito, considerando che un ceppo CHIKV di origine asiatica &egrave; fenduto a RRQRR(64) / SI furin membranoso e solubile, PC5A, PC5B e PACE4 ma non PC7 o sci-1. Usando peptidi modello fluorogenic e ricombinante convertasi, abbiamo osservato che il peptide di ceppo asiatico E3E2 modello viene scisso in modo pi&ugrave; efficiente di furin e PC5A. Questa scissione &egrave; stato anche osservato in cellule infettate da CHIKV e potrebbe essere bloccato da furin chetone di inibitore clorometil-decanoyl-RVKR. Questo inibitore &egrave; stato confrontato con clorochina per la sua capacit&agrave; di inibire il CHIKV diffondendo in colture di cellule del myoblast, un tipo di cellula, precedentemente descritto come una destinazione naturale di questo virus. I nostri risultati dimostrano il ruolo delle proteasi furin-come nell&#39;elaborazione delle particelle CHIKV e segnalare nuovi approcci per inibire questa infezione.

Inibizione di infezione da virus Chikungunya nel muscolo umano colto le cellule da furin inibitori: alterazione della maturazione della glicoproteina di superficie E2.

&#x30C1;&#x30AF;&#x30F3;&#x30B0;&#x30CB;&#x30A2; &#x30A6;&#x30A4;&#x30EB;&#x30B9; (CHIKV) &#x306F;&#x3001;&#x4EBA;&#x9593;&#x306E; polyarthralgia&#x3001;&#x767A;&#x71B1;&#x3001;&#x7B4B;&#x8089;&#x75DB;&#x3001;&#x982D;&#x75DB;&#x306B;&#x3088;&#x3063;&#x3066;&#x7279;&#x5FB4;&#x4ED8;&#x3051;&#x3089;&#x308C;&#x308B;&#x6025;&#x6027;&#x611F;&#x67D3;&#x75C7;&#x3092;&#x5F15;&#x304D;&#x8D77;&#x3053;&#x3059;&#x868A;&#x8EE2;&#x9001;&#x30A2;&#x30EB;&#x30D5;&#x30A1;&#x30A6;&#x30A4;&#x30EB;&#x30B9;&#x3067;&#x3059;&#x3002;2005 &#x5E74;&#x4EE5;&#x6765;&#x3053;&#x306E;&#x30A6;&#x30A4;&#x30EB;&#x30B9;&#x306F;&#x3001;&#x524D;&#x4F8B;&#x306E;&#x306A;&#x3044;&#x5927;&#x304D;&#x3055;&#x306E;&#x6D41;&#x884C;&#x767A;&#x751F;&#x3092;&#x62C5;&#x5F53;&#x3057;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;&#x305D;&#x306E;&#x4ED6;&#x30A2;&#x30EB;&#x30D5;&#x30A1;&#x30A6;&#x30A4;&#x30EB;&#x30B9;&#x5C5E;&#x3068;&#x306E;&#x30A2;&#x30CA;&#x30ED;&#x30B8;&#x30FC;&#x3067;&#x306F;&#x3001;&#x7D30;&#x80DE;&#x306E;&#x30D7;&#x30ED;&#x30C6;&#x30A2;&#x30FC;&#x30BC;&#x304C;&#x30A6;&#x30A4;&#x30EB;&#x30B9;&#x306E;&#x524D;&#x99C6;&#x7269;&#x8CEA;&#x86CB;&#x767D;&#x8CEA;&#x3068;&#x3057;&#x3066;&#x30D8;&#x30C6;&#x30ED;&#x30C0;&#x30A4;&#x30DE;&#x30FC; E1 &#x3068;&#x95A2;&#x9023;&#x4ED8;&#x3051;&#x307E;&#x3059;&#x53D7;&#x5BB9;&#x4F53;&#x7D50;&#x5408; E2 &#x86CB;&#x767D;&#x8CEA;&#x3092;&#x4F5C;&#x308A;&#x51FA;&#x3059; E3E2 &#x3092;&#x51E6;&#x7406;&#x3067;&#x304D;&#x3066;&#x3044;&#x308B;&#x3053;&#x3068;&#x304C;&#x8003;&#x3048;&#x3089;&#x308C;&#x307E;&#x3059;&#x3002;&#x4F4E; pH &#x3078;&#x306E;&#x66B4;&#x9732;&#x306B;&#x3088;&#x3063;&#x3066;&#x30D8;&#x30C6;&#x30ED;&#x30C0;&#x30A4;&#x30DE;&#x30FC;&#x306E;&#x4E0D;&#x5B89;&#x5B9A;&#x5316;&#x3067;&#x306F;&#x3001;&#x611F;&#x67D3;&#x3068;&#x30A6;&#x30A4;&#x30EB;&#x30B9;&#x878D;&#x5408;&#x3092;&#x3053;&#x3068;&#x304C;&#x3067;&#x304D;&#x307E;&#x3059;&#x3002;&#x79C1;&#x9054;&#x306F;&#x30D7;&#x30ED;&#x30BF;&#x30F3;&#x30D1;&#x30AF;&#x304A;&#x306E;&#x5927;&#x578B;&#x30D1;&#x30CD;&#x30EB;&#x306E;&#x9593;&#x3067;&#x306F;&#x3001;&#x819C;&#x8CEA;&#x306E;&#x98A8;&#x9234;&#x304C; PC5B E3E2&#x3001;HRQRR(64) &#x3067;&#x306E;&#x30A2;&#x30D5;&#x30EA;&#x30AB;&#x306E; CHIKV &#x682A;&#x304B;&#x3089;&#x51E6;&#x7406;&#x3067;&#x304D;&#x308B;&#x3053;&#x3068;&#x3092;&#x793A;&#x3059;/ST &#x306E;&#x30B5;&#x30A4;&#x30C8;&#x306F;&#x3001;CHIKV &#x3072;&#x305A;&#x307F;&#x30A2;&#x30B8;&#x30A2;&#x8D77;&#x6E90;&#x306E; RRQRR(64) &#x3067;&#x5207;&#x65AD;&#x3055;&#x308C;&#x308B;&#x306B;&#x5BFE;&#x3057;/SI &#x819C;&#x3068;&#x53EF;&#x6EB6;&#x6027;&#x98A8;&#x9234;&#x3001;PC5A&#x3001;PC5B&#x3001;&#x304A;&#x3088;&#x3073; PACE4 &#x304C; PC7 &#x307E;&#x305F;&#x306F;&#x30B9;&#x30AD;&#x30FC; 1 &#x306B;&#x3088;&#x3063;&#x3066;&#x3067;&#x306F;&#x306A;&#x304F;&#x3002;&#x86CD;&#x5149;&#x767A;&#x751F;&#x30E2;&#x30C7;&#x30EB; &#x30DA;&#x30D7;&#x30C1;&#x30C9;&#x3068;&#x7D44;&#x63DB;&#x3048;&#x304A;&#x3092;&#x4F7F;&#x7528;&#x3057;&#x3066;&#x3001;&#x79C1;&#x305F;&#x3061;&#x30A2;&#x30B8;&#x30A2;&#x3072;&#x305A;&#x307F; E3E2 &#x30E2;&#x30C7;&#x30EB; &#x30DA;&#x30D7;&#x30C1;&#x30C9;&#x306F;&#x3001;&#x6700;&#x3082;&#x52B9;&#x7387;&#x7684;&#x306B;&#x98A8;&#x9234;&#x3068; PC5A &#x306B;&#x3088;&#x3063;&#x3066;&#x88C2;&#x304B;&#x3067;&#x3059;&#x89B3;&#x5BDF;&#x3002;&#x3053;&#x306E;&#x80F8;&#x306E;&#x8C37;&#x9593;&#x3082; CHIKV &#x611F;&#x67D3;&#x7D30;&#x80DE;&#x3067;&#x89B3;&#x5BDF;&#x3055;&#x308C;&#x3001;&#x98A8;&#x9234;&#x963B;&#x5BB3;&#x5264; decanoyl-RVKR-&#x30AF;&#x30ED;&#x30ED;&#x30E1;&#x30C1;&#x30EB; &#x30B1;&#x30C8;&#x30F3;&#x3067;&#x30D6;&#x30ED;&#x30C3;&#x30AF;&#x53EF;&#x80FD;&#x6027;&#x304C;&#x3042;&#x308A;&#x307E;&#x3059;&#x3002;&#x3053;&#x306E;&#x963B;&#x5BB3;&#x85AC;&#x30AF;&#x30ED;&#x30ED;&#x30AD;&#x30F3; CHIKV &#x524D;&#x8FF0;&#x306E;&#x3053;&#x306E;&#x30A6;&#x30A4;&#x30EB;&#x30B9;&#x306E;&#x81EA;&#x7136;&#x306A;&#x30BF;&#x30FC;&#x30B2;&#x30C3;&#x30C8;&#x3068;&#x3057;&#x3066;&#x7D30;&#x80DE;&#x578B;&#x7B4B;&#x82BD;&#x7D30;&#x80DE;&#x306E;&#x7D30;&#x80DE;&#x57F9;&#x990A;&#x3067;&#x306E;&#x62E1;&#x6563;&#x3092;&#x963B;&#x5BB3;&#x3059;&#x308B;&#x80FD;&#x529B;&#x3068;&#x6BD4;&#x8F03;&#x3057;&#x307E;&#x3057;&#x305F;&#x3002;&#x6211;&#x3005; &#x306E;&#x7D50;&#x679C;&#x306F;&#x51E6;&#x7406; CHIKV &#x7C92;&#x5B50;&#x306E;&#x98A8;&#x9234;&#x306E;&#x3088;&#x3046;&#x306A;&#x30D7;&#x30ED;&#x30C6;&#x30A2;&#x30FC;&#x30BC;&#x306E;&#x5F79;&#x5272;&#x3092;&#x793A;&#x3059;&#x305F;&#x3081;&#x3001;&#x3053;&#x306E;&#x611F;&#x67D3;&#x75C7;&#x3092;&#x6291;&#x5236;&#x3059;&#x308B;&#x65B0;&#x3057;&#x3044;&#x30A2;&#x30D7;&#x30ED;&#x30FC;&#x30C1;&#x3092;&#x30DD;&#x30A4;&#x30F3;&#x30C8;&#x3057;&#x307E;&#x3059;&#x3002;

&#x3072;&#x3068;&#x7B4B;&#x57F9;&#x990A;&#x306B;&#x304A;&#x3051;&#x308B;&#x30C1;&#x30AF;&#x30F3;&#x30B0;&#x30CB;&#x30A2; &#x30A6;&#x30A4;&#x30EB;&#x30B9;&#x611F;&#x67D3;&#x6291;&#x5236;&#x7D30;&#x80DE;&#x98A8;&#x9234;&#x963B;&#x5BB3;&#x5264;&#x306B;&#x3088;&#x308B;: E2 &#x8868;&#x9762;&#x7CD6;&#x86CB;&#x767D;&#x306E;&#x6210;&#x719F;&#x969C;&#x5BB3;&#x3002;

Chikungunya &#xBC14;&#xC774;&#xB7EC;&#xC2A4; (CHIKV)&#xC740; &#xC778;&#xAC04;&#xC758; &#xD2B9;&#xC9D5; polyarthralgia, &#xBC1C;&#xC5F4;, &#xADFC;&#xC721; &#xD1B5;&#xACFC; &#xB450;&#xD1B5;&#xC73C;&#xB85C; &#xAE09;&#xC131; &#xAC10;&#xC5FC; &#xC6D0;&#xC778; &#xBAA8;&#xAE30; &#xC804;&#xC1A1; Alphavirus. 2005 &#xB144; &#xC774;&#xD6C4;&#xC774; &#xBC14;&#xC774;&#xB7EC;&#xC2A4;&#xB294; &#xC804;&#xB840; &#xC5C6;&#xB294; &#xADDC;&#xBAA8;&#xC758; &#xC804;&#xC5FC;&#xBCD1; &#xD655;&#xC0B0;&#xC5D0; &#xB300; &#xD55C; &#xCC45;&#xC784; &#xB418;&#xC5C8;&#xC2B5;&#xB2C8;&#xB2E4;. &#xB2E4;&#xB978; alphaviruses&#xC640; &#xBE44;&#xC720;&#xC5D0; &#xC758;&#xD574; &#xC138;&#xD3EC; &#xD504;&#xB85C; &#xD14C;&#xC544; &#xBC14;&#xC774;&#xB7EC;&#xC2A4; &#xC131; &#xC804;&#xC870; &#xB2E8;&#xBC31;&#xC9C8; e1&#xC5D0;&#xB294; heterodimer&#xB85C; &#xC5F0;&#xACB0; &#xD558;&#xB294; &#xC218;&#xC6A9; &#xCCB4; &#xC758;&#xBB34;&#xC801;&#xC778; E2 &#xB2E8;&#xBC31;&#xC9C8;&#xC744; &#xC0DD;&#xC0B0; &#xD558;&#xB294; e3e2&#xC744; &#xCC98;&#xB9AC;&#xD560; &#xC218; &#xC788;&#xB294; &#xC0DD;&#xAC01;. &#xB0AE;&#xC740; Ph&#xC5D0; &#xB178;&#xCD9C;&#xC5D0; &#xC758;&#xD574; heterodimer&#xC758; &#xBD88;&#xC548; &#xBC14;&#xC774;&#xB7EC;&#xC2A4; &#xC131; &#xC735;&#xD569; &#xBC0F; &#xAC10;&#xC5FC; &#xC218; &#xC788;&#xC2B5;&#xB2C8;&#xB2E4;. &#xC6B0;&#xB9AC;&#xB294; proprotein convertases&#xC758; &#xB300;&#xD615; &#xD328;&#xB110; &#xC911; &#xB9C9; furin&#xC774; &#xC544;&#xB2C8;&#xB77C; PC5B &#xCC98;&#xB9AC;&#xD560; &#xC218; e3e2&#xB294; HRQRR(64)&#xC5D0;&#xC11C; &#xC544;&#xD504;&#xB9AC;&#xCE74; CHIKV &#xAE34;&#xC7A5;&#xC5D0;&#xC11C; &#xBCF4;&#xC5EC; / &#xC138;&#xC778;&#xD2B8; &#xC0AC;&#xC774;&#xD2B8;, &#xBC18;&#xBA74; &#xC544;&#xC2DC;&#xC544; &#xCD9C;&#xC2E0;&#xC758; CHIKV &#xBCC0;&#xD615; RRQRR(64)&#xC5D0; &#xBD99;&#xC5B4; / SI PC7 &#xB610;&#xB294; &#xC2A4;&#xD0A4; 1 &#xC544;&#xB2C8;&#xB77C; &#xC561;&#xCCB4;&#xB85C; &#xC6A9; &#xD574; furin, PC5A, PC5B, &#xBC0F; pace4&#xC5D0; &#xC758;&#xD574;. Fluorogenic &#xBAA8;&#xB378; &#xD3A9; &#xD2F0; &#xB4DC;&#xC640; &#xC7AC;&#xC870;&#xD569; convertases&#xB97C; &#xC0AC;&#xC6A9; &#xD558; &#xC5EC;, &#xC544;&#xC2DC;&#xC544; &#xC2A4;&#xD2B8;&#xB808;&#xC778; E3E2 &#xBAA8;&#xB378; &#xD3A9; &#xD2F0; &#xB4DC; furin&#xC640; pc5a&#xC5D0; &#xC758;&#xD574; &#xAC00;&#xC7A5; &#xD6A8;&#xC728;&#xC801;&#xC73C;&#xB85C; &#xD2B9;&#xD788;&#xB294; &#xAD00;&#xCC30; &#xD569;&#xB2C8;&#xB2E4;. &#xC774; &#xBD84;&#xC5F4; &#xB610;&#xD55C; CHIKV &#xAC10;&#xC5FC; &#xB41C; &#xC138;&#xD3EC;&#xC5D0;&#xC11C; &#xAD00;&#xCC30; &#xB418;&#xC5C8;&#xB2E4; &#xBC0F; furin &#xC5B5;&#xC81C;&#xC81C; decanoyl-RVKR-chloromethyl &#xCF00; &#xD1A4;&#xC5D0; &#xC758;&#xD574; &#xCC28;&#xB2E8; &#xB420; &#xC218; &#xC788;&#xC2B5;&#xB2C8;&#xB2E4;. &#xC774; &#xC5B5;&#xC81C;&#xC81C; &#xD074;&#xB85C;&#xB85C;&#xD038; CHIKV myoblast &#xC140; &#xBB38;&#xD654;&#xC5D0;&#xC11C; &#xD655;&#xC0B0;&#xC774; &#xBC14;&#xC774;&#xB7EC;&#xC2A4;&#xC758; &#xC790;&#xC5F0; &#xB300;&#xC0C1;&#xC73C;&#xB85C; &#xC124;&#xBA85; &#xC774;&#xC804;&#xC5D0; &#xC140; &#xD0C0;&#xC785;&#xC744; &#xC5B5;&#xC81C; &#xD558;&#xB294; &#xAE30;&#xB2A5;&#xC5D0; &#xB300; &#xD55C; &#xBE44;&#xAD50; &#xD588;&#xB2E4;. &#xC6B0;&#xB9AC;&#xC758; &#xACB0;&#xACFC; CHIKV &#xC785;&#xC790; &#xCC98;&#xB9AC;&#xC5D0; furin&#xC640; &#xAC19;&#xC740; &#xD504;&#xB85C; &#xD14C;&#xC544;&#xC758; &#xC5ED;&#xD560;&#xC744; &#xC124;&#xBA85; &#xD558; &#xACE0;&#xC774; &#xAC10;&#xC5FC;&#xC744; &#xC5B5;&#xC81C; &#xD558;&#xAE30; &#xC704;&#xD574; &#xC0C8;&#xB85C;&#xC6B4; &#xC811;&#xADFC;&#xC744; &#xC9C0;&#xC801;.

Furin &#xC5B5;&#xC81C;&#xC81C;&#xB85C; &#xC138;&#xD3EC;&#xAC00; &#xAD50;&#xC591; &#xC778;&#xAC04;&#xC758; &#xADFC;&#xC721;&#xC5D0;&#xC11C; Chikungunya &#xBC14;&#xC774;&#xB7EC;&#xC2A4; &#xAC10;&#xC5FC;&#xC758; &#xC5B5;&#xC81C;: E2 &#xD45C;&#xBA74; &#xB2F9;&#xB2E8;&#xBC31;&#xC9C8;&#xC758; &#xC131;&#xC219;&#xC758; &#xC7A5;&#xC560;.

Virus de chikungunya (CHIKV) es un Alphavirus transmitida por mosquitos que causa en los humanos una infecci&oacute;n aguda que se caracteriza por dolor de cabeza, fiebre, mialgia y polyarthralgia. Desde 2005 este virus ha sido responsable de un brote epid&eacute;mico de magnitud sin precedentes. Por analog&iacute;a con otros alphaviruses, se piensa que las proteasas celulares son capaces de procesar la prote&iacute;na viral precursor E3E2 para producir la prote&iacute;na de uni&oacute;n a receptores E2 que asocia como un heterod&iacute;mero con E1. Desestabilizaci&oacute;n de la heterod&iacute;mero por exposici&oacute;n a pH bajo permite la infecci&oacute;n y fusi&oacute;n viral. Mostramos que entre un amplio panel de convertasas del proprote&iacute;na furin membranosa sino tambi&eacute;n PC5B puede procesar E3E2 de cepas africanas CHIKV en el HRQRR(64) / ST sitio, mientras que una cepa CHIKV de origen asi&aacute;tico es dividida en RRQRR(64) / SI por furin membranosa y soluble, PC5A, PC5B y PACE4 pero no por PC7 o esqu&iacute;-1. Usando fluorogenic modelo p&eacute;ptidos y convertasas del recombinante, observamos que el p&eacute;ptido de cepa asi&aacute;tica E3E2 modelo es dividido m&aacute;s eficientemente por furin y PC5A. Este tambi&eacute;n fue observado en c&eacute;lulas infectadas por CHIKV y podr&iacute;a ser bloqueada por cetona de decanoyl-RVKR-clorometil furin inhibidor. Este inhibidor se compar&oacute; con cloroquina por su capacidad para inhibir la CHIKV separarse en cultivos celulares de mioblastos, un tipo de c&eacute;lulas, descrito como un objetivo natural de este virus. Nuestros resultados demuestran el papel de las proteasas furin-como en el procesamiento de part&iacute;culas CHIKV y se&ntilde;alan nuevos enfoques para inhibir esta infecci&oacute;n.

Inhibici&oacute;n de la infecci&oacute;n del virus de Chikungunya en m&uacute;sculo humano cultivado c&eacute;lulas por furin inhibidores: deterioro de la maduraci&oacute;n de la glicoprote&iacute;na de superficie de E2.

Chikungunya virus (CHIKV) is a mosquito-transmitted alphavirus that has been responsible for an epidemic outbreak of unprecedented magnitude in recent years. Since then, significant efforts have been made to better understand the biology of this virus, but we still have poor knowledge of CHIKV interactions with host cell components at the molecular level. Here we describe the extensive use of high-throughput yeast two-hybrid (HT-Y2H) assays to characterize interactions between CHIKV and human proteins. A total of 22 high-confidence interactions, which essentially involved the viral nonstructural protein nsP2, were identified and further validated in protein complementation assay (PCA). These results were integrated to a larger network obtained by extensive mining of the literature for reports on alphavirus-host interactions. To investigate the role of cellular proteins interacting with nsP2, gene silencing experiments were performed in cells infected by a recombinant CHIKV expressing Renilla luciferase as a reporter. Collected data showed that heterogeneous nuclear ribonucleoprotein K (hnRNP-K) and ubiquilin 4 (UBQLN4) participate in CHIKV replication in vitro. In addition, we showed that CHIKV nsP2 induces a cellular shutoff, as previously reported for other Old World alphaviruses, and determined that among binding partners identified by yeast two-hybrid methods, the tetratricopeptide repeat protein 7B (TTC7B) plays a significant role in this activity. Altogether, this report provides the first interaction map between CHIKV and human proteins and describes new host cell proteins involved in the replication cycle of this virus.

Mapping of Chikungunya virus interactions with host proteins identified nsP2 as a highly connected viral component.

Le virus Chikungunya (CHIKV) est un alphavirus transmises par la moustique qui a &eacute;t&eacute; responsable d&#39;une &eacute;pid&eacute;mie d&#39;une ampleur sans pr&eacute;c&eacute;dent ces derni&egrave;res ann&eacute;es. Depuis lors, des efforts importants ont &eacute;t&eacute; faits afin de mieux comprendre la biologie de ce virus, mais nous avons encore le manque de connaissances des interactions CHIKV avec des composants de cellule h&ocirc;te au niveau mol&eacute;culaire. Nous d&eacute;crivons ici la large utilisation du haut d&eacute;bit levure-hybride (HT-Y2H) essais pour caract&eacute;riser les interactions entre CHIKV et des prot&eacute;ines humaines. Un total de 22 haute-confiance des interactions, consistant essentiellement les prot&eacute;ines non structurales virale nsP2, ont &eacute;t&eacute; identifi&eacute;es et valid&eacute;s plus loin dans l&#39;analyse de compl&eacute;mentation prot&eacute;ique (PCA). Ces r&eacute;sultats ont &eacute;t&eacute; int&eacute;gr&eacute;s &agrave; un r&eacute;seau plus important obtenu par une vaste exploitation mini&egrave;re de la litt&eacute;rature pour les rapports sur les interactions h&ocirc;te-alphavirus. Afin d&#39;&eacute;tudier le r&ocirc;le des prot&eacute;ines cellulaires interagissant avec la nsP2, des exp&eacute;riences de g&egrave;ne silencieux ont &eacute;t&eacute; effectu&eacute;es dans les cellules infect&eacute;es par un CHIKV recombinant exprimant Renilla lucif&eacute;rase comme journaliste. A recueilli des donn&eacute;es a montr&eacute; que la ribonucl&eacute;oprot&eacute;ine nucl&eacute;aire h&eacute;t&eacute;rog&egrave;ne K (hnRNP-K) et ubiquilin 4 (UBQLN4) participent &agrave; la r&eacute;plication CHIKV in vitro. En outre, nous avons montr&eacute; que CHIKV nsP2 induit un cellulaire d&#39;arr&ecirc;t, comme d&eacute;j&agrave; signal&eacute; d&#39;autres alphavirus de l&#39;ancien monde et constate que parmi les partenaires de liaison identifi&eacute;s par des m&eacute;thodes de deux-hybride de levure, le tetratricopeptide r&eacute;p&egrave;te prot&eacute;ine 7 b (TTC7B) joue un r&ocirc;le important dans cette activit&eacute;. Au total, ce rapport fournit la premi&egrave;re carte de l&#39;interaction entre les CHIKV et les prot&eacute;ines humaines et d&eacute;crit les nouvelles prot&eacute;ines de cellules h&ocirc;tes impliqu&eacute;s dans le cycle de r&eacute;plication du virus.

Cartographie des interactions virus Chikungunya avec des prot&eacute;ines de l&#39;h&ocirc;te identifi&eacute; nsP2 comme une composante virale hautement connexe.

Chikungunya virus (CHIKV) &egrave; un alphavirus trasmesso da zanzare che &egrave; stato responsabile di un focolaio epidemico di ampiezza senza precedenti negli ultimi anni. Da allora, sono stati compiuti sforzi significativi per comprendere meglio la biologia di questo virus, ma abbiamo ancora scarsa conoscenza di interazioni CHIKV con componenti delle cellule ospite a livello molecolare. Qui descriviamo l&#39;uso estensivo di analisi di high-throughput lievito due-ibrido (HT-Y2H) per caratterizzare le interazioni tra CHIKV e proteine umane. Un totale di 22 interazioni di alta fiducia, che ha coinvolto essenzialmente il nsP2 virale proteine non strutturali, sono stati identificati e ulteriormente convalidato nell&#39;analisi della proteina di complementazione (PCA). Questi risultati sono stati integrati in una rete pi&ugrave; grande ottenuta dalla vasta mineraria della letteratura per i report sulle interazioni alphavirus-host. Per indagare il ruolo delle proteine cellulari interagendo con nsP2, sono stati eseguiti esperimenti del silenziamento del gene in cellule infettate da un CHIKV ricombinanti che esprimono Renilla luciferase come reporter. Dati raccolti hanno mostrato che ribonucleoproteina nucleare eterogenea K (hnRNP-K) e ubiquilin 4 (UBQLN4) partecipare alla replica di CHIKV in vitro. Inoltre, abbiamo dimostrato che CHIKV nsP2 induce un arresto cellulare, come precedentemente segnalato per altri alphaviruses del vecchio mondo e stabilito che tra i partner di associazione identificato da metodi di due-ibrido del lievito, del tetratricopeptide ripetere proteina 7B (TTC7B) svolge un ruolo significativo in questa attivit&agrave;. Complessivamente, questa relazione fornisce la prima mappa di interazione tra CHIKV e proteine umane e descrive il nuovo host cell proteine coinvolte nel ciclo di replicazione del virus.

Mappatura delle interazioni virus Chikungunya con proteine host identificato nsP2 come un componente virale altamente connessa.

Chikungunya virus (CHIKV) is a mosquito-transmitted pathogen responsible for an acute infection of abrupt onset, characterized by high fever, polyarthralgia, myalgia, headaches, chills, and rash. In 2006, CHIKV was responsible for an epidemic outbreak of unprecedented magnitude in the Indian Ocean, stressing the need for therapeutic approaches. Since then, we have acquired a better understanding of CHIKV biology, but we are still missing active molecules against this reemerging pathogen. We recently reported that the nonstructural nsP2 protein of CHIKV induces a transcriptional shutoff that allows the virus to block cellular antiviral response. This was demonstrated using various luciferase-based reporter gene assays, including a trans-reporter system where Gal4 DNA binding domain is fused to Fos transcription factor. Here, we turned this assay into a high-throughput screening system to identify small molecules targeting nsP2-mediated shutoff. Among 3040 molecules tested, we identified one natural compound that partially blocks nsP2 activity and inhibits CHIKV replication in vitro. This proof of concept suggests that similar functional assays could be developed to target other viral proteins mediating a cellular shutoff and identify innovative therapeutic molecules.

A phenotypic assay to identify Chikungunya virus inhibitors targeting the nonstructural protein nsP2.

The capacity of a virus to cross species barriers is determined by the development of bona fide interactions with cellular components of new hosts, and in particular its ability to block IFN-Î±/Î² antiviral signaling. Tioman virus (TioV), a close relative of mumps virus (MuV), has been isolated in giant fruit bats in Southeast Asia. Nipah and Hendra viruses, which are present in the same bat colonies, are highly pathogenic in human. Despite serological evidences of close contacts between TioV and human populations, whether TioV is associated to some human pathology remains undetermined. Here we show that in contrast to the V protein of MuV, the V protein of TioV (TioV-V) hardly interacts with human STAT2, does not degrade STAT1, and cannot block IFN-Î±/Î² signaling in human cells. In contrast, TioV-V properly binds to human STAT3 and MDA5, and thus interferes with IL-6 signaling and IFN-Î² promoter induction in human cells. Because STAT2 binding was previously identified as a host restriction factor for some Paramyxoviridae, we established STAT2 sequence from giant fruit bats, and binding to TioV-V was tested. Surprisingly, TioV-V interaction with STAT2 from giant fruit bats is also extremely weak and barely detectable. Altogether, our observations question the capacity of TioV to appropriately control IFN-Î±/Î² signaling in both human and giant fruit bats that are considered as its natural host.

The V protein of Tioman virus is incapable of blocking type I interferon signaling in human cells.

La capacit&eacute; d&#39;un virus de franchir des barri&egrave;res d&#39;esp&egrave;ces est d&eacute;termin&eacute;e par le d&eacute;veloppement de v&eacute;ritables interactions avec des composants cellulaires de nouveaux h&ocirc;tes et en particulier sa capacit&eacute; &agrave; bloquer l&#39;IFN-&alpha;/&beta; de signalisation antivirales. Tioman virus (TioV), un proche parent du virus ourlien (MuV), a &eacute;t&eacute; isol&eacute; chez des roussettes g&eacute;antes en Asie du sud-est. Virus Nipah et Hendra, qui sont pr&eacute;sents dans les m&ecirc;mes colonies de chauve-souris, sont hautement pathog&egrave;nes chez l&#39;homme. Malgr&eacute; les &eacute;vidences s&eacute;rologiques des contacts &eacute;troits entre TioV et les populations humaines, si TioV est associ&eacute; &agrave; une pathologie humaine reste ind&eacute;termin&eacute;e. Nous montrons ici que contrairement &agrave; la prot&eacute;ine V de MuV, la prot&eacute;ine V de TioV (TioV-V) peine interagit avec STAT2 humain, ne d&eacute;grade pas STAT1 et ne peut pas bloquer l&#39;IFN-&alpha;/&beta; de signalisation dans les cellules humaines. En revanche, TioV-V lie correctement &agrave; humain STAT3 et MDA5 et interf&egrave;re donc avec signalisation d&#39;IL-6 et induction promoteur IFN-&beta; dans les cellules humaines. Parce que STAT2 liaison a d&eacute;j&agrave; &eacute;t&eacute; identifi&eacute;e comme un facteur de restriction d&#39;h&ocirc;te pour des Paramyxoviridae, nous avons &eacute;tabli la s&eacute;quence STAT2 de roussettes g&eacute;antes, et liaison &agrave; TioV-V a &eacute;t&eacute; mis &agrave; l&#39;essai. &Eacute;tonnamment, TioV-V interaction avec STAT2 de roussettes g&eacute;antes est &eacute;galement extr&ecirc;mement faible et &agrave; peine d&eacute;celable. Au total, nos observations en question la capacit&eacute; de TioV de contr&ocirc;ler ad&eacute;quatement IFN-&alpha;/&beta; la signalisation dans les roussettes g&eacute;antes tant humaines qui sont consid&eacute;r&eacute;s comme son h&ocirc;te naturel.

La prot&eacute;ine V de Tioman virus est incapable de bloquer les signaux dans les cellules humaines de l&#39;interf&eacute;ron de type I.

La capacit&agrave; di un virus di attraversare le barriere di specie &egrave; determinata dallo sviluppo di bona fide interazioni con componenti cellulari dei nuovi padroni di casa e in particolare la capacit&agrave; di bloccare la segnalazione antivirale IFN-&alpha;/&beta;. Tioman virus (TioV), un parente stretto del virus della parotite (MuV), &egrave; stata isolata in gigante pipistrelli della frutta nel sud-est asiatico. Virus Nipah e Hendra, che sono presenti nella stessa colonie di pipistrelli, sono altamente patogeni umani. Nonostante le prove sierologiche di stretti contatti tra popolazioni umane e TioV, se TioV &egrave; associato a qualche patologia umana rimane indeterminato. Qui vi mostriamo in contrasto con la proteina V di MuV, la proteina V di TioV (TioV-V) difficilmente interagisce con STAT2 umana, non degradano STAT1 che non pu&ograve; bloccare l&#39;IFN-&alpha;/&beta; segnalazione nelle cellule umane. In contrasto, TioV-V correttamente associa all&#39;umano STAT3 e MDA5 e quindi interferisce con la segnalazione di IL-6 e induzione promotore IFN-&beta; nelle cellule umane. Perch&eacute; STAT2 associazione era identificato come un fattore di limitazione di host per alcuni Paramyxoviridae, abbiamo stabilito la sequenza STAT2 gigante pipistrelli della frutta, e associazione TioV-v &egrave; stato testato. Sorprendentemente, TioV-V interazione con STAT2 da gigante pipistrelli della frutta &egrave; anche estremamente debole e appena rilevabile. Complessivamente, le nostre osservazioni domanda la capacit&agrave; della TioV per controllare adeguatamente IFN-&alpha;/&beta; segnalazione nei pipistrelli della frutta sia umani e giganti che sono considerate come suo ospite naturale.

La proteina V di Tioman virus &egrave; incapace di blocco tipo interferone segnalazione nelle cellule umane.

Chikungunya virus (CHIKV) &egrave; un arbovirus recentemente riemersa che innesca autophagy. Qui, ci mostra che CHIKV interagisce con i componenti della macchina autophagy durante il suo ciclo di replicazione, inducendo un effetto citoprotettivo. Il autophagy recettore p62 protegge le cellule dalla morte associando ubiquitarie capside e targeting per esso a autophagolysosomes. Di contrasto, il recettore umano autophagy NDP52-ma non il mouse orthologue-interagisce con le proteine non-strutturali nsP2, promuovendo cos&igrave; la replicazione virale. Questi risultati evidenziano i ruoli distinti di p62 e NDP52 nell&#39;infezione virale e identificano NDP52 come un fattore cellulare che conti per specificit&agrave; di specie CHIKV.

Specie-impatto delle macchine sull&#39;infezione da virus Chikungunya autophagy.

Chikungunya virus (CHIKV) is a recently re-emerged arbovirus that triggers autophagy. Here, we show that CHIKV interacts with components of the autophagy machinery during its replication cycle, inducing a cytoprotective effect. The autophagy receptor p62 protects cells from death by binding ubiquitinated capsid and targeting it to autophagolysosomes. By contrast, the human autophagy receptor NDP52-but not its mouse orthologue-interacts with the non-structural protein nsP2, thereby promoting viral replication. These results highlight the distinct roles of p62 and NDP52 in viral infection, and identify NDP52 as a cellular factor that accounts for CHIKV species specificity.

Species-specific impact of the autophagy machinery on Chikungunya virus infection.

(CHIKV) le virus Chikungunya est un arbovirus r&eacute;cemment r&eacute;apparu qui d&eacute;clenche l&#39;autophagie. Ici, nous montrons que CHIKV interagit avec les composants de la machine de l&#39;autophagie pendant son cycle de r&eacute;plication, induisant un effet cytoprotecteur. L&#39;autophagie r&eacute;cepteur p62 prot&egrave;ge les cellules de la mort en se liant ubiquitin&eacute;e capside et ciblant &agrave; autophagolysosomes. En revanche, le r&eacute;cepteur humain autophagie NDP52-mais pas ses orthologue de la souris-interagit avec la prot&eacute;ine non structurale nsP2, ce qui favorise la r&eacute;plication virale. Ces r&eacute;sultats soulignent les r&ocirc;les distincts de p62 et NDP52 dans l&#39;infection virale et d&#39;identifient NDP52 comme un facteur cellulaire qui repr&eacute;sente sp&eacute;cificit&eacute; d&#39;esp&egrave;ce CHIKV.

Effets sp&eacute;cifiques &agrave; l&#39;esp&egrave;ce de la machinerie de l&#39;autophagie sur l&#39;infection par le virus Chikungunya.

Searching for stimulators of the innate antiviral response is an appealing approach to develop novel therapeutics against viral infections. Here, we established a cell-based reporter assay to identify compounds stimulating expression of interferon-inducible antiviral genes. DD264 was selected out of 41,353 compounds for both its immuno-stimulatory and antiviral properties. While searching for its mode of action, we identified DD264 as an inhibitor of pyrimidine biosynthesis pathway. This metabolic pathway was recently identified as a prime target of broad-spectrum antiviral molecules, but our data unraveled a yet unsuspected link with innate immunity. Indeed, we showed that DD264 or brequinar, a well-known inhibitor of pyrimidine biosynthesis pathway, both enhanced the expression of antiviral genes in human cells. Furthermore, antiviral activity of DD264 or brequinar was found strictly dependent on cellular gene transcription, nuclear export machinery, and required IRF1 transcription factor. In conclusion, the antiviral property of pyrimidine biosynthesis inhibitors is not a direct consequence of pyrimidine deprivation on the virus machinery, but rather involves the induction of cellular immune response.

Marianne Lucas-Hourani

Unité de Génomique Virale et Vaccination,
Virology Department,
Unité de Génomique Virale et Vaccination, Virology Department

Institut Pasteur, CNRS UMR3569

High-throughput Screening for Broad-spectrum Chemical Inhibitors of RNA Viruses

Marianne Lucas-Hourani

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}Unité de Génomique Virale et Vaccination,Virology Department,Unité de Génomique Virale et Vaccination, Virology Department

Institut Pasteur, CNRS UMR3569

High-throughput Screening for Broad-spectrum Chemical Inhibitors of RNA Viruses

Unité de Génomique Virale et Vaccination,
Virology Department,
Unité de Génomique Virale et Vaccination, Virology Department