Ligands of MDA5 and RIG-I in Measles Virus-Infected Cells


RIG-I-like receptors (RLRs) are helicase-like molecules that detect cytosolic RNAs that are absent in the non-infected host. Upon binding to specific RNA patterns, RLRs elicit a signaling cascade that leads to host defense via the production of antiviral molecules. To understand how RLRs sense RNA, it is important to characterize the nature and origin of RLR-associated RNA from virus-infected cells. While it is well established that RIG-I binds 5′-triphosphate containing double-stranded RNA, the in vivo occurring ligand for MDA5 is poorly characterized. A major challenge in examining MDA5 agonists is the apparently transient interaction between the protein and its ligand. To improve the stability of interaction, we have used an approach to crosslink MDA5 to RNA in measles virus-infected cells. The virus-infected cells were treated with the photoactivatable nucleoside analog 4-thiouridine, which is incorporated in newly synthesized RNA. Upon 365 nm UV light exposure of living cells, a covalent linkage between the labeled RNA and the receptor protein is induced, resulting in a higher RNA recovery from RLR immunoprecipitates. Based on next generation sequencing, bioinformatics and in vitro approaches, we observed a correlation between the AU-composition of viral RNA and its ability to induce an MDA5-dependent immune response.


Vyšlo v časopise: Ligands of MDA5 and RIG-I in Measles Virus-Infected Cells. PLoS Pathog 10(4): e32767. doi:10.1371/journal.ppat.1004081
Kategorie: Research Article
prolekare.web.journal.doi_sk: 10.1371/journal.ppat.1004081

Souhrn

RIG-I-like receptors (RLRs) are helicase-like molecules that detect cytosolic RNAs that are absent in the non-infected host. Upon binding to specific RNA patterns, RLRs elicit a signaling cascade that leads to host defense via the production of antiviral molecules. To understand how RLRs sense RNA, it is important to characterize the nature and origin of RLR-associated RNA from virus-infected cells. While it is well established that RIG-I binds 5′-triphosphate containing double-stranded RNA, the in vivo occurring ligand for MDA5 is poorly characterized. A major challenge in examining MDA5 agonists is the apparently transient interaction between the protein and its ligand. To improve the stability of interaction, we have used an approach to crosslink MDA5 to RNA in measles virus-infected cells. The virus-infected cells were treated with the photoactivatable nucleoside analog 4-thiouridine, which is incorporated in newly synthesized RNA. Upon 365 nm UV light exposure of living cells, a covalent linkage between the labeled RNA and the receptor protein is induced, resulting in a higher RNA recovery from RLR immunoprecipitates. Based on next generation sequencing, bioinformatics and in vitro approaches, we observed a correlation between the AU-composition of viral RNA and its ability to induce an MDA5-dependent immune response.


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Hygiena a epidemiológia Infekčné lekárstvo Laboratórium

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