Project #15049
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#15049 : Determination of RABV RNAs signatures recognized by RLRs
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Name of Applicant : Wahiba Aouadi
Date of application : 26-02-2020
Unit : Lyssavirus Dynamics and Host Adaptation
Location : Lwoff, 1st floor
Phone : 3927
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Project context and summary :

Rabies virus (RABV) is a causative agent of lethal neurological disease. It presents a public health threat in the world resulting in more than 59,000 human deaths every year around the world. RABV possesses negative strand RNA genome, 11.9 kb, encoding five viral proteins: Nucleoprotein (N), Phosphoprotein (P), Matrix protein (M), Glycoprotein (G) and polymerase or Large protein (L). Structural modelling of L protein suggested that L contained different conserved domains: i) RdRp for RNA transcription and replication, ii) capping domain and iii) methyltransferase domain (MTase) mediating the transfer of methyl molecule on mRNA cap structure. The detection of viral presence in a host cell is mediated by pattern recognition receptors (PRRs) that sense viral pathogen-associated molecular patterns (PAMPs). The cytosolic receptors include RIG-I-like receptors (RLRs). Three RLRs are known: RIG-I, MDA5 and LGP2. Upon ligand recognition, MDA5 and RIG-I activate signaling pathways leading to the production of pro-inflammatory cytokines, including type-I interferon (IFN), and the establishment of an antiviral state in the host. We have recently performed RLR siRNA knockdown experiments suggesting that upon RABV infection RIG-I is the major RLR implicated in RABV RNA sensing and IFN induction. However, the precise nucleic acid sequence of RABV RNA sensed by RLRs remains unknown. In order to determine RABV RNAs signatures recognized by RIG-I and MDA5, in collaboration with A. Komarova (IP), we have applied a novel ribonucleoproteomic approach that consists in affinity purification of tagged RLRs followed by extraction of associated RNA molecules. This approach has recently provided comparative views on viral RLR ligands in the context of infection with Measles, Chikungunya and Dengue viruses. We are now ready to perform NSG analysis of RIG-I- and MDA5- specific RNA ligands purified from RABV infected cells.

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Project Type : Medium
Status : Pending

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