A structural and functional understanding Influenza virus mRNA production in the context of the transcriptionally functional ribonucleoprotein (RNP) particle.

Descripción del proyecto

Un complejo ARN-proteína es fundamental para la transcripción del virus de la gripe

Los virus de la gripe son virus de ARN que representan una gran amenaza para la salud de las personas a través de epidemias anuales y pandemias puntuales. Comprender la biología de los virus de la gripe es fundamental para diseñar fármacos antivirales y vacunas. El proyecto RNP, financiado con fondos europeos, investigará los complejos de ribonucleoproteínas (RNP, por sus siglas en inglés) implicados en la transcripción del ARN genómico viral en ARNm. El objetivo de los investigadores es definir los cambios dinámicos en la estructura de las RNP como procedimientos de transcripción. Los resultados proporcionarán información bioquímica y molecular fundamental sobre los mecanismos de replicación de los virus de la gripe, lo cual abrirá nuevas vías de investigación de fármacos contra dicha enfermedad. Además, las metodologías desarrolladas se podrían aplicar para estudiar complejos de ribonucleoproteínas esenciales de otros virus de ARN patógenos.

Objetivo

Influenza is a major public health burden, with seasonal outbreaks contributing significantly to mortality worldwide, and the emergence of pandemic strains remaining an ever-present threat. Influenza drug and vaccine conception efforts are aided by a thorough understanding of its molecular biology.

A key aspect of the influenza lifecycle is the production of capped and poly-adenylated messenger RNA by the heterotrimeric influenza polymerase (FluPol). Ground-breaking work performed by the Cusack lab, has described with residue-resolution detail, the FluPol structures that form during transcription of short, non-nucleoprotein (NP) bound viral RNAs (vRNAs). However, influenza transcription in vivo occurs within the ribonucleoprotein (RNP) particle and does not utilise naked genome segments. The viral RNP (vRNP) is a super-helical complex composed FluPol bound at the conserved 3′ and 5′ ends of a vRNA, which is coated with NP. The current low-resolution structures provide little information about the molecular details of vRNP function, particularly, how NPs interact with FluPol and the vRNA template.

Via an inter-disciplinary approach, I will utilise cryo-electron microscopy methods, transcription assays and single-molecule fluorescence, to obtain the first high resolution structure of a dynamic influenza vRNP, with a particular focus on the spatial organisation of NPs relative to FluPol. In addition to this work facilitating future influenza drug research, it will provide a basis to investigate the vRNP during other lifecycle stages and act as proof-of-principle for study of other viral protein-RNA complexes, such as those from corona-, arena- and bunyaviruses.

Work will be performed in the groups of Stephen Cusack and Olivier Duss based at EMBL Grenoble and Heidelberg, respectively. Here, I will have access to world-leading facilities and training opportunities, supporting my growth as an independent researcher and an expert in RNA virus structural biology.