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Investigating the structural role of the Hepatitis B virus core protein C-terminal domain in assembly and maturation using solid-state NMR

Objective

We will seek in this project to reveal structural and dynamic features of the C-terminal domain (CTD) of the Hepatitis B virus (HBV) core protein which encapsidates the viral genome. Each year, over 750,000 deaths are due to serious liver diseases caused by chronic HBV infection. Hepatitis B is, despite the availability of a vaccine, still a major health problem. The HBV virus particle is composed of an outer lipid-protein envelope and a core protein of 20 kDa that self-assembles into icosahedral nucleocapsids. The nucleocapsid encloses the pregenomic RNA (pgRNA) that is then reverse-transcribed into DNA by the viral polymerase (Pol). During pgRNA encapsidation, the CTD is thought to interact with the RNA. Despite the essential role played by the CTD in the regulation of the viral life cycle through various conformational states and phosphorylation patterns, it escapes today to classical structural biology approaches due to its dynamic nature. We will use solid-state NMR to bring new insights into the precise conformation and localization of the CTD along various steps of the viral life cycle. Sample preparation protocols of the core protein have recently been established by the applicant in the host laboratory using bacterial expression systems. The first NMR spectra of the nucleocapsid look highly promising. Further experiments will be recorded to assign the resonances and identify the CTD signals. Different phosphorylation states and binding partners, as the pgRNA, will be explored. Finally, paramagnetic tags will be used to obtain information about the positioning of the CTD.
Our work shall contribute to expand the present molecular description of the HBV life cycle to include the role of the CTD. Our results will help to understand, and ultimately to target, its central role in HBV infection.

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Topic(s)

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Funding Scheme

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MSCA-IF-EF-ST - Standard EF

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Call for proposal

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(opens in new window) H2020-MSCA-IF-2016

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Coordinator

CENTRE NATIONAL DE LA RECHERCHE SCIENTIFIQUE CNRS
Net EU contribution

Net EU financial contribution. The sum of money that the participant receives, deducted by the EU contribution to its linked third party. It considers the distribution of the EU financial contribution between direct beneficiaries of the project and other types of participants, like third-party participants.

€ 143 603,35
Total cost

The total costs incurred by this organisation to participate in the project, including direct and indirect costs. This amount is a subset of the overall project budget.

€ 185 076,00
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