Arenavirus RNPs and Specialized Ribosomes

Lead Research Organisation: University of Leeds
Department Name: Astbury Centre

Abstract

Background
The segmented negative stranded RNA viruses are a wide group of viruses that includes three families namely, bunyaviruses, arenaviruses and orthomyxoviruses. Together these families cause widespread disease in humans animals and plants that pose major threats from disease and economic loss. As with all these families, arenaviruses possesses an RNA genome that is wrapped up in a viral nucleocapsid protein (NP) to form ribonucleocapsids (RNPs), which are essential for virus gene expression and assembly, making RNP formation a potential drug target. In addition, arenaviruses selectively package ribosomes into virus particles raising the intriguing possibility that the virus uses specialised ribosomes to favour viral protein translation over that of the host, and to expedite the progression of virus growth.
This project will investigate the structure of the NP protein and how it recognises viral RNA, the structure and assembly of RNPs, the structure of virus particles and how they assemble in cells, and analyse the profile of ribosomes associated with viral transcription and translation.

Objectives
1) Purify recombinant NP protein from Lujo, Pichinde or LCM virus and solve the crystal structure with/out RNA.
2) Purify RNPs from live Arenavirus and solve the RNP structure by cryo-EM.
3) Profile ribosomes from live Arenavirus and assess whether these viruses are using specialised ribosomes.
4) Visualise whole virus particles and virus assembling in cells by cryo-EM tomography.

Novelty
Little is known about the variability in N protein structure across the Arenaviruses, and this may impact potential for design of pan-Arena therapeutics against N. The concept of specialised ribosomes for specific jobs is also an emerging theme.

Timeliness
The structure of negative sense RNA virus RNPs is a hot topic at the moment, and the novelty of ribosome profiling also adds timeliness

Experimental Approach
Cell culture, recombinant protein expression, X-ray crystallography, cryo-EM and tomography.

Publications

10 25 50

Studentship Projects

Project Reference Relationship Related To Start End Student Name
BB/M011151/1 30/09/2015 29/09/2023
1780151 Studentship BB/M011151/1 30/09/2016 30/03/2021 Amelia Shaw