Molecular analysis of antiviral RNA granules induced by vaccinia virus infection

This sandwich placement will be based in the lab ofMichael Way.

Project background and description

Poxviruses are a large family of enveloped DNA viruses that can cause serious diseases in humans. Notable members of the family include the etiological agents of mpox (formerly monkeypox) and smallpox. Vaccinia virus,the prototype poxvirus, is widely used to investigate the complex replication cycle of these viruses, which occurs exclusively in the cytoplasm of infected cells.

During vaccinia infection, RNA granules form around viral replication sites as part of the host mediated stress response.These infection-induced granules function to suppress viral gene expression by inhibiting mRNA translation. They contain viral double-stranded RNA (dsRNA) and canonical stress granule markers, such as G3BP1, which are typically associated withcytoplasmic RNA-protein aggregates that form in non-infected during a variety of different stresses. We recently demonstrated that the tRNA ligase complex is present in vaccinia-induced antiviral granules, however, their full molecular composition remains to be fully elucidated.

This project aims to determine the complete protein and RNA composition of antiviral granules formed during vaccinia virus infection. We will use proximity labelling and pulldown approaches with tagged G3BP1 andDDX1 stably expressed in cells, to biotinylate proteins and RNA molecules in viral-induced granules. Biotinylated components will be isolated via streptavidin-based purification and subsequently identified via mass spectrometry and RNA sequencing. Hits will be validated using imaging approaches on fixed and live vaccinia infected cells.

Candidate background

The post holder should embody and demonstrate the Crick ethos and ways of working: bold, open and collegial. The candidate must be registered at a UK Higher Education Institution, studying in the UK and must have completed a minimum of two years’ undergraduate study in a relevant discipline, and on track to receive a final degree grade of 2:1 or 1.In addition, they should be able demonstrate the following experience and key competencies:

• This project is suited for a student in biological sciences / biochemistry / cell biology with an interest in host-pathogen interactions and a desire to develop a broad skill set in molecular biology, RNA biology, cell biology and biochemical techniques
• Good knowledge in relevant scientific area(s)
• Good written and spoken communication skills
• Ability to work independently and also capable of interacting within a group

References

• Nonredundant roles of topoisomerase 2α and 2β in the cytosolic replica Nucleic Acids Research 53: gkaf566. PubMed abstract
• Activation of stress response pathways promotes formation of antiviral granules and restricts virus replication. Molecular and Cellular Biology 34: 2003-2016. PubMed abstract