BBSRC EASTBIO DTP - Dissecting the proviral functions of Jmjd6 in influenza A virus infection

Influenza A viruses (IAV) cause annual epidemics and are a major threat for public health and the global economy. To grow inside cells the virus relies on host cell machinery to promote viral gene expression and virus replication. Although many high-throughput screens have identified host proteins that are essential for virus replication there is still a limited understanding of how these factors contribute to a successful virus life cycle and how they are co-opted by the virus.

We have recently identified that the jumonji domain containing protein 6 (Jmjd6) is a cellular host factor required for viral growth. Jmjd6 is a nuclear hydroxylase which interacts with transcription initiation factors and spliceosomal components and thereby regulates host gene expression (Webby et al. 2009, Liu et al. 2013). Cells that lack Jmjd6 show defects in IAV gene and protein expression and show consequently reduced production of new virions. The objective of this PhD project is to investigate at which step of the viral life cycle Jmjd6 is required for viral growth. Potentially Jmjd6 could impact on viral gene expression by influencing transcriptional initiation, processing of viral mRNAs (pre-mRNA splicing) or by affecting shutteling of viral ribonucleoprotein complexes into the nucleus. This project will use CRISPR-Cas9 edited Jmjd6-deficient cells and Jmjd6 expression reporter constructs as tools to functionally dissect these different possibilities for Jmjd6-virus interactions. This will involve imaging of viral infections by confocal microscopy, immunoprecipitation of host and virus candidate interactor proteins, and assays that investigate viral and host RNA expression. For this we are offering training in biochemistry, virology, molecular biology and in genetics of host-pathogen interactions. We have recently generated conditional Jmjd6 knockout mice. This new allele will allow us to delete Jmjd6 in IAV host target cells and to characterise the importance of the protein for the infection process in vivo. We will use imaging technologies to track and compare the dynamics of IVA infection in Jmjd6-deficient and wild type mice. Training will be provided in two-photon laser scanning intravital microscopy, bioluminescence imaging, and histopathology.