Post-transcriptional regulation of influenza A virus RNA

This research proposal aims to significantly alter our understanding of the critical role post-transcriptional processes play in the influenza A virus (IAV) life cycle. Post-transcriptional regulation of cellular mRNAs has seen a lot of research interest in recent years, including projects looking at the effects of RNA modifications and ribosome specialisation. However, much less attention has been paid to the effects these processes have on the viral life cycle. This project focuses on the post-transcriptional regulation of both IAV mRNAs and negative strand vRNAs. However, outcomes of this work will have a profound effect on our perceptions of the regulatory processes affecting a wide range of viral RNAs. In fact, by better understanding the roles of these processes on viral RNAs, such as IAV, we can also uncover novel functions on cellular mRNAs. This project comprises 5 work packages with 11 intermediate goals. We will first identify the locations of various modifications present on IAV RNAs across multiple strains in both human and avian infected cells, significantly expanding on our current understanding, while exploring the potential for species-specific adaptions. Through mutagenesis and RNA capture techniques, we will evaluate how these modifications affect RNA characteristics and what effector proteins are involved in these processes. We will also use this information to determine the composition of ribosomes actively translating IAV mRNAs and evaluate whether specialised ribosomes are involved in the normal IAV life cycle. Finally, we will focus on the roles of RNA modifications on vRNAs, which should be quite distinct from mRNAs, and the host proteins that specifically bind, or are blocked from binding, sites of modification. This is an ambitious, multifaceted project that will have a direct impact on our understanding of IAV biology, and also provide novel insights of value to multiple disciplines including virology, RNA biology and protein translation.