Fundamental biology of E2F transcription factors: investigating the role of arginine methylation

Growth and division of animal cells requires gene expression to be turned on and off in a coordinated fashion. The process of cell division is called the cell cycle, during which chromosomes are first copied and then one cell divides into two. A major point of control occurs during the first G1 phase, where cells commit to replicate their chromosomes and divide or alternatively exit the cell cycle. In normal cells, the cell cycle is a tightly regulated process, but frequently becomes awry in human disease. The key proteins involved, the retinoblastoma tumour suppressor protein pRb and its crucial target E2F, govern progress through G1, and the commitment of cells to divide. This research addresses the mechanisms which are responsible for normal cell cycle regulation focussing on new levels of enzymatic control which influence the regulatory properties of E2F, focussing on the E2F-1 subunit, which exhibits some profoundly interesting and unusual biological properties; for example, it is able to prompt cell growth and survival, and conversely stimulate cell death. Nothing is really known about the molecular mechanisms which control these contrasting biological outcomes of E2F-1 activity, which is the focus of this application. Specifically, the programme will dissect the molecular mechanisms and cellular processes through which E2F-1 exerts its biological effects in cells. We will concentrate on the mechanisms which endow E2F with its opposing biological properties, and probe the cellular pathways and molecular processes that are responsible. Our experimental approach uses advanced methodologies to address important questions on how E2F exerts its effects at the genome and proteome level, and we work with a group of expert collaborators to achieve these aims. Our research has the overarching aim of delivering a detailed understanding of the role of E2F in normal cells, and the mechanisms which are responsible for controlling its distinct biological roles. The study is very likely to illuminate new mechanisms and information that is relevant to human disease.

The E2F family of transcription factors is a conserved group of master regulators that function in cell cycle regulated gene expression. The E2F family is a critical target for the pRb tumour suppressor to exert negative growth control, and the pathway is subject to aberrant control in many human diseases. Our understanding of the E2F family, which is most advanced for the E2F-1 subunit, has shown that E2F subunits exert different biological effects reflecting a cell and tissue context dependency. Most notably, E2F-1 is endowed with paradoxical biological activities as it can facilitate cell cycle progression and, in some physiological circumstances, prompt the opposing outcome of apoptosis. Our proposal aims to systematically dissect the molecular processes and cellular pathways that control the biological properties of E2F activity, with a particular focus on the role of a newly described type of post-translational modification which impacts on the biological output of E2F-1. Thus, we now know that arginine (R) methylation (me) is a crucial level of control on E2F-1, where the type of methylation event acts as a molecular switch, to trigger either cell growth or cell death. Significantly, meR mediated by the PRMT5 enzyme prompts cell growth, and the meR mark is 'read' by chromatin-associated tudor domain (TD) proteins. Our key objective is to understand at the molecular mechanistic level how the two types of meR mark control E2F-1 activity, and relate this information to the biological properties of E2F-1 in normal cellular systems. Our technical approach is multi-disciplinary, combining advanced genomics and proteomics technologies with gene editing, cell biology and biochemistry, to deliver new fundamental knowledge relevant to our understanding of the cell cycle and control of cell fate.

Our target audience is broad, including the wider public, the public sector, charities and commercial/private sector. We will raise awareness of our work and its impact for the general non-technical public through open days and media engagement. We shall aim to influence decision makers and opinion formers of the positive impact of our work and alert government agencies and charities to the important role they play in research funding through workshops and focussed end-user group discussions. We shall continue to alert and educate the private sector to the commercial benefits and long-term economic importance of our research, through presentations at trade and investor meetings. The research has the potential to enhance the quality of life by illuminating fundamental biological processes about how normal cells grow and divide, and thereby in the longer term provide new pathways and mechanisms that can be translated into innovative drug discovery and therapeutic approaches eventually leading to healthier lives . At the economic level, our research will foster new intellectual property, provide the basis for novel spin-out companies and pharmaceutical collaborations, and drive the economic competitiveness of the UK. Societal impact will reflect an improved public awareness of the importance of fundamental biological research for a better understanding of disease processes. Staff working on the project will become highly skilled and thereby strengthen the UK's competitive position, and develop improved communication techniques by contributing to the impact strategy and thereby realise improved career development opportunities. In the longer term, our research will impact on health and quality of life, by providing the foundation for new therapeutic approaches.
Our studies will be published in high impact scientific journals, although where relevant we will issue press releases with a non-scientific lay explanation, to accentuate public engagement and uptake. On several recent occasions, we have made radio and TV appearances to maximise press coverage, using the Medical Sciences Press Office, and have frequently appeared in national newspapers, which we shall continue in the future programme. The Department has a number of Open Days each year where the public is invited to attend. The laboratory has and will continue to present our work at Open Days in a fashion that maximises accessibility to increase public understanding of our work, and its value for new medical discoveries and the nation's health. All our publications and research outputs are, wherever possible, open access. New materials and reagents are made freely available to the research community, and large data sets deposited in relevant data bases using centralised standards compliant with data for public access. Results that have intellectual property (IP) implications will be discussed in advance with ISIS (the intellectual property arm of Oxford University) before public disclosure. Of further relevance, we run the Oxford Branch of the British Science Association. The branch runs monthly SciBars, has its own website (www.oxfordscibar.com), and actively engages in the local and wider scientific community through not only running monthly events, but through podcasting their events (freely available via the website or iTunes), engaging on twitter and other social media platforms as well as through collaborations with Oxford University Press and Science Oxford. The events are run at the pub and are aimed at a general audience, and past highlights include Colin Blakemore, Peter Atkins, Jocelyn Bell Burnell and Alison Woollard (Royal Institution Christmas Lecturer).