The way in which bacteria divide in order to grow is a highly-coordinated process and requires a complex choreography of many proteins, working inside and outside the cell membrane. Many of these proteins have a functional relationship with the synthesis and coordination of the external cell shape determining polymer called peptidoglycan (PG). This polymer provides a structural scaffold for many cellular processes as well as mechanical strength and protection, which requires modification during the process of cell division. A critical point occurs during cell division when the "old" cell wall PG must be degraded to allow separation of newly synthesized cells and this function is provided by a variety of PG hydrolases associated with the cell division FtsE-FtsX protein complex. At Warwick we have recently contributed to a new understanding of how this process occurs in rod shaped, gram negative bacteria. However, the situation in respiratory infection associated, ovoid-shaped gram positive bacterial pathogens, including Streptococcus pneumoniae is unclear at present. Notably there is a direct interaction and functionally essential interaction between FtsEX and a single specific PG hydrolase called PcsB making it an attractive extracellular target to prevent pneumococcal disease. In this proposal we directly address questions concerning a particular essential enzyme and the complex it makes with cell division proteins in Streptococcus pneumoniae. The cell division proteins FtsE and FtsX form a complex together that spans the bacterial membrane and anchors an extracellular enzyme called PcsB that is required for cell division. The binding and hydrolysis of ATP inside the cell by FtsE is transmitted through its membrane anchor partner protein FtsX and results in a major conformational shape change in PcsB outside the cell which controls its ability to cut the peptidoglycan layer and allow cell division. Building on new structural data and models, genetic constructs, biochemical data, assays and international collaboration, the goal of this proposal is to understand the role of PcsB in complex with FtsEX and elucidate the molecular events linking cell division with PG degradative enzymes required for growth and division in S. pneumoniae. Drugs or vaccines that interfere with this process could prevent division and could provide routes to new treatments for pneumococcal and related infectious disease. The research proposed in this grant proposal forms part of an international effort with colleagues in the US and Singapore to combat this problem. The scientific principles that we will reveal may also have application in other, related bacterial species. Our work leading to this application, provides computer simulations, microbiological tools, techniques and biochemical approaches that can now be applied to the key biological questions of how are these proteins controlled, how do they function and how we might interfere with this this process to provide future antimicrobial strategies for human or animal health.

Life is a journey. As we grow older, we change. Sometimes we respond in the spur of the moment. Occasionally, an event has long-lasting consequences in spite of any change in circumstance and shapes our outlook far into the future. This future flexibility, or a lack thereof, also applies to the traits like size and weight that influence our daily risk of death and our reproductive success. Some of these traits retain flexibility throughout life, whereas others can only change in a fixed early window. As humans, we are far more likely to shift weight gain trajectories before six months of age than when older. Any ability to flexibly adjust traits can boost survival chances in new or changing environments, but also provides the means to innovate and so express new combinations of traits. Flexibility as a means of innovation might promote the divergence of ancestral organisms into new species, but also might not because such flexibility would mean that species can already deal with whatever circumstances they encounter, which would in turn remove the pressure for any innovation to become hardwired into their DNA. The long timescales over which this hardwiring plays out complicates collection of data. We don't know whether future flexibility or a lack of it is more likely to catalyse change into new species. In this project, we will contribute this increasingly requested data and therefore provide the first evidence if a lifetime of flexibility, or a stubborn refusal to change, influences the emergence of new species. Planktonic foraminifera are single-celled organisms that live in vast numbers in all the world's oceans. While chemical analysis of their fossil remains has generated a remarkably continuous record of past climate change, each individual also retains a complete record of its size and shape at each stage along its journey through life. These growth stages can be revealed by state-of-the-art imaging technology, which has sparked a digital revolution in how biologists study life on Earth. To study evolution, we need to study differences among lots of individuals. We need to know how and why these differences change through time. This need to measure lots of individuals means that the current practise of a person pointing and clicking on a computer screen to identify distinct parts is too slow. Computer programmes that provide a faster, more repeatable and less biased way of identifying and analysing such parts are now available, completing the toolkit needed to build big databases. By bringing together lessons from diverse scientific disciplines, we propose to use the same fossil specimens to collate records of an individual's journey through life and the environment it experienced every step of the way, both of which were changing from day-to-day, millions of years ago. While the fossil record of planktonic foraminifera provides the necessary timespan and abundance, new computer programmes and imaging technology complete the toolkit jigsaw to investigate for the first time if certain parts of an individual's journey through life are more influential than others in determining the eventual evolutionary destinations of its species. Our unique, direct link between organism and environment lets us study the dynamic journey through life in the static death of the fossil record. The fundamental limitation to the current ways we study how new species emerge is the lack of repeated samples through time to follow the genesis of novel lifeforms, and explicitly targeting this limitation using state-of-the-art approaches from multiple scientific disciplines means we will deliver a breakthrough in attempts to answer one of the most fundamental of all biological questions: how do differences among individuals make differences among species?

The overarching goal of our research programme is to address aspects of the broad science challenge: "What are the basic constituents of matter and how do they interact?". In particular, by performing experiments primarily with electron and photon beams, we study questions such as "How do quarks and gluons form hadrons?", and by studying these basic, strongly-interacting building blocks we are able to tackle the question "What is the nature of nuclear matter?"

Multiple myeloma is the second most common form of blood cancer and is a cancer of plasma cells, a type of white blood cell, which normally help to fight infections through the production of antibodies. Patients with multiple myeloma develop complications including low blood counts (anaemia), kidney damage, and bone damage including fractures which can cause significant symptoms. Multiple myeloma progresses from asymptomatic precursor conditions, but most patients are diagnosed at the active, symptomatic stage. There are treatments for multiple myeloma, but it remains almost always incurable, and most patients will die because of their myeloma. In multiple myeloma there are changes in the genes of the cancerous cells which can give rise to different behaviours and outcomes with treatment. These genetic changes can include gains or losses of whole or parts of chromosomes (copy number abnormalities), movement of genes between chromosomes (translocations), and changes in the sequence of genes (mutations). We know that some of these genetic changes are linked with worse outcomes despite treatment (termed high-risk genetics). Chromosome 13 is often lost in multiple myeloma and is one of the most common genetic changes, seen in about half of patients at diagnosis. The presence of chromosome 13 loss is associated with progression from the asymptomatic precursor conditions into symptomatic disease. Whether chromosome 13 loss itself is linked with worse outcomes in multiple myeloma is not entirely clear but it is seen more commonly with other high-risk genetic features and due to this, overall patients who have this abnormality have inferior outcomes despite current treatments. However, the impact of these changes on the myeloma cells remains largely unknown and we need a better understanding as it could make an ideal candidate for targeted treatments. This project has two linked aims: 1. To characterise the effect of chromosome 13 loss in myeloma genetically I will perform gene sequencing in samples from patients with newly diagnosed multiple myeloma enrolled into a clinical trial, including those with and without loss of chromosome 13 to assess the impact of this abnormality on other chromosomes and genes as well as on RNA (the intermediate step between DNA and proteins within the cell). With this information, I aim to improve understanding of: 1.What happens to other genes and why this might happen 2.Why some genetic events are seen more commonly with one another 3.How chromosome 13 loss may impact on proteins (which are the drivers of behaviour) in myeloma cells 2. To characterise the effect of chromosome 13 loss in myeloma functionally In myeloma cells which grow in the laboratory (myeloma cell lines), which have the normal two copies of the chromosome, I will use a recently described gene editing technique to create cells which have lost one of these copies mimicking what we see in some patient samples with myeloma. This will enable these cells to be grown and compared with those cells with the normal two copies in functional experiments. The advantage of this approach is that this will be the only significant difference between these cells and therefore the specific effects of this abnormality can be studied. These functional studies will aim to understand the impact on: 1.How these cells grow 2.How these cells respond to current myeloma treatments 3.How this change specifically impacts proteins in myeloma cells 4.Whether any of these changes can be targeted with new treatments The overall aim therefore is to improve the outcomes of patients with multiple myeloma who have loss of chromosome 13 through improving understanding of this genetic event on their cancer and how we may be able to treat this in a targeted manner.

The overarching goal of our research programme is to address aspects of the broad science challenge: "What are the basic constituents of matter and how do they interact?". In particular, by performing experiments primarily with electron and photon beams, we study questions such as "How do quarks and gluons form hadrons?", and by studying these basic, strongly-interacting building blocks we are able to tackle the question "What is the nature of nuclear matter?"