Proteome mapping of the model fungal plant pathogen Stagonospora nodorum using LC-LC-MS/MS. Stagonospora nodorum is a fungus that causes leaf and glume blotch disease on wheat. This disease alone causes $55 million dollars in yield losses per annum in Australia. This project aims to identify the proteins produced by Stagonospora nodorum through the development of a new proteomics technique. Two clear benefits to the community resulting from this project will emerge. The first will be the expert ....Proteome mapping of the model fungal plant pathogen Stagonospora nodorum using LC-LC-MS/MS. Stagonospora nodorum is a fungus that causes leaf and glume blotch disease on wheat. This disease alone causes $55 million dollars in yield losses per annum in Australia. This project aims to identify the proteins produced by Stagonospora nodorum through the development of a new proteomics technique. Two clear benefits to the community resulting from this project will emerge. The first will be the expert training of a student in proteomics, a skill that is keenly sought. Secondly, the identification of these pathogen proteins will lead to new strategies to better control the disease and secure the supply of wheat.Read moreRead less
Tail-anchored membrane proteins: prediction, targeting, assembly and function. Using computer-based searches of genome sequence data, we now have a complete list of tail-anchored membrane proteins in the yeast Saccharomyces cerevisiae. These include a number of essential proteins, such as SNAREs and TOM proteins responsible for building cellular membranes in all organisms, including man. Of the additional protein sequences discovered in the search, 8 represent proteins of known function while 19 ....Tail-anchored membrane proteins: prediction, targeting, assembly and function. Using computer-based searches of genome sequence data, we now have a complete list of tail-anchored membrane proteins in the yeast Saccharomyces cerevisiae. These include a number of essential proteins, such as SNAREs and TOM proteins responsible for building cellular membranes in all organisms, including man. Of the additional protein sequences discovered in the search, 8 represent proteins of known function while 19 are novel. We propose to study the subcellular location of these 19 novel proteins, and solve how they are targeted to and inserted in membranes. We will also investigate the function of the newly-discovered proteins.Read moreRead less
The Fine Tuned Physiology of Microaerophilic Gastric Spirilla. The aim of the project is to understand the molecular basis of fundamental properties of the physiology of enterogastric spiral bacteria of the genera Campylobacter and Helicobacter. The characteristics of these obligate microaerophiles which will be investigated are their aerobic respiratory chains, the special metabolites and enzymes involved in thiol-disulphide redox balance, and their essential requirement for carbon dioxide. Mic ....The Fine Tuned Physiology of Microaerophilic Gastric Spirilla. The aim of the project is to understand the molecular basis of fundamental properties of the physiology of enterogastric spiral bacteria of the genera Campylobacter and Helicobacter. The characteristics of these obligate microaerophiles which will be investigated are their aerobic respiratory chains, the special metabolites and enzymes involved in thiol-disulphide redox balance, and their essential requirement for carbon dioxide. Microaerobes include some bacteria, archea and protozoa. Realisation of the widespread habitats and importance of microaerophiles, has led recently to a vigorous interest in understanding their physiology. Knowledge of the basic properties of microaerophily has potential applications to Environmental Microbiology, Agriculture, Industrial Microbiology, Veterinary Science and Medicine.Read moreRead less
Genetic analysis of cohesin function and regulation in Drosophila. In yeast, a multiprotein complex, called cohesin, holds newly replicated chromatids together until the cell is ready to partition each chromatid into its daughter cells. We and others have shown that cohesins are regulated differently in animal cells. We propose to combine classical genetic analyses with two new and innovative techniques, time-lapse confocal microscopy of fluorescent proteins in living cells and gene-specific kno ....Genetic analysis of cohesin function and regulation in Drosophila. In yeast, a multiprotein complex, called cohesin, holds newly replicated chromatids together until the cell is ready to partition each chromatid into its daughter cells. We and others have shown that cohesins are regulated differently in animal cells. We propose to combine classical genetic analyses with two new and innovative techniques, time-lapse confocal microscopy of fluorescent proteins in living cells and gene-specific knockout techniques to study key cohesin regulators in Drosophila. These studies will provide us with novel insights into how multicellular organisms regulate the structure and stability of their chromosomes.Read moreRead less
A proteomic approach to identifying the signaling pathway(s) by which acute oxidative stress causes cell death by apoptosis. Oxidative stress following traumatic injury (heart attack or stroke) is known to activate signaling pathways leading to programmed cell death (apoptosis). The aim of this project is to develop methods to identify the signaling proteins involved. Identifying proteins involved in causing cell death will be useful in developing diagnostic tools as well as providing potential ....A proteomic approach to identifying the signaling pathway(s) by which acute oxidative stress causes cell death by apoptosis. Oxidative stress following traumatic injury (heart attack or stroke) is known to activate signaling pathways leading to programmed cell death (apoptosis). The aim of this project is to develop methods to identify the signaling proteins involved. Identifying proteins involved in causing cell death will be useful in developing diagnostic tools as well as providing potential therapeutic possibilities.Read moreRead less
The development of mass spectrometry techniques for mapping post-translational modifications in the wheat pathogen Stagonospora nodorum. The fungus Stagonospora nodorum is a significant pathogen of wheat causing in excess of $100 million dollars in yield losses per annum in Australia. This project will develop new analytical methods that can be used to detect important protein modifications in Stagonospora nodorum with the goal of securing Australia's wheat supply.
A Unique Target in the Purine Biosynthesis of the Pathogen Helicobacter pylori. The uptake systems of purine and analogues of the human pathogen Helicobacter pylori will be characterised because they can be utilised to introduce cytotoxic compounds into the cells. The first step in de novo purine biosynthesis of the bacterium is catalysed by two different enzymes, which are components of other biosynthetic pathways. These unique properties make them excellent potential therapeutic targets. Their ....A Unique Target in the Purine Biosynthesis of the Pathogen Helicobacter pylori. The uptake systems of purine and analogues of the human pathogen Helicobacter pylori will be characterised because they can be utilised to introduce cytotoxic compounds into the cells. The first step in de novo purine biosynthesis of the bacterium is catalysed by two different enzymes, which are components of other biosynthetic pathways. These unique properties make them excellent potential therapeutic targets. Their individual combined activities in purine biosynthesis will be characterised in situ and in vitro. Isogenic mutants with inactivated genes encoding for these enzymes will be constructed to investigate their role in the survival of the organism.Read moreRead less
Structure and function of the protein translocation channels in the mitochondrial outer membrane. Biological membranes are the basis of life and understanding how proteins are inserted into membranes is a major goal in the Biological Sciences. The TOM complex is a molecular machine mediating protein insertion into a biological membrane. Recent successes with X-ray diffraction of protein crystals, and solution and solid-state NMR spectroscopy have heralded leaps-and-bound advances for our knowled ....Structure and function of the protein translocation channels in the mitochondrial outer membrane. Biological membranes are the basis of life and understanding how proteins are inserted into membranes is a major goal in the Biological Sciences. The TOM complex is a molecular machine mediating protein insertion into a biological membrane. Recent successes with X-ray diffraction of protein crystals, and solution and solid-state NMR spectroscopy have heralded leaps-and-bound advances for our knowledge of how membranes work at the molecular level. Using a combination of phylogenetic analysis, yeast genetics and these new techniques from structural biology, we will characterize the structure and function of the core from the TOM complex.Read moreRead less
Investigating the molecular function of alpha-Haemoglobin stabilising protein. The research described in this proposal will provide new insights into haemoglobin regulation and redox chemistry in erythrocytes. Deregulation of these processes gives rise to a number of debilitating diseases, including varieties of anaemia and thalassaemia-in Australia it is estimated that 3% of the population could be carriers of b-thalassaemia mutations. Given the contribution of free aHb to the pathology of b-th ....Investigating the molecular function of alpha-Haemoglobin stabilising protein. The research described in this proposal will provide new insights into haemoglobin regulation and redox chemistry in erythrocytes. Deregulation of these processes gives rise to a number of debilitating diseases, including varieties of anaemia and thalassaemia-in Australia it is estimated that 3% of the population could be carriers of b-thalassaemia mutations. Given the contribution of free aHb to the pathology of b-thalassaemia, understanding the specific aHb-binding factor, AHSP is a goal of national significance. In the long term, manipulation of AHSP function through gene therapy may have a direct role in the treatment of thalassaemia.Read moreRead less
Force-from-lipids biophysical principle underlying mechanotransduction. The major aim of this project is to determine evolutionary conserved physical principles of mechanotransduction in living cells through structure and function studies of PIEZO mechanoreceptor channels playing a crucial role in senses such as touch and pain in animals and humans. Mutations in these channels can cause numerous genetic disorders, including hereditary anaemias and joint contractures. Since they have been shown t ....Force-from-lipids biophysical principle underlying mechanotransduction. The major aim of this project is to determine evolutionary conserved physical principles of mechanotransduction in living cells through structure and function studies of PIEZO mechanoreceptor channels playing a crucial role in senses such as touch and pain in animals and humans. Mutations in these channels can cause numerous genetic disorders, including hereditary anaemias and joint contractures. Since they have been shown to respond to mechanical stimuli in the same manner as mechanoreceptor channels of organisms from bacteria to humans the intended outcome of this project is to uncover the unifying principles of mechanotransduction anchored in the laws of physics and chemistry that have guided the force-dependent design of all life forms.Read moreRead less