E-cadherin is one of the major proteins responsible for mediating cell-to-cell adhesion in the body. During development, E-cadherin is essential for establishing the cellular architecture of epithelial organs and for maintaining epithelial function in the adult. In this context, E-cadherin acts to establish and maintain the polarity of epithelial cells. E-cadherin is also a powerful tumour suppressor and the loss of E-cadherin expression or function is a primary event in metastasis and cancer in ....E-cadherin is one of the major proteins responsible for mediating cell-to-cell adhesion in the body. During development, E-cadherin is essential for establishing the cellular architecture of epithelial organs and for maintaining epithelial function in the adult. In this context, E-cadherin acts to establish and maintain the polarity of epithelial cells. E-cadherin is also a powerful tumour suppressor and the loss of E-cadherin expression or function is a primary event in metastasis and cancer invasion. Proteins at the surface of epithelial cells must be sorted and trafficked, or transported, to different membrane domains. E-cadherin, for instance, must be trafficked to the lateral domain of cells in order to function in cell-cell adhesion. We recently discovered that cell surface E-cadherin is re-internalized and recycled back to the surface via a pathway that is poised to contribute to the regulation of cell adhesion. Our proposed studies aim to reveal how newly-synthesized E-cadherin and recycling E-cadherin are trafficked, which molecules and which vesicle carriers accomplish this transport. E-cadherin has specific amino acids that act as targeting signals for its sorting and trafficking; we have recently identified one such signal and will now seek the signal responsible for its endocytosis. Using specifically engineered mutants of E-cadherin we will also study other proteins that interact with E-cadherin during its trafficking for sorting and regulation. One of these is polycystin, a protein that is mutated in a common inherited kidney disease. Insights into this disease and normal kidney epithelial function will emerge from this work. A growing understanding of E-cadherin function and regulation is essential for the health of epithelial organs and for controlling and preventing cancer.Read moreRead less
I am a molecular and cell biologist with a major research focus on haemopoiesis and leukaemia development. This work principally involves the analysis of mutant mouse models.
Engineered Histones As DNA Carriers With Application In Therapeutic Gene Delivery
Funder
National Health and Medical Research Council
Funding Amount
$417,750.00
Summary
We intend to apply our knowledge of protein transport to the nucleus to enhance the delivery of DNA to target cells. This relates to the use of gene therapy to treat genetic defects such as inborn errors of metabolism, where a disease-causing lack-of-function mutation can be overcome by engineering cells within the organism which express, in the necessary quantities and in response to the appropriate regulatory signals, the particular component which is lacking. A limiting factor in gene therapy ....We intend to apply our knowledge of protein transport to the nucleus to enhance the delivery of DNA to target cells. This relates to the use of gene therapy to treat genetic defects such as inborn errors of metabolism, where a disease-causing lack-of-function mutation can be overcome by engineering cells within the organism which express, in the necessary quantities and in response to the appropriate regulatory signals, the particular component which is lacking. A limiting factor in gene therapy approaches is the low efficiency of nuclear uptake of introduced DNA, where it has been estimated that < 1% of the DNA taken up is actually expressed. Our proposal seeks to develop approaches to enhance non-viral-mediated gene delivery, in particular by optimising this critical, limiting step of the delivery of exogenous DNA to the nucleus. We intend to apply knowledge from studies of nuclear targeting and chromatin assembly to improve gene transfer technologies. We will build on our work showing that specific signals for nuclear import - nuclear targeting signals (NTSs) - can be used to enhance nuclear gene delivery and expression. Since DNA in the normal cellular context is in the form of chromatin - a specific complex with proteins such as histones - we intend to use reconstituted chromatin as the transfecting DNA, whereby histones engineered to include NTSs and other modular sequence elements will be used. Chromatin should not only enable NTSs and other sequence modules to be linked to the DNA but also protect against nuclease-mediated degradation prior to nuclear entry, condense the DNA to enable more efficient cellular-nuclear entry, and ensure expression of the transfected reporter gene by presenting it to the cell in a physiological context. Our approaches should contribute to bringing gene therapy closer to reality in the clinic.Read moreRead less
Molecular Mechanisms of Biochemical Regulation: Neutron and X-ray Scattering Studies. This project will develop and use novel neutron and x-ray scattering methods to study the molecular mechanisms by which nature regulates biochemical processes. Healthy function requires cells to tightly control and coordinate a myriad of molecular activities. My research focuses on a set of interdependent molecular networks inside cells whose behavior is controlled by the so-called 'second messengers' that tr ....Molecular Mechanisms of Biochemical Regulation: Neutron and X-ray Scattering Studies. This project will develop and use novel neutron and x-ray scattering methods to study the molecular mechanisms by which nature regulates biochemical processes. Healthy function requires cells to tightly control and coordinate a myriad of molecular activities. My research focuses on a set of interdependent molecular networks inside cells whose behavior is controlled by the so-called 'second messengers' that translate external signals into the right cellular responses. The proposed experiments will provide a unique structural framework by which we can understand how these signals are transmitted. Such knowledge is an important foundation for advances in biomedical research and biotechnology applications.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
Molecular mechanisms of two-component signal transduction in bacteria. The focus of this research is on the protein complexes that transmit signals in bacteria to elicit the desired responses to environmental stimuli. Like many dynamic processes in cells, signaling requires proteins that are flexible and hence resistant to high-resolution structural analysis using crystallography. We will make use of new research infrastructure at the Australian synchrotron and OPAL research reactor to overcom ....Molecular mechanisms of two-component signal transduction in bacteria. The focus of this research is on the protein complexes that transmit signals in bacteria to elicit the desired responses to environmental stimuli. Like many dynamic processes in cells, signaling requires proteins that are flexible and hence resistant to high-resolution structural analysis using crystallography. We will make use of new research infrastructure at the Australian synchrotron and OPAL research reactor to overcome the challenges of flexibility in these systems. The proteins we will study are not found in humans, and hence our research will provide important structural data on potential targets for the design of novel antibiotics to fight bacterial infection.Read moreRead less
Olfactory signal transduction in Drosophila melanogaster. This project will strengthen Australia's research capabilities in the areas of molecular neurobiology and neurogenetics. The project will equip students with the intellectual and technical skills needed to work in priority areas such as genomics and biotechnology, as well as in medical and agricultural research, and education. The research has possible long term applications in modifying the behaviour of insects of agricultural or medical ....Olfactory signal transduction in Drosophila melanogaster. This project will strengthen Australia's research capabilities in the areas of molecular neurobiology and neurogenetics. The project will equip students with the intellectual and technical skills needed to work in priority areas such as genomics and biotechnology, as well as in medical and agricultural research, and education. The research has possible long term applications in modifying the behaviour of insects of agricultural or medical importance. For example, by inhibiting the ability of insects to perceive specific odours it may ultimately be possible to prevent insects that carry disease from identifying target animals, or plant pests from locating their host plants.Read moreRead less
Olfactory signalling and coding in Drosophila and other insects. Animals rely on olfactory cues to detect food, danger, and others of the same species. The olfactory systems of Drosophila and other insects are simpler than those of mammals, yet complex enough to offer fascinating systems for studying neural information processing. This project aims to investigate the role of the Drosophila odorant receptors in olfactory coding, and to use multiple approaches to isolate components of the poorly u ....Olfactory signalling and coding in Drosophila and other insects. Animals rely on olfactory cues to detect food, danger, and others of the same species. The olfactory systems of Drosophila and other insects are simpler than those of mammals, yet complex enough to offer fascinating systems for studying neural information processing. This project aims to investigate the role of the Drosophila odorant receptors in olfactory coding, and to use multiple approaches to isolate components of the poorly understood insect olfactory signal transduction pathway. In addition, the role of the NO/cGMP pathway in olfactory signalling will be studied using an olfactory mutant that has nitric oxide synthase defects.Read moreRead less
Biogenesis of secretory organelles and the function of adhesins secreted during the establishment of plant disease. Many agriculturally important crops and Australian native plants are susceptible to diseases caused by species of Phytophthora, a fungus-like organism that lives in the soil. Economic losses due to Phytophthora diseases are estimated to exceed $200 million per annum and the scale of environmental damage in natural ecosystems is huge. Currently, control of Phytophthora diseases la ....Biogenesis of secretory organelles and the function of adhesins secreted during the establishment of plant disease. Many agriculturally important crops and Australian native plants are susceptible to diseases caused by species of Phytophthora, a fungus-like organism that lives in the soil. Economic losses due to Phytophthora diseases are estimated to exceed $200 million per annum and the scale of environmental damage in natural ecosystems is huge. Currently, control of Phytophthora diseases largely depends on a very small number of effective chemicals and there is an imminent risk of the development of pathogen resistance. This research will increase our understanding of how Phytophthora spores infect host plants and will identify suitable targets for the development of novel, environmentally safe chemicals that inhibit disease development.Read moreRead less