Directors’ report / 2 IMB seminars / 78 Highlights 2000 / 6 Collaborations / 81 Group profiles / 9 Financial / 82 Joint ventures / 72 Publications / 84 Graduate program / 75 Staff list / 90

IMBcom / 76 Contents

Genomics & Bioinformatics 9 & Developmental17 Biology 33 Structural Biology49 Biological Chemistry57 IMB Associates68 2 3 imb/2000

Professors John Mattick and Peter Andrews, Co-Directors of the Institute for Molecular Bioscience, talk with Peter McCutcheon

Why is Queensland, which has Professor Andrews: The world’s most including industry. The IMB hosts traditionally relied on mining, rapidly growing economies are all the ARC Special Research Centre for agricultural and tourism industries, knowledge-based industries, and the Functional and Applied Genomics and developing an advanced research focus of those industries is increasingly is a core partner in the Cooperative centre for molecular bioscience? on biotechnology. In fact, recent Research Centre for the Discovery of advances in genetic engineering Genes for Common Human Diseases, as Professor Mattick: It is clear that we are techniques and the sequencing of the well as being the host (along with the in the midst of one of the greatest periods human genome have extended the Walter & Eliza Hall Institute of Medical of discovery in history – the genetic and applications of biotechnology from Research) of a major national research molecular basis of life and its diversity. common foodstuffs into most aspects of facility, the Australian Genome Research The genome projects are laying out the our lives, including , energy and Facility. So we do have a good balanced genetic blueprints of different species, the environment. For me, the question is portfolio of research funding. from microorganisms to humans, and not why would you do it in Queensland, functional studies are leading us to it’s why wouldn’t you! In terms of whether we’ve got enough, no understand how these components work we don’t. Research funding in Australia together to produce complex outcomes. The Queensland Government has made remains low by international standards, And the flip-side of this is to understand a significant investment in the Institute. and our ability to attack big problems and disease. Sooner or later this knowledge Are there other funding arrangements create new opportunities is limited by the and its applications will transform all of in place for the IMB, and is it enough? relatively restricted resources available the biologically-based industries, which to us. Our funding is still well below occupy perhaps half the world economy, Professor Mattick: The Queensland that of comparable Institutes in other and lead to entirely new industries based Government has been generous to the parts of the world including Asia. We on understanding genetic programming IMB, and has vested a great deal of need more funds to mount strategic genomics projects, to build the computing infrastructure for bioinformatics, to “Queensland is a state with great dynamism and enable the next generation of structural analysis in cell biology, and to develop we are very fortunate to have a government that new chemical libraries, to mention a few. understands the importance of developing an Although we are doing the best we can, advanced, knowledge-based economy.” and will do well with what we have. My view, shared by many others, is that there is insufficient attention paid in our and biological chemistry. So this faith in us, which we will work hard society to investing in the future, which is clearly a fundamental knowledge to justify and repay. We have also is what research is. It’s a bit like a revolution that has huge potential and been fortunate to have raised funding home budget – you have to pay the will have huge ramifications. from the Federal Government through bills, buy the food, maintain the house. the Federation Fund, and from a private But the amount of money you can save Why Queensland ? – I think that donor, to construct the Institute. And and invest for the future determines your Queensland is emerging as the California we’re delighted to have the CSIRO now future prosperity and quality of life. of Australia. It not only has great joining us in the construction of the It’s the same for the nation as it is natural resources and primary industries Institute complex, a joint venture that will for a household. So I’d like to see but has great capacity to develop new create a very large and vibrant centre for more emphasis on this aspect of national high technology industries, and this is advanced research in development, and an acknowledgment already underway. It is well positioned and biotechnology. that as much of our natural resources as in the Asia-Pacific region. Queensland is possible be invested on everyone’s behalf also a state with great dynamism and we The IMB also obtains substantial funding to create better futures, especially in the are very fortunate to have a government for its research projects and programs areas of education, R & D and strategic that understands the importance of from the Australian Research Council, the industry development. Other countries in developing an advanced, knowledge- National Health and Medical Research our region have been doing this for the based economy. Council, the Cooperative Research past 20 years, and the benefits are very Centres scheme, and many others, obvious. 2 3

The IMB depends on private as well as are genetically engineered insulin and Queensland’s is one of two state public funding. Although the Australian erythropoietin (EPO). The market for governments which has announced private sector doesn’t have a strong these products last year was roughly heavy investment in biotechnology, the track record on R & D investment, US$9 billion - greater than the entire other state of course is Victoria. How is there any sign of a change in will Victoria’s initiative affect the IMB? Australian grains crop. investment culture?

The IMB is unusual in that it comprises Professor Mattick: We are very Professor Andrews: There’s a real breath both basic research facilities and spin- supportive of the Victorian initiative. of fresh air emerging in the Australian off companies. It has been described Victoria has traditionally been the venture capital community, and a lot of as a “pipeline” from genomics to strongest centre of biomedical research in them are looking to the IMB as a source pharmaceuticals. Why is it set up that this country. It is also now clear that of new investments. We’ve had three way? Brisbane has emerged as a major centre venture capital firms invest in spin-off in its own right, and will additionally companies from the IMB in the past Professor Mattick: Because we would become a major centre for advanced year, and we’re currently in negotiation like the IMB to fulfil two objectives. biological research and biotechnology. with several firms interested in IMB First and foremost, we want to earn This is important for Australia. We technology platforms that may form the a reputation as being a very fine have many partnerships with Victorian basis for further spin-offs in 2001. research institute, capable of winning research institutions in areas such as the Nobel prizes. New knowledge is New industries by definition can be unpredictable. The NASDAQ index dropped 35 percent in the US in 2000. Is biotechnology really becoming “New knowledge is not only important to a credible investment item for the understanding our world, it is also the basis for Australian business community? economic development, as the history of the Professor Mattick: I think it is. past two centuries shows.” You’ve got to see the stock exchange as partly a reflection of commercial and industrial reality, and partly a reflection of not only important to understanding Australian Genome Research Facility. So people making considered bets on future our world, it is also the basis for we see ourselves as friendly rivals but industrial development. In the short-term economic development, as the history of above all as partners. We are Australians the latter is unpredictable; in the longer the past two centuries shows. Secondly, first and we have to work together, we do term there is no doubt that, in relation therefore, we also want to fulfil our work together, to make the most of our to biotechnology, the direction is up, responsibility to those that invest in resources and research strengths. and the figures bear that out, although us, by earning a reputation as being market confidence waxes and wanes. I’ll well connected with industry, and having Although the IMB is large by Australian give you one example of the economic a very pro-active stance in terms of standards, it is modest by international standards, especially compared to value of this sector. Two pharmaceuticals taking our discoveries, technologies and developments in the United States. expertise into practical applications. This which are widely used in medicine Can the IMB be a serious player in in turn will feed back into support for – and interestingly there’s very little global science and biotechnology? public concern about these products – further research. Professor Andrews: The by-product of pharma industry mergers is a vast increase in the extent to which the industry outsources its R&D to research organisations and biotechnology companies. And that’s where we have a huge opportunity to be very serious players.

One fascinating example is a molecule discovered by IMB researchers in the venom of a marine cone snail from the Great Barrier Reef which has been licensed to Australian pharmaceutical company Amrad for clinical trials as a pain therapeutic. This and several other lead molecules with novel pharmacological activities are currently progressing through the pipeline of an IMB spin-off company, Xenome Ltd. 4 5 >>imb/2000

Another example is our work in because we are not necessarily being Professor Andrews, there has also designing molecules mimicking growth swept along with current fashions. I been some public debate over the hormones and other cytokines. These think that’s always been something of a patenting and selling of genetic are that send signals from hallmark of Australian science. Being out information, particularly in relation to the actions of the US company, Celera. one cell to another by interacting very of the mainstream has had its down-side, What is your view on the commercial selectively with other proteins on the cell but its up-side is that Australian scientists use of research findings? surface. These and other -protein tend to be a bit more creative and think interactions represent one of the biggest about things more laterally. Professor Andrews: The reality is that while it costs so much to create a new drug, companies need the protection that a patent gives them to effectively develop and market them. But if you ask “Should Celera be able to patent the human genome?” – that’s a lot more difficult to answer. I find it difficult to handle the concept of patenting gene sequences. “Australian The next stage on, when we can say scientists “Now we have the sequence of this gene and we know its function”, is a tend to be a much more defensible basis for patenting, bit more and when it comes to developing a new product based on our knowledge of creative and the gene and its function, patents are think about absolutely essential. things more The IMB is accommodating a wide variety of personnel from basic science laterally.” researchers, spin-off company employees through to patent lawyers. Will this lead to major changes in research culture?

Professor Andrews: Well it is already leading to a change in culture. I think scientists throughout the IMB are increasingly recognising that if they are to see something valuable, in a public classes of potential therapeutic targets, A particular strength of the IMB is good sense, emerge from their research, but no one has been able to develop our integration of genomics with they also need to be thinking about how generic ways of attacking them. Dr Mark chemistry and our heavy emphasis on to develop it. So there’s a lot more Smythe’s group, working with Glaxo the emerging fields of bioinformatics interest in intellectual property, and how Wellcome and the National Health and and . Biology to protect it, as well as working with Medical Research Council (NHMRC), is becoming an information science industry. has solved that problem, and that gives us in its own right, with an increasing a very serious edge in a major aspect of intersection with information technology. The other big cultural change is in drug development. We are positioning the Institute to be the way our scientists think about their an international centre of excellence career options. In the past, postgraduate These examples are interesting – Xenome in this, and we have developed close students and postdoctoral fellows in because it utilises Australia’s natural relationships with some of the key research institutions had little option but resource strengths as well as our science, players internationally. to pursue limited career opportunities and the cytokine project because it in universities or other public research illustrates that a bunch of scientists sitting Also now with the Internet and improved organisations. Now, a good percentage in an Institute like this a long way from communications and travel, Australia is of our graduates and postdocs are finding the main game can nevertheless be the much better connected with the rest of great career prospects in start-ups and best in the world. the world. So it doesn’t matter so much spin-offs associated with IMB. whether something is in Brisbane or What do you see as the strength of the Tokyo or Paris, and in fact we collaborate Professor Mattick, you mentioned earlier there has been very little public IMB in an international context? with laboratories all around the world. concern about genetically modified We’re part of a network and part of the . They are widely used. Professor Mattick: One strength is that, feature of the establishment and growth However in parts of Europe the debate being somewhat separated from the rest of this Institute is the development of over genetically modified foods has of the world, we can be more creative partnerships internally and externally. extended to pharmaceuticals as well. 4 5

develop the concept of the IMB and to bring it to fruition. We freely agreed to amalgamate our two research centres (the Centre for Molecular and Cellular Biology, and the Centre for Drug Design and Development) to form the core of the IMB. I enjoy Peter’s company, both professionally and personally, and appreciate his advice and wisdom. We work extremely well together and have complementary skills and experience, which I think adds another dimension to the Institute’s potential.

Professor Andrews: Of course, the ultimate relationship is still evolving. Over the past year we’ve played dual “A bunch of scientists sitting roles as ex-directors of our original research Centres and as Co-directors in an institute like this a long of the Institute. In the latter role, way from the main game can John is increasingly picking up research responsibility across the entire spectrum nevertherless be the best in of our activities, and I am taking on much the world.” more of the commercialisation activities. The bottom line is – we’re having fun!

Professor Mattick: I would also add that we have both enjoyed the support and involvement of other key players in the development of the Institute, such as Professor Kevin Burrage who is the How do you see the debate playing out Every time that there is a major Director of the Advanced Computational in Australia? technological change, there is a section Modelling Centre and is helping to of society who are inclined to point out develop our bioinformatics capabilities, Professor Mattick: In the end, any some of the problems, real or imagined, as well as the Deans and other colleagues applications of modern biotechnology which may arise from the technology. in the Faculties of Biological & Chemical are only going to be taken up in the My personal view is that you keep a level Sciences, Health Sciences, Engineering, marketplace if they are perceived as head. You also have to progress. It is Physical Sciences & Architecture, and being useful and safe by the people who interesting for scientists to learn about Business, Economics & Law, and other use them – that’s the public. By and how life works, to understand ourselves, organisations such as CSIRO, the large there has been wide acceptance of the natural world and the basis of disease. Queensland Department of Primary medicines produced from biotechnology Out of that will come some sensible Industries, and the Queensland Institute because the people who need those applications and the public will respond of Medical Research, who are medicines know that they are useful, safe according to their own priorities and collaborating closely with us through and well-tested. needs. joint appointments and through the development of joint research programs The benefits of genetically modified food It’s unusual for an Institute like this to and facilities. The strong support of are not so obvious yet to the consumer, be run by two Co-directors. How is it the Vice-Chancellor, Professor John Hay, working out? although they are more obvious to the Deputy Vice-Chancellor Research, producers because they lower insecticide Professor Paul Greenfield, and other Professor Mattick: It is working out loads, increase yields and will eventually senior colleagues at the University of very well. Peter and I have worked allow more innovative and higher quality Queensland has been essential throughout closely together for some years now to food products. But in the end it’s and is much appreciated. not a matter of saying you should or shouldn’t buy this. People will decide for themselves depending on what they see as the benefits to them, and I am quite confident that good outcomes will occur, despite some overstated claims by Peter McCutcheon is the Assistant Director of the University’s Office of Marketing and the proponents and fear mongering by the Communications. He has worked for the ABC in Sydney, Darwin, Melbourne and most ideological opponents of biotechnology. recently in Tokyo as the ABC’s North Asia Correspondent. 6 7

y2k high lights

^ Ångstrom Art is a new initiative for the IMB and is the brainchild of a group passionate about visual art and committed to raising the awareness of science in the community. Their idea is to use stunning images sourced from current research and to link these images to a website providing background information about the science behind the image.

Ångstrom Art was awarded a 2000 Science and Technology Awareness Program grant of $50,000, to fund the distribution of a series of science image postcards throughout Australia. Keep an eye out for these Ångstrom Art postcards, coming to a café near you in 2001.

In November, Ångstrom Art hosted the “Art in Science” competition that attracted over 30 entries from within IMB. Images were judged by a panel that included UQ dignitaries and a Brisbane artist. One of the winning entries, a cellular Christmas tree by Kylie Georgas, was chosen for the IMB Christmas card. Other winners were “Fluoro Macrophage Dragon” by Wenda Shurety and Darren Brown and “Chromosome” by Alisa Poh (illustrated). An image by Michael Dooley of a molecular Christmas wreath was subsequently selected for the University of Queensland Christmas card.

Ångstrom Art is looking to expand its activities and include scientific art from outside IMB. Further details can be found on the Ångstrom Art website at www.angstrom-art.com. 6 7

Cardiovascular breakthrough >

Drs Peter Koopman and George Muscat discovered a gene that could lead to breakthrough treatment for many of the cancers that kill 34,000 Australians yearly.

Sox18 is the gene responsible for the development of blood vessels and hair follicles, and this gene has the potential to significantly effect the growth of tumour cells by restricting the growth of blood vessels that supply nutrients and oxygen, effectively starving a tumour.

Sox18 could also have applications in speeding up blood vessel development to help heal wounds faster and treat circulation problems common that lead to amputation in diseases like diabetes.

Actual treatments could still be several years away but this is a significant discovery in the understanding of human development and the application of this understanding to human health care.

This work was published in the prestigious science journal Nature Genetics and is the product of six years work. y2k high lights

Frontier women In September The Courier Mail published a feature article highlight- ing the achievements of IMB Group Leaders (L-R) Jenny Martin, Judy Halliday, Melissa Little, Carol Wicking and Jenny Stow . The story dealt with issues such as the speed of scientific advancement, returning to Australia to do scientific research and how science can become a lifestyle rather than just a career. 8 9 in brief y2k highlights... Top-down approach The IMB was visited in September by Skin cancer test the Hon. Kim Beazley, Leader of the Federal Opposition. IMB Group Leader Dr Rick Sturm and his research team, in collaboration with Professor Nick Martin from The Queensland Institute of Medical Research, have developed a genetic Mr Beazley was interested in the test that could help identify an individual’s susceptibility to skin cancer. impact biotechnology will have in the coming years and where the IMB fits The procedure, published in the American Journal of Human Genetics, identifies any into these coming developments. variations of the gene called Melanocortin-1 or MC1R. This gene confers red hair colour and the ability to tan. With excessive exposure to the sun every person can develop skin cancer, however the Achievement Award danger is higher for people with a variant of the MC1R gene. The new test, in conjunction John Mattick was presented with with appropriate counselling, could drastically reduce the incidence of skin cancers in The Eppendorf Achievement Award, Australia. recognising his scientific research contributions and his promotion of Sixty percent of Queenslanders have sunspots or lesions removed in their lifetime and 250 molecular biology throughout people in Queensland die from skin cancer each year. Australia.

Cash instalment IMB researchers recognised The first allocation of $3.75 million for the IMB was Several IMB senior scientists were handed over to the University as part of the State successful in the 2000 round of Government’s ten year, $77.5 million commitment to promotions. Peter Koopman was partly fund the operational costs of the Institute. promoted to Professorial Research IMB Co-director Professor Peter Andrews said the Fellow, while Melissa Little and Mark funding allocation is an important step towards build- Smythe were both promoted to ing a vibrant new knowledge based economy. Principal Research Fellows. George Muscat became a Principal Research “The IMB brings together Research and Development, Fellow of the NHMRC. commercialisation, Queensland government funding commitments and spin-off companies that will herald the beginning of an exciting new era in Queensland development,” said Professor Andrews. Four new research groups in IMB The year 2000 saw the establishment of four new research groups in the Asian network IMB. Rohan Teasdale, Mark Ragan, Jennifer Hallinan and Sean Grimmond The IMB has been elected as the 9th Centre of Excellence in the bring their extensive experience in bio- Asia-Pacific International Molecular Biology Network (IMBN), which is informatics, genomics, computational being established as a regional organization for research co-operation and biology and microarray analysis. training in molecular biology, similar to the European Molecular Biology Organization (EMBO). This places the IMB alongside other prestigious Institutes from the region, including the Weizmann Institute of Science; Olympic torch bearer IMCB, Singapore; Institute of Medical Science, University of Tokyo; the IMB group leader Judy Halliday had Walter and Eliza Hall Institute, Melbourne; and the Institute of Biological the honour of participating in the Chemistry, Academia Sinica, Taiwan. Olympic torch relay on June 14. AMP nominated Judy as a result of Boost for Queensland crystallographers her work with the Australian Society for Medical Research and AMP, in Biotechnology in Queensland was given a major boost with the announcement of promoting the AMP-QLD Biomedical substantial funding for a world class protein crystallography facility. Research Awards. The Australian Research Council awarded the University of Queensland $1.1 million for the facility. This grant is the largest individual national grant in the ARC’s Research Infrastructure Equipment and Facilities (RIEF) round. IMB Associate Professor Jenny Martin, until recently head of the only protein crystallography research group in Queensland, said the relocation of several key scientists to Queensland and the rate of growth of biotechnology initiatives in this state provided the impetus for this grant’s success. The facility will allow the visualisation of the largest and most complex biological molecules and this information will be critical in the design of novel and improved therapeutics. 8 9 Genomics Bioinformatics &

The interface between biology and computer science promises a more quantitative understanding of the complexities of living organisms, and new models for advanced computation. We apply computer-based methods to problems in molecular biology, and develop new approaches and software to investigate complex biochemical and biophysical processes. 10 Program in Genomics & Bioinformatics 11

Pseudomonas aeruginosa genomics and pathogenesis

Pseudomonas Analysis of the P. aeruginosa on cyclic di-GMP, all of which are genome likely to play important roles in aeruginosa infection The complete sequence of the pathogenesis poses a serious threat genome of P. aeruginosa, the largest bacterial genome sequenced to date Genes encoding type IV to immuno- (6.2 Mb), was published in 2000. Our fimbriae compromised group was involved in the annotation Type IV fimbriae are long filaments patients, burns of this genome [see Nature 406, which emanate from the pole of victims and cystic fibrosis sufferers. This bacterium produces many virulence factors which are used to colonize host tissues. We are identifying the important genes and proteins involved in infection, and the regulatory pathways controlling their expression, using a combination of genetic, genomic and 959-64 (2000)] as well as in more the bacterial cell and which impart biochemical detailed analyses which revealed a a form of surface translocation large number of unexpected genes called twitching motility. We have approaches. encoding, among others, four completed a high density transposon complete chemosensory systems, mutagenesis analysis of the P. three type II protein secretion aeruginosa genome and identified pathways, a number of surface over 40 genes involved in the haemaglutinin-like adhesins, three biogenesis, function and regulation of types of type I fimbriae, and a these structures. Many of these genes new global physiological regulation are homologous to genes involved in / signal transduction system based type II protein secretion and DNA 10 Program in Genomics & Bioinformatics 11

genomics and pathogenesis Group leader • John Mattick Assistant group leader • Cynthia Whitchurch

Postdoctoral staff • Ben Huang

uptake, revealing that each of these Research assistants • is an evolutionarily and structurally Jacqueline Emery, Jen Sargent related subset of a supersystem which places multimeric complexes on the Students • Scott Beatson, cell surface. Larry Croft, Andrew Leech, Michael Young Regulation of virulence factors We have identified a number of signal transduction systems which regulate the expression and function of type IV fimbriae, including PILSR, FimS/ AlgR, Vfr (virulence factor regulator) and a complex chemosensory system whose central component ChpA is the most sophisticated signal transduction protein yet described in nature. The last three of these systems also control other virulence factors, such as secreted proteases, toxins, pyocyanine and lactones, and appear to be components of a global regulatory system that intersects via phosphotransfer cascades and connects environmental signals with molecular motors and . 12 13

The human genome contains around 30,000 genes, but the vast majority of the output of the genome is non-protein-coding . We are examining the hypothesis that noncoding RNAs are not junk, but comprise a higher order parallel processing system underlying sophisticated genetic programming in the higher organisms. RNA regulation

The role of and other conserved from yeast noncoding RNAs (Saccharomyces cerevisiae) to Introns and other noncoding RNAs human, with homologues in mouse, comprise the vast majority of Drosophila melanogaster, genomic output in the higher Caenorhabditis elegans, Arabidopsis . Where studied these thaliana, Schizosaccharomyces transcripts have been found to be pombe, and other species. Nrap is developmentally regulated and to an essential protein and a core have genetic function. Analysis component of eukaryotic cell biology of complex genetic phenomena, which appears to interact directly or including co-suppression, transgene indirectly with the 45S rRNA. silencing, RNA interference, imprinting, and transvection, G3BP, a new signal Group leader • John Mattick indicates a far wider role for RNA in transduction protein gene control than has been previously We have discovered a new signal Assistant group leader • recognized. We are exploring the transduction protein G3BP which is Derek Kennedy possibility that RNA may have conserved in animals and links RNA evolved to function as a higher processing to the GAP120 signal Students • Larry Croft, Budi order parallel control system for the transduction pathway. In mammals Utama, Andrea Verhagen, Juliet integration and multitasking of gene there are two forms of this protein, French, Katherine Irvine, Emily networks in the higher organisms, and a number of splice variants McGhie analogous to internal control codes which appear to be developmentally in computer programming and regulated. The protein has a number Research assistant • Kelin efference signals that underlie motor of functional domains, including an Ru coordination and cognition in RNA recognition motif and nuclear neurobiology. import signals. We are investigating the role of this protein in normal Nrap, a new nucleolar protein cell biology and in cancer (where Nucleoli are membraneless nuclear its expression is disturbed) and organelles that are essential for attempting to identify its RNA and the and processing of protein interactions using a range of ribosomal RNA. We have identified biochemical and genetic approaches. a previously unknown nucleolar protein (Nrap, nucleolar RNA associated protein) that is highly 12 Program in Genomics & Bioinformatics 13

DNA microarray technology and yeast two-hybrid protein-protein interaction assays generate large amounts of complex biomolecular data. Our group is interested in using such datasets to build computational models of cells as networks of information flow.

Modeling of cell networks

Dynamics of cellular networks Naturally occurring networks that are large and complex tend to have a high degree of connectivity arranged so that the number of steps between individual points is vastly smaller than the size of the network. These are called small world networks, as in the "six degrees of separation" phenomenon. Such networks contain a few highly connected nodes, and many less highly connected nodes. Small world networks have characteristic dynamics, being exceptionally resistant to fragmentation by random node deletion, but vulnerable to targeted attack. Several metabolic networks have been identified as small world networks.

We have demonstrated that the network of protein-protein interactions in Saccharomyces cerevisiae forms a small world network, and exhibits the expected response to damage. However, metabolic networks are better modeled as directed graphs, and little is known about the implications of directed links between Group leader • Jennifer nodes for the behaviour of a small world network. We are currently building Hallinan a more comprehensive model of protein-protein interactions in S. cerevisiae, incorporating data such as protein subcellular location. This model will be used Students • James Tsai, to test theories of the evolution and dynamics of directed networks which we are Anthony Percival developing using simulated idealized small world networks.

GeNexus microarray database In conjunction with Sean Grimmond’s group we are designing and implementing

a relational database to hold the data generated by the IMB Microarray Facility. Once the database is completed we will be incorporating data mining and machine learning algorithms into the system to facilitate the identification of potentially interesting trends and patterns in the data. 14 Program in Genomics & Bioinformatics 15

Bioinformatic discovery of endosomal proteins

A major advance in Dissecting the endosomal involved in trafficking within these system through bioinformatics organelles, is to characterise human the biological The endosomal/lysosomal system of homologues of proteins that have sciences over the last mammalian cells is a highly dynamic been implicated in endosomal decade is the trafficking pathway that includes function in other organisms, membrane transport from the late predominantly yeast. This analysis sequencing of the Golgi and the plasma membrane. has identified over thirty novel genomes from The primary function of endosomes human proteins likely to function in is the sorting and segregation of the endosomal system. Specifically different organisms, receptors and ligands, a process we have focused on the sorting including the human that is necessary for many cellular nexin family of proteins and the genome. “Database operations. The molecular details of retromer subunits Vps26p, Vps29p protein trafficking and biogenesis and Vps35p. mining” allows for of the numerous subcompartments the prediction of the of the mammalian endosomal/ Protein trafficking in parasites lysosomal system are poorly defined. The Trypanosomatidae comprise a functional properties One strategy to identify proteins large group of parasitic protozoa, of a new protein based on the Sorting Nexins information contained within these genomes. The goal of my research is to extract this information so as to open up new avenues of scientific exploration within cell biology. 14 Program in Genomics & Bioinformatics 15

endosomal proteins Group leader • Rohan Teasdale

Students • Kevin Miranda, Rachael Shaw

Intracellular localisation signals A major issue in cell biology is how distinct intracellular regions of the cell maintain their unique composition of proteins and lipids. For the organelles to maintain their functional integrity, specific resident proteins must be retained while non- resident proteins are allowed passage through them. Individual proteins have “signals” that are responsible for their intracellular localisation. We are currently identifying such some of which cause important signals on several different proteins. diseases in humans. These include In addition, we are identifying novel Trypanosoma brucei (the causative sequences within the human genome agent of African sleeping sickness), that contain these localisation signals. T. cruzi (the causative agent of We have recently characterised a Chagas disease in Central and South family of proteins with a Golgi America) and Leishmania. Parasite Localisation Domain (GLD) and survival requires the surface identified the signal responsible for expression of specific molecules the correct targeting of E-cadherin to that form protective surface coats, the basolateral membrane. mediate specific host-parasite interactions and allow the parasite to take up essential nutrients. The biosynthesis, processing and surface transport of these molecules within the intracellular organelles of the parasite secretory pathway are poorly understood. A recent effort to sequence the genomes of these parasites provides us with an opportunity to identify parasite homologues of proteins that function in membrane trafficking. A collaboration with Dr. Malcolm McConville (University of Melbourne) has been established to characterise these numerous proteins. 16 17

We use advanced computer methods to investigate how genomes (and the proteins they encode) are similar to or different from each other, and to make quantitative inferences about how genomes have come to have their observed contents, arrangements and sequences of genes.

Comparative and computational genomics

The genealogical history of a and medicine. gene (or protein) can be inferred from its nucleotide (or amino In collaboration with Robert acid) sequence. Although histories Charlebois (NeuroGadgets Inc.), I (phylogenetic trees) inferred from continue to explore rapid different types of genes are often computational methods for identical, sometimes they are not, identifying atypical genes in even with methods that are robust microbial genomes. Some of these against methodological artifact. This atypical genes may be the products incongruence suggests that within of lateral transfer. However, other a single organism (ie within one evidence suggests that all “atypical” genome), different genes have genes can be of lateral origin. Group leader • Mark Ragan different genealogical histories. Many of these conflicts are difficult Thus I have begun to construct to explain unless we hypothesize an automated computer-based system lateral (horizontal) gene transfer: to infer and analyse phylogenetic that is, that the gene has entered trees from complete sets of genomic that genome not by “vertical” sequence data. Since arriving at IMB transmission from one generation to in mid-August, two UQ students the next, but by jumping, as it were, and I have implemented some basic from one branch of the phylogenetic modules of this system. We are tree to another. now refining many of its features. Although phylogenetic inference per Some analyses now suggest that se is a major goal of this fundamental lateral transfer may be very research, very similar approaches can common. If so, this could be and will be implemented to generate of both fundamental and practical databases optimised for mining and importance. The rapid spread of recovery of data useful in applied antibiotic resistance between areas of biosciences, including drug populations of bacteria, resulting design and metabolic engineering. in the emergence of drug-resistant “superbugs”, makes it clear that Research staff and students are some genes do move laterally. If, currently being recruited for this however, this is only the tip of core project, and to develop in-depth the iceberg, consequences could be projects in structure-based alignment, immense throughout environmental analysis of unique genes, and other science, biotechnology, agriculture related topics. 16 17 Genetics &

These are exciting times in biology. The availability of the genome sequences of humans and other organisms is revealing the genetic nuts and bolts of our makeup. Our research is providing spectacular insights into the functions of these genes, how they control our development and life, and how genetic malfunctions result in inherited disease. 18 Program in Genetics & Developmental Biology 19

Molecular genetics of organ devel opment

The development of Genetic control of sex organs critical for sexual identity and determination reproductive ability. complex organs in the A major focus of our group is embryo relies on the sex determination, the genetically Sox gene function in mouse co-ordinated controlled “decision” between male embryo development and female pathways of Study of the Y-linked testis- formation, migration, development, that manifests in the determining gene Sry has led us communication and embryo as a choice between testis to discover a number of related and ovary formation. We are genes known as Sox genes. These function of large studying several genes involved in are involved in a broad range numbers of cells of testis determination, such as Sry of processes such as skeletal, many different types. and Sox9, and continue to make cardiovascular, and neural significant progress in understanding development. Sox genes encode We are identifying the structure and regulation of transcription factors (regulators of genes involved in the these key genes. Using microarray gene activity) that specify the identity technology we have now discovered of a number of different cell types, development of the a large number of other genes active and are thus key players in directing gonads, skeleton, during testis or ovary development. organ development. Several of these blood vessels and Our goal is to piece together the genes are being studied in this molecular events that orchestrate the laboratory from a functional other organs. formation of testes and ovaries, standpoint. In addition, we have Studying how these genes function and interact provides a basis for understanding the complexities of embryonic development and how birth defects can arise. 18 Program in Genetics & Developmental Biology 19

organ devel opment Group leader • Peter Koopman

Postdoctoral staff • Josephine Bowles, Monica Bullejos, Jane Olsson, David Pennisi Sox8: A novel gene involved in Students • Kelly Loffler, embryo development Goslik Schepers, James Smith We have discovered a novel gene, Sox8, in humans and mice. Sox8 Research assistants • is extremely similar to Sox9, and, Vanessa Caig, Kristy James, is expressed in Sertoli cells of the Sarah Penning mouse fetal testis around the time of sex determination. We propose that Sox8 plays a role in male sexual development. Sox8 is also expressed in the developing brain and spinal cord in mouse embryos, in addition played a leading role in nomenclature to branchial arches and nascent facial and classification of these genes, structures. We mapped human SOX8 and studying their evolution and close to the telomere of chromosome functional relationships. 16, in a region associated with the facial malformation, mental Sox18: A critical gene in retardation and genital anomaly cardiovascular development syndrome ATR-16. SOX8 is therefore In collaboration with George a strong candidate for contributing to Muscat’s group, we have shown the features of ATR-16. that one Sox gene, Sox18, is active during the development of the blood vessel system and hair follicles in the embryo. The structure and expression of Sox18 is conserved among mice, humans and chickens. We found that mutations in Sox18 underlie a naturally-occurring mouse mutant that has defects in coat and vascular development, further emphasising the importance of this gene. Vascular development is clearly essential for survival of the embryo, and is also required for the growth of tumours. We are currently studying whether vascular development can be accelerated (eg. in wound healing), or slowed (eg. in tumours) through manipulating Sox18 activity. 20 21

Kidney development & disease

Kidneys arise via Genetic basis of kidney been shown to be critical for normal development kidney development and ongoing molecular dialogue The kidney is a complex and vital function and is mutated in a rare between two simple organ. If kidneys fail, treatment renal failure condition called Denys tissues during involves long term dialysis or organ Drash syndrome. The WT1 gene encodes many related proteins which development. These must interact to regulate both DNA create highly specialised and RNA within the cell. We are investigating how this regulatory filtration units and a protein works to create a kidney plumbing system which and keep it functioning by examining filter blood to produce what genes it regulates and what proteins and nucleic acids it interacts urine, control blood with and for to what end. pressure and red blood The Slit genes in kidney cell number. We are development examining what genes We recently isolated members of the Slit gene family from developing control this process to transplantation, both of which have kidney. These genes are related generate a normal and considerable side effects. The to a fruitfly gene which regulates projects under way in this laboratory functional kidney. The nervous system and excretory system all focus around the molecular basis development. There are three corollary of this is of kidney formation: from novel members of this gene family in understanding what gene isolation, to expression pattern, vertebrates and each has a specific protein localisation, mode of action pattern of expression during kidney goes wrong in kidney and function within the kidney. Our development suggesting roles in disease with a view aim is to ultimately impact on renal disease by first understanding to diagnosis, treatment the normal process of kidney and ultimately organ development. regeneration. Mechanism of action of the WT1 protein The WT1 gene was isolated as a tumour-suppressor gene mutated in a type of childhood kidney cancer, Wilms’ tumour. We have verified that while mutated in sporadic Wilms’ tumours, WT1 is not mutated in all such tumours. WT1 has subsequently 20 Program in Genetics & Developmental Biology 21

& disease Group leader Melissa Little development • Postdoctoral staff • Aaron Smith, Julie Scott, Gary Shooter

Students • Michael Piper, mesoderm migration and nephron therapeutic potential in kidney, lens Elida Szilagyi, Edmund Sim formation. We have established a and CNS disorders. kidney organ culture system to test Research assistants Screening for novel genes • the role of these gene products in Lorine Wilkinson, Kylie Georgas kidney . We are also involved in nephron formation End stage renal failure (ESRF) is a major health burden worldwide with 250,000 new patients in the US annually. ESRF amongst indigenous populations worldwide is also climbing dramatically for unknown reasons. An understanding of what genes are required to make a nephron provides fundamental information for the early diagnosis of those likely to suffer ESRF later in life and ultimately will enable us to regenerate functional kidneys for investigating the expression pattern the treatment of ESRF patients. and function of slit receptor proteins, To more rapidly isolate important the ‘roundabout’ proteins. genes in kidney development, we are embarking upon microarray-based high throughput expression analysis. Crim1: A novel gene involved Novel genes isolated from such in organogenesis Crim1 is another novel gene that screens can then be assessed for their we isolated from developing kidney. importance in kidney development This gene encodes a large membrane using the organ culture assays we protein with characteristic cysteine- have now established. rich motifs likely to interact with a known family of growth factors, the TGFbeta superfamily, to control, not only, kidney development but also lens, spinal cord, brain and tooth. Crim1 is also expressed strongly in the testis, but not the ovary, suggesting a role in male differentiation. We are biochemically characterising the ability of Crim1 to interact with growth factors and establishing bioassays to assess its 22 Program in Genetics & Developmental Biology 23

Molecular analysis of cutaneous systems

The skin provides a Skin and hair follicle Keratinocyte differentiation development and formation of the barrier that protects A major focus of our group is epidermal barrier us from harmful identifying the signals and messages Primary functions of the epidermis involved in specifying the formation are to protect us from environmental radiation, of the integument. We are studying assault and prevent desiccation. A environmental toxins, several genes and their protein key protein involved in both these functions is profilaggrin. This large and infection as well and abundant protein is expressed as preventing late in epidermal differentiation and moisture loss. We are is thought to regulate skin water content, have innate anti-microbial studying the activity, modulate the skin’s response contribution of to UV damage and act as a calcium sink. We have cloned the mouse individual genes and Profilaggrin gene and other related proteins to the genes. maintenance of Determination of the roles of healthy skin. An the non-helical domains of understanding of the intermediate filament proteins Intermediate filament proteins consist molecular processes of a highly conserved central involved will permit a alpha-helical domain flanked by non- helical domains of varying size and more targeted composition. While the function of approach to the the helical domain has been known treatment of inherited products including, Smoothened, and acquired skin Frizzled3 and Gli1. In collaboration diseases such as with Brandon Wainwright’s group eczema, psoriasis and we have made transgenic mice that overexpress Smoothened and Gli1 in cancer. order to determine the contribution of these genes to the development of basal cell carcinoma. We have also undertaken a systemic analysis of Frizzled gene expression during skin development in collaboration with Graham Cam (CSIRO-Prospect). 22 Program in Genetics & Developmental Biology 23

cutaneous systems Group leader • Joe Rothnagel

Postdoctoral staff • Xue- Qing Wang, Donna Mahony

Students • Lexie Friend, we have examined the of Betsy Hung, Pawel Listwan, Jonathan Beesley, Monica mouse KRT6a in transgenic mice. Kessler, Rina Massadah, Amy We found that deletions into the McCart promoter resulted in constitutive expression of the transgene but Research assistant • did not alter cell type specificity. Seetha Karunaratne We have localized the cis-elements that mediate inducibilty to region 1.5kb upstream of the transcription initiation site.

for several years, the role of the end domains has yet to be fully elucidated. We have used fluorescent tags to determine that a subset of these end domains contain motifs that specify cytoplasmic localization through interactions with microtubule filaments. This knowledge has helped explain several in vitro observations and data from knockout mice.

KRT6a: an inducible epidermal keratin The KRT6a gene encodes keratin K6a, which is normally expressed in the companion layer of the hair follicle but is induced in outer root sheath and epidermal keratinocytes in response to wounding. Because of the utility of an inducible promoter for applications such as gene therapy 24 Program in Genetics & Developmental Biology 25

Molecular genetics of pigment ation

The physical traits of MC1R alleles in normal variant most frequent in fair, blonde variation of human and light brown hair colours. These skin, hair and eye pigmentation and skin cancer three red hair variants were found to colour are important risk be overrepresented in a population- Classical genetic studies of factors determining based sample of 460 familial and pigmentation patterns between and sporadic melanoma cases compared an individuals risk within human population groups to 399 control individuals; each of of solar UV-induced have so far revealed little as to these alleles doubled melanoma risk its molecular basis, although it and increased risk fourfold if two skin cancer. We are variants were carried. investigating the Establishment of immortal human genes that are human melanocyte cell lines involved in directing of different MC1R genotype Sensitivity to sunburn and poor pigmentation tanning ability are well recognised and help risk factors for both melanoma define a person’s skin and non-melanoma skin cancer. We intend to study the functional cancer risk. The consequences of these changes on the group has isolated physiology of the melanocyte which produces the protective pigmentation and characterised induced by sun exposure. Melanocyte several human gene cell strains will be established from homologues of mouse is clear that environmental factors genes regulating coat may also play a substantial role on sun-exposed skin. Despite this colour to use them as polygenetic inheritance there is tools to investigate the likely to be only a few major genes that will exert a dominant influence molecular genetics of on pigmentation phenotype with human pigmentation. others playing modifying roles. We have studied MC1R coding region variation associations with hair and skin colour in humans by genotyping members of a large collection of adolescent caucasian twins from South-East Queensland. Twelve amino substitutions have been found at 11 different sites. Three alleles were linked to red hair with one 24 Program in Genetics & Developmental Biology 25

Molecular genetics of

pigment ation Group leader • Rick Sturm Postdoctoral staff • Neil Box

Students • Aaron Smith, Brooke Gardiner, Chris Barnes, Anthony Cook

Research assistants • Darren Smit, Wei Chen, Lisa Marks

individuals of different MC1R cell fate and gene expression. An genotype and will be immortalised antisense RNA strategy was used using the catalytic subunit of human to phenotypically knockout Brn-2 telomerase (hTERT). These cells will expression in melanoma. This loss provide a resource to investigate the of expression was associated with a role of different MC1R alleles in change in cell morphology and led to directing different gene expression the loss of a number of differentiation patterns, UV-sensitivities, and markers, including the TYR, TRP-1 establish the variables in the ability and TRP-2 pigmentation genes. The to respond to sun-exposure of cells in Brn-2 protein has been shown to culture. homodimerise in vivo with high affinity, with dimer formation The Brn-2 gene regulates the dependent on the presence of both melanocytic phenotype the homeodomain and linker regions A number of Oct-proteins that of the POU-domain by using Brn-2 interact with the control sequence deletion expression constructs. We ATGCAAAT and belong to the have also studied Brn-2 protein POU-class of transcription factors interactions with the basal have been described. The study of transcription complex, the Oct-transcription factors expressed transcriptional coactivator p300 and in melanocytic cells has identified with the Sox10 and Pax3 the Brn-2 POU domain transcription transcription factors that are known factor which may act in a similar to play an important role in role as other cell specific Oct-family melanocyte formation. members in directing melanocytic 26 27

Molecular genetics of neural devel opment

The nervous system is the most complex organ in the body Molecular genetics of neural and its formation development in vertebrates requires intricate Our laboratory is interested in the molecular genetics of neural cascades of gene development in the vertebrate regulation during embryo. During normal development, an embryo has to produce thousands embryonic and of functionally and morphologically postnatal different types of neurons and glial development. We are cells at correct positions within the nervous system and interconnected investigating the precisely with other neurons or target molecular and genetic cells. We are studying how these processes are controlled at molecular mechanisms that and genetic levels during embryonic control the development. Projects include the developing spinal cord is thought development of the control of neural cell patterning to be determined by a gradient and cell-type specification by growth of secreted signalling protein, Sonic nervous system in factors, and gene expression in hedgehog, but the exact details vertebrates. developing brain and spinal cord. of how this protein works are unknown. This project investigates Discovering how Control of neuronal cell the roles of Sonic hedgehog and its genes and molecules identity in vertebrate CNS downstream signalling pathways in by Sonic hedgehog signalling neuronal precursors in the developing function and interact pathways spinal cord. during normal The diversity of neuron types in the development will Cell-type specification by transcription factors provide a fundamental In response to differentiation signals understanding of how such as Sonic hedgehog, neuronal precursors and young neurons begin the functioning brain to express a variety of transcription is generated and factors which act as a “genetic maintained, and switch” for cell-type specification. In this project, we focus on several possible underlying transcription factors including Sox14, causes of neurological a member of the Sox family, whose expression diseases. defines unique classes of ventral interneurons in the developing spinal 26 27

Molecular genetics of neural devel opment

Group leader • Toshiya Yamada

Postdoctoral staff • Murray Hargrave

Students • Asanka Karunaratne, Gabriel Kolle, cord. To identify the exact function Annemeike Jansen, Alisa Poh of this gene, we are currently making a Sox14 gene knockout mouse model. Research assistants • Daying Wen, Ian Wilson Functional analysis of vertebrate Slit genes in developing CNS The formation of functional neuronal circuits is controlled by complex cell interactions mediated by a variety of secreted signals. We investigate this process by studying the function of vertebrate Slit homologues which are expressed in the floor plate and notochord, which are known to be critical in the control of cell

Characterization of a novel cysteine-rich transmembrane protein Crim-1 in vertebrate CNS development Crim1 is a newly discovered gene expressed in the floor plate, notochord and developing motor neuron in the spinal cord. The protein product of Crim1 is thought to regulate the functions of other growth factors in the developing CNS, in particular Bone Morphogenetic Proteins and members of the TGF- beta superfamily, through its highly conserved cysteine-rich repeats. We are studying the functions of Crim1 in the context of neuronal cell differentiation, cell migration and/or patterning and axon guidance in survival in the spinal cord. the CNS, and in developing motor neurons. 28 Program in Genetics & Developmental Biology 29

Molecular genetics of human disease

As part of our studies Function of the patched Downstream targets of tumour suppressor gene patched signalling into a common We discovered that the patched Whilst the patched forms a hetero- cancer, basal cell gene was mutated in basal cell trimeric complex on the cell surface carcinoma of the skin and in (with Smoothened and its Shh) carcinoma, we the common human brain cancer, the downstream targets of signalling isolated the gene medulloblastoma. In order to further events are largely unknown. In responsible and define the function of patched, we order to address this issue we have have modelled many of the missense established an in vitro model system showed that it was for patched/hedgehog signalling also involved in other where we can add the Shh ligand and activate the pathway. We have cancers. We are now performed microarray experiments looking at the role of with our collaborators, Drs Sean this gene and related Grimmond, Carol Wicking, and Lindsay Fowles, and have identified molecules in both several novel targets of patched/ cancer and normal hedgehog signalling which are currently under analysis to define development of the their role in cancer, differentiation embryo. For our gene and development. therapy studies we The cellular origin of basal use as our model cell carcinoma the common inherited Transgenic animal studies from several laboratories have indicated disorder cystic that patched mutant mice develop fibrosis. This work basal cell carcinoma-(BCC) like concentrates on how mutations in vitro and examined lesions. Despite their localisation and binding to the this, it is still not the gene defect results ligand, Sonic hedgehog (Shh). In clear which cells in the many collaboration with Dr Michael Fietz, of the skin give rise to BCCs. In symptoms of the some novel mutants have been over- expressed in Drosophila and as a order to address disease, and how it result we have identified a region this issue, we have might be possible to of patched which appears to be generated a responsible for binding the Shh number of correct it. ligand and the region responsible for transgenic animals transmitting the signal to the nucleus that express Cre via the G-protein coupled receptor, recombinase in Smoothened. specific skin 28 Program in Genetics & Developmental Biology 29

human disease Group leader • Brandon Wainwright

Postdoctoral staff • Elaine Costelloe, Brendan McMorran, Ian Smyth

compartments. Gene-targeted mice CF mice, and as a result it appears Students • Christelle Adolphe, Susan Gillies, Wendy have been developed which excise that the epithelial cells are the source Ingram, Christine Mulford, the patched gene in the presence of the inflammatory defect. Delvac Oceandy of Cre recombinase and current Genes controlled by CFTR breeding programs are designed to Research assistants • remove patched function from Epithelial cells lacking CFTR have a Dominic Lunn, James Palmer, specific skin compartments. dysregulated inflammatory response Emily Riley after challenge both in vitro and Origin of the cystic fibrosis in vivo. The mechanism of the inflammatory response interaction between CFTR activity Data from a number of laboratories and the inflammatory cascade is including our own has now indicated unknown, and so we have that in addition to the known approached this issue using cDNA biochemical properties of the CF microarrays. Using a 20 K human gene (CFTR), CF individuals have a cDNA chip we have profiled the CFTR-mediated inflammatory defect such that they respond massively to inflammatory stimuli. Given the cytokine profile of the infected CF lung and existing in vitro data, it appears that both epithelial and inflammatory cells may express cell autonomous CFTR-mediated defects in the inflammatory response. In order to address this question we have generated a number of transgenic mice expressing human genetic response of human airway CFTR under the control of a epithelial cells to Pseudomonas macrophage- aeruginosa in the presence and specific absence of CFTR. In this way promoter, and we have identified a number of animals genes regulated by PA and/or CFTR. expressing Some of these genes/pathways are human CFTR potential novel targets for therapeutic under the intervention into cystic fibrosis. control of an epithelium- specific element. These lines have been crossed to our 30 31

Developmental genes in human disease

The identification of genes involved in mammalian craniofacial development A common feature of many human developmental syndromes is dysmorphology of the face, the major features of which derive from the pharyngeal arches during early embryogenesis. The aim of this project is to use a large scale genomic approach to identify genes specifically expressed in pharyngeal arches and which are therefore likely to be important in the development and patterning of the face. Because facial dysmorphology is a common feature of human malformation syndromes, we anticipate that many of these genes will also play a more general role in the development of other organ systems, and will thus be implicated in a range of congenital disorders. Using the mouse as a model system, we are employing subtractive hybridisation and microarray technology to identify novel developmental genes. Access to human genome sequence information means we can more readily identify human homologues Birth defects arising from abnormal development and investigate their role in human of the embryo are a major cause of infant disease. Subcellular localisation of mortality and childhood disabilities. Our work members of the hedgehog involves the identification of novel genes involved signalling pathway The hedgehog signalling cascade in embryonic development as well as investigation plays a pivotal role in embryonic of the role of several known genes in the development and tumour formation. In collaboration with Rob Parton processes of embryogenesis and human disease. and Brandon Wainwright we have 30 Program in Genetics & Developmental Biology 31

Developmental genes

in human disease Group leader • Carol Wicking

Postdoctoral staff • Lindsay Fowles

Students • Edwina McGlinn, Tim Evans, Charlotte Webb Proteins which interact with the Gli transcription factors Research assistant • The Gli family of transcription Jenny Berkman factors are involved in mediating transcription of genes important in the hedgehog signalling pathway. There are three members of this family in vertebrates, at least two of which are involved in human disease. Using a yeast two hybrid approach, we are investigating the nature of the proteins which interact with each of these molecules. This will give us further insight into the role of the Gli proteins in development and disease.

been investigating the subcellular localisation of members of the hedgehog pathway, in particular the hedgehog receptor molecule patched. Employing both immunofluoresence analysis and electron microscopy studies, we have localised patched to endocytic vesicles within the cell. We are extending these studies to other members of the hedgehog pathway in an attempt to understand the processes involved in regulating reception of the hedgehog signal. Because cholesterol is known to play an important role in this pathway we are also investigating its possible involvement in protein localisation and trafficking. 32 33 Microarray expression profiling is a rapidly developing technology that combines robotics, molecular biology and bio-informatics to study expression of tens of thousands of genes at once in a single experiment. In this way we can identify and study genes involved in normal biological processes and those whose expression are altered in disease states.

Microarray expression profiling

Expression profiling of Redefining endothelial cell melanoma biology by expression We are using expression profiling with profiling cDNA and tissue microarrays to define Angiogenesis, or the development the genes involved in development of a vascular network, is a complex and progression of melanoma process governed by cellular tumorigenesis, define transcription interactions, changes in patterns associated with the presence of environment and signalling by key oncogenic and tumour suppressor soluble factors. Perturbation of gene mutations commonly associated this process leads to a variety with melanoma (P53, P16, PTEN, of pathologies. An ever-expanding N-RAS, B-Catenin), and analyse list of agonists/antagonists of melanoma patient samples to find angiogenesis have been defined. Group leader • Sean those genes whose expression patterns Many of these agents act directly Grimmond correlate with pathology, prognosis and on endothelial cells, key cells therapeutic response. This work is involved in generation of new Research assistant • being done in collaboration with Drs vessels. We are defining the Alistair Forrest Nick Hayward, Sandrah Pavey and transcriptional responses of David Whiteman at QIMR, and Drs endothelial cells to endogenous Jeff Trent and Mike Bittner at NHGRI, and exogenous modulators with the NIH. aim to better define the processes of migration, proliferation and Studying tumour suppressor differentiation. function of e-cadherin This project aims to identify genes Genomic and proteomic regulated by the invasion suppressor dissection of the molecular gene, E-cadherin. Cadherins are a basis of kidney development family of cell-cell adhesion molecules This work is being performed in that control tissue and cell patterning collaboration with Dr Melissa Little in most tissues of the body. In (IMB) and aims to improve our collaboration with Dr Alpha Yap understanding of mouse kidney (IMB), global gene expression development. The temporal gene profiling is being used to study the expression pattern of the action of human E-cadherin in a developing kidney and the process series of E-cadherin inducible human of mesenchyme-to-epithelial colon and breast cancer cell lines with transition is to be monitored by differing metastatic potential. expression profiling. 32 33 Cell Biology

The cell is the fundamental unit of life. Cell biologists strive to understand the complex molecular mechanisms that act together to make up complex organisms such as man. Our understanding of these processes is vital to our fight against disease. 34 Program in Cell Biology 35

Macrophages and osteo clasts

Macrophages are Control of the c-fms and TRAP genes large white blood cells Like all cell types, macrophages and that form a front line osteoclasts express a wide range defence against off lineage-specific genes that are required for specific functions. We infectious disease and have an ongoing program aimed contribute to normal at identifying the key elements of the c-fms and TRAP genes required tissue turnover in for expression in macrophages and development and osteoclasts respectively. The c-fms wound repair. gene contains a key element in the first that is highly-conserved Osteoclasts are related in mouse and human. This element cells in bone that has been shown to be in open chromatin only in macrophages. We control the release of have found that the fms intronic calcium. By regulatory element (FIRE) is both an understanding how Actions of microbial DNA required for macrophage- specific transcription and an these cells function, Mammalian macrophages are able to recognize danger signals that inducible that controls the we hope to bolster are generic to wide classes of level of expression. We have normal immunity and microorganisms, such as the cell wall succeeded in identifying the minimal component lipopolysaccharide (LPS), elements required for tissue-specific restrict their activities and respond by amplifying innate expression of fms, and created in diseases like defences and releasing regulators that arthritis, osteoporosis prime the acquired immune response. One of these “pathogen-associated and atherosclerosis molecular patterns” is bacterial DNA, where they are which contains unmethylated CG dinucleotide sequences that are responsible for much suppressed or methylated in the of the pathology. mammalian genome. We have found that replication protein A, a protein involved in DNA damage recognition and normal DNA replication, has a selective ability to bind macrophage- activating DNA sequences. The mechanisms linking this recognition to signalling in macrophages are currently being elucidated. 34 35 osteo clasts Group leader • David Hume

Postdoctoral staff • Ian Cassady, Roy Himes, Kate Stacey, Timothy Ravasi, Matthew Sweet, Kathleen Murphy, Julie Osborne transgenic mice in which the fms global analysis. We have obtained promoter drives expression of the arrays of macrophage-expressed green fluorescent protein to cells of genes and begun to analyse inducible Students • David Sester, the macrophage lineage. Studies of gene expression in macrophages. Stephen Cronau, Tedjo the TRAP gene, which encodes a key Completed studies provide insight Samono, Nicole Walsh, protein required for bone resorption, into the remarkable plasticity of Hasnawati Saleh, Stacey Wardrop, Darryl Houghton, have focussed on the regulatory gene expression in macrophages. Weng Hong Toh, Colin Cheung importance of transcription factors Subclones of the macrophage cell of the microphthalmia gene family. line RAW264 display widely We have also identified a novel divergent spectra of expression of Research assistants • TATA-element that is utilised only in macrophage-specific genes either in Elizabeth Williams, Christine osteoclasts. The study of osteoclast the basal state, or particularly when Wells differentiation has been expedited by stimulated with LPS. Clustering the identification of a subclone of a algorithms reveal families of genes macrophage cell line (RAW264) that can express osteoclast-specific genes when stimulated with osteoclast- differentiation factor.

cDNA microarrays Traditional studies of gene regulation focus on one gene at a time. The availability of large numbers of sequenced cDNAs, and technology to be array them in grids of thousands on glass slides, permits a more

that tend to be coregulated. In another study, we have used arrays to analyse the effect of a mutation of the putative receptor for LPS, TLR4. Mice with this mutation (C3H/HeJ) fail to induce a set of inflammatory genes in response to LPS, but array analysis reveals sets of genes that retain LPS responsiveness. The results suggest that either the mutation is only a partial loss of function, or there is another receptor. 36 Program in Cell Biology 37

Cell biology of diabetes

We are deciphering Insulin signal transduction A major question is how different the molecular basis growth factors achieve biological of Type II diabetes, specificity given the tremendous a disease affecting as overlap in downstream signalling pathways. The insulin receptor (IR), many as 1.5 million like many growth factors, stimulates Australians. Insulin phosphatidylinositol 3’ kinase and a serine threonine kinase (Akt) in is secreted in response muscle and fat cells as an essential to increased blood part of the metabolic program. sugar to enhance However, metabolic regulation is highly specific to IR. We are glucose uptake into currently exploring the molecular muscle and basis for this specificity by focussing Regulated exocytosis of adipocytes. This is on the intracellular location of these GLUT4 signalling events in adipocytes, GLUT4 is the major glucose mediated by a because location combined with transporter in muscle and adipocytes. complex series of strength and duration of signalling Insulin stimulates the exocytosis are essential parameters in defining of this molecule from intracellular molecular reactions specificity. In addition, we are using a vesicles called GSVs to the plasma that somehow go awry proteomics approach involving mass membrane of the cell. We have in Type II diabetes. spectrometry to discover new targets identified a targeting motif in the that are downstream of insulin- cytosolic tail of GLUT4 that controls stimulated Akt in adipocytes. early endosomal sorting. This motif is also found in a number of other proteins including an insulin responsive amino peptidase, a protein convertase and certain Syntaxins. We have recently devised a strategy for 36 Program in Cell Biology 37

diabetes Group leader • David James

Postdoctoral staff • Nia Bryant, Jon Whitehead, Sally Martin, Fiona Simpson, Juan Carlos Molero

studying the kinetic movement of and similar sets of molecules are Students • Annette Shewan, GLUT4 in adipocytes and these also found in yeast. Studies in Charlotte Widberg studies have led us to propose yeast afford enormous technical a model in which GLUT4 cycles advantages because it is possible to Research assistants • between the Golgi and the combine genetics with cell biology to Mandie Prior, Teresa Munchow, endosomes via the unique endosomal rapidly acquire detailed information. Tara Carton, John Normyle, targeting motif. Insulin somehow Using this approach we have recently Suzie Verma overrides this cycle resulting in a gained tremendous insight into the shift in the distribution of GLUT4 to mechanism by which a family of the cell surface. Future studies will proteins (Sec1 proteins) control focus on identifying the endosomal vesicle docking via the target sorting machinery that binds to this receptor protein, Syntaxin; new class of sorting motif. Docking of GLUT4 vesicles with Vesicle docking and fusion the plasma membrane in A major focus of our laboratory is adipocytes to define the mechanisms that allow We have discovered 4 molecules that discrete vesicle carriers to dock and are involved in docking/fusion of fuse with their appropriate target GLUT4 with the plasma membrane; membranes inside the cell. To do this Syntaxin4, SNAP23, VAMP2, Munc- we focus on 3 areas: 18c. Using a novel affinity purification technique we have begun to identify partners for each of these proteins as the first step to understanding their function in insulin action;

Melanogenesis We have recently discovered that Syntaxin7 is involved in late endosomal transport and is substantially upregulated in B16 melanocytes. Taking advantage of this observation we have immunopurified Syntaxin7 Vesicle transport in Saccharomyces complexes from these cells and have cerevisiae identified several new Syntaxin7 Many of the molecules that regulate binding partners including VAMP8, vesicle docking/fusion in mammalian mVti1b, alpha SNAP and a myosin cells are conserved through evolution phosphatase regulatory subunit. 38 Program in Cell Biology 39

Muscle and nuclear hormone receptors

Age and Nuclear receptor cofactor or differentiate. Ectopic expression function in skeletal studies with GRIP-1 in muscle injury-induced muscle myogenesis cells demonstrated that this SRC wasting, cachexia, Nuclear hormone receptor mediated is necessary for contractile protein muscle tumours and activation of transcription is gene expression and differentiation. associated with cofactors collectively Furthermore, we demonstrate that, hypertrophy reflect a denoted as the steroid receptor GRIP-1, coactivates myogenin- and malfunction in the coactivators (SRCs). SRCs interact MEF-2C-mediated transcription. The with p300/CBP and recruit PCAF mechanism involves direct protein- balance between to a complex that synergistically protein interactions between muscle growth and activates transcription. PCAF and MEF-2C, myogenin and GRIP-1. wasting. These p300 have also been shown to This work demonstrates that the function as critical coactivators for steroid receptor coactivator, GRIP-1, processes are adaptive the myogenic transcription factors, potentiates skeletal myogenesis. responses to exercise, MyoD, myogenin, and MEF-2 during differentiation. We examined the role Cofactor sub-cellular injury and of one SRC, GRIP-1, in muscle localisation in muscle and pathological stimuli. differentiation, an ideal paradigm rhabdomyosarcoma cells for the analysis of the events that GRIP-1 is necessary for skeletal Our group focuses on govern the cell’s decision to divide myogenesis, and functions as a investigating the role cofactor for the transcription factor MEF2. We localised GRIP-1 in of nuclear hormone the nucleus of proliferating receptors and myoblasts, however, notable associated cofactors in expression was observed in the cytoplasm. Differentiation induces a muscle development as predominant localisation of GRIP-1 a critical step toward to the nucleus. MEF2 is primarily expressed in the nucleus, although elucidating the we observed a mosaic or variegated function of these expression pattern in myoblasts. proteins in muscle However, all nuclei in myotubes express MEF2. Rhabdomyosarcoma disease. (RMS) cells derived from malignant skeletal muscle tumors express GRIP-1 in a diffuse nucleo- cytoplasmic staining pattern. MEF2 and the other SRC cofactors, SRC-1 and SRC-3 are abundantly expressed in RMS cells, however, the staining is not localised to the nucleus, and 38 Program in Cell Biology 39

nuclear hormone receptors Group leader • George Muscat

Postdoctoral staff • Uwe Dressel, Jonathan Harris

Students • Mary Wang, demonstrated that mutations in the Senali Abayratna-Wansa, Brett Hosking, Tom Chen, Paul Sry-related HMG box gene Sox18 Rohde underlie cardiovascular and hair follicle defects in the mouse mutant Research assistants • ragged (Ra) and the allelic mutant Shayama Wijedsa, Udani RaJ. MEF2C is expressed in Abeypala developing endothelial cells. Deletion of the Mef2c gene in mice results in vascular abnormalities with some similarity to the Ra defects. We show direct interaction between the MEF2C and SOX18 proteins which results in synergistic trans-activation. This interaction is mediated by the MADS domain of MEF2C, and by two regions of the SOX18 protein, the HMG domain, and a carboxy terminal region that includes the activation domain. In situ hybridization and immuno- fluorescence established that MEF2C and SOX18 are expressed and colocalised in endothelial cell nuclei. The Ra and the RaJ mutant SOX18 proteins fail to interact with MEF2C suggesting that this physical predominantly accumulates in the association has important cytoplasm. This suggests trafficking consequences for vascular of transcription factors and steroid development. receptor coactivators is a critical event during myogenesis. More importantly, the breakdown in SRC trafficking in rhabdomyosarcoma cells suggests this process has a critical function in differentiation and the onset of tumorigenesis.

Sox18, and the regulation of endothelial gene expression In collaboration with Peter Koopman’s laboratory we 40 Program in Cell Biology 41

Cell biology of the plasma membrane

Animal cells are Structure and function of caveolin identified with the SV40 caveolae screen were tested for their effect enclosed by a Caveolae are a characteristic and on specific signalling pathways (in membrane, termed abundant feature of the plasma collaboration with Prof. John the plasma membrane of many mammalian cells. Hancock). A caveolin mutant These 60-80nm uncoated flask- specifically inhibited signalling by membrane. This shaped pits have been implicated in one Ras isoform, H-ras, but not by membrane acts as a many different cellular processes but a different isoform of Ras, K-ras. their exact function remains unclear. Unexpectedly, the mutant caveolin barrier to stop The major membrane proteins of protein was shown to be acting invasion by caveolae are integral membrane through an effect on cholesterol pathogens while proteins termed caveolins. Caveolins transport and, as a consequence, have been linked to a number inhibited signalling events occurring allowing of disease conditions, including in cholesterol-rich cell surface communication tumorigenesis, Alzheimer’s disease, domains. This study provided the first Niemann-Pick type C, and limb- evidence that H-ras signalling occurs between the outside girdle muscular dystrophy. A major in specialised domains of the cell environment and the focus of our group is to understand surface termed ‘lipid-raft’ domains. interior of the cell. the role of caveolae and caveolins in We are currently studying how cellular function. caveolin regulates lipid transport, an We are studying how area of intense interest at the present the plasma membrane Entry of viruses into animal time. cells functions in healthy We have shown that the tumour virus, Muscle development and cells, how toxic SV40, hijacks caveolae as a means to muscular dystrophy gain entry into the cell and ultimately Muscular dystrophy is characterised agents such as viruses to reach the cell nucleus where it by increased membrane fragility and have managed to replicates. We are now characterising disorganisation of surface domains in overcome the cell’s the trafficking pathway. We have also muscle cells. We discovered a major examined the role of caveolins in component of muscle caveolae, now defences, and how the SV40 entry by generating mutant called caveolin-3. Two recent studies normal functions of caveolin proteins and assaying their suggested that specific mutations in effects on the infectious entry caveolin-3 occur in patients with the plasma membrane pathway. In this way we were a type of limb girdle muscular are disrupted in able to generate dominant negative dystrophy. In mature muscle diseases such as inhibitory mutants of caveolin which caveolin-3 is an abundant protein inhibited viral infection. of the muscle plasma membrane muscular dystrophy where it may play a role in the and cancer. Functional studies of functional organization of the muscle caveolins fibre surface. We analysed the effects Dominant inhibitory mutants of of dystrophy-associated mutations 40 Program in Cell Biology 41

plasma membrane Group leader • Rob Parton Postdoctoral staff • Amanda Carozzi, Albert Pol, Johanna Gustavsson, Sharon Clark, Pierre-Francois Mery

on the localisation of the caveolin developed. We are using a number Students • Susan Nixon, Ayanthi Richards, Isabel protein and on caveolar function. of novel tools for examination of Morrow, Cedric Asensio Caveolin-3 trafficking is blocked in lipid distribution at the electron cells expressing one of the mutant microscopic level. Combined with Research assistants • proteins. Using the Ras signalling on-section labelling techniques, these Robert Luetterforst, Charles assay we also showed that a second agents are giving the first insights Ferguson, Adrian Knight, mutant might perturb lipid raft into the distribution of Kathryn Green domains and so disrupt specific lysobisphosphatidic acid, an antigen signalling pathways. associated with an autoimmune

Electron microscopic disease, phosphatidylinositol-3- localisation of lipids phosphate, which regulates Specialised domains of lipids, such membrane traffic, and as lipid rafts, have been shown to sphingomyelin, a marker of lipid raft be of crucial importance for many domains. important physiological processes. However, the tools and techniques for lipid localisation are not well 42 Program in Cell Biology 43

Protein trafficking

All cells of the body Regulation of protein gene expression and imaging of trafficking live cells we are following protein have complex A major focus of our work is transport in ‘real time’, and further trafficking pathways to define the functions of some characterising the transport carriers for moving and of the genes that regulate protein and the roles of specific vesicle- trafficking in cells. This work associated proteins. Our goal is to regulating proteins. involves determining the location We are studying and function of selected proteins using a combination of biochemical protein trafficking in and microscopy techniques. We have epithelial cells, in now mapped extensive sets of signal transduction, cytoskeletal, structural organs such as the and lipid modifying proteins found kidney and gut, and on different transport. In particular in macrophages. Our we have shown how G proteins and regulators of G protein signalling studies aim to identify (RGS) proteins associate with molecules that specific membrane domains and transport vesicles, providing insights regulate normal into how these protein families might trafficking and can interact. Several new RGS proteins develop a molecular map of the secretory pathway and determine be developed as drug that have the potential to link cell growth and trafficking are also being how movement of proteins to the cell targets for the studied. With Paul Gleeson (Monash surface is regulated. University) we have defined the treatment of cancer Cadherins and cancer specific membrane domain of a Our work in epithelial cells led us and inflammation. Golgi localisation domain that targets to study how vital cell adhesion proteins to specific vesicles. Using proteins- cadherins- are trafficked. 42 Program in Cell Biology 43

Group leader • Jennifer Stow

Postdoctoral staff • Fiona Wylie, Wenda Shurety, Lubomira Jamriska

Students • Chris Le, John E-cadherin has important roles during Cytokine secretion in Lock, Shannon Joseph, Kevin macrophages early development and in the Miranda function of many organs. The loss Secretion of cytokines such as tumour necrosis factor (TNF) is a of E-cadherin is an early event in Research assistants • metastasis of breast, colon, prostate critical part of the host defence Darren Brown, Tatiana and many other cancers indicating it against bacterial invasion and the Khromykh, Juliana Venturato is also a potent tumour suppressor. growth of tumour cells. Our In work with Alpha Yap (IMB) we group is characterising the pathway have identified a new pathway and molecules that regulate TNF for endocytosis of E-cadherin and secretion in macrophages. These have shown how its uptake and studies first described the pathways function can be regulated by for exocytic and endocytic trafficking signalling of cleaved, pathways. We biological are also active TNF, studying and we E-cadherin have now trafficking also and described expression in the breast cancer disparate cells with the trafficking ultimate aim of of developing uncleaved new forms of manipulations TNF. This or strategies latter study for tumour has suppression. provided More recently new with Rohan insights into Teasdale current (IMB) we have begun to characterise clinical strategies focussed on using targeting signals in cadherins, using proteinase inhibitors for controlling bioinformatic and mutagenesis excess TNF. In current work we approaches, with the first are studying gene expression in the identification of a signal in cadherins secretory pathway of macrophages for polarised trafficking to the cell with the aim of finding strategic, surface. pathway-specific regulators. 44 45

Molecular physiology of growth hormone

Growth hormone and Receptor signalling constitutively active receptor by mechanisms substituting the external domain with related cytokine Since cloning the growth hormone a fos-jun zipper. The latter two hormones regulate (GH) receptor collaboratively, we findings have potential in livestock have have studied the mechanics production. many important of its signalling extensively through physiological Genetic programs processes such as regulated by GH postnatal growth, Our group was the first to publish on the use of metabolism, gene arrays in GH action, reproduction, identifying a number of important hepatic targets of lactation and immune acute GH action. Using function. We study the suppression subtractive molecular basis for hybridisation we have studied gene expression this regulation at all during the earliest phase levels, from the cell of differentiation from fibroblasts to adipocytes as surface receptors to initiated by growth. This the genetic programs has identified several GH used. Research targets intimately involved in differentiation, and outcomes have supports our view that implications in the GH is involved in this process during fields of livestock embryogenesis. Aspects of production, clinical mutagenesis of hormone and this are being studied with treatments for short receptor, through the use of the use and creation of specific GH monoclonal antibodies specific to receptor knockout mouse models. stature and giantism, the receptor, and through model diabetic kidney peptide systems. We have evidence Direct nuclear actions of GH disease and blindness, that a specific conformational change receptor in the extracellular domain of the We have shown that the extracellular periodontal disease, receptor can differentially affect domain of the GH receptor can be and for several cellular signalling pathways. We have nuclear localized, and that targeting characterised the epitope and binding the receptor to the nucleus can aspects of ageing. characteristics of the monoclonal sensitize the cell to GH action. antibody to receptor able to stimulate We have shown the GH receptor growth. We have also created a 44 Program in Cell Biology 45

growth hormone Group leader • Mike Waters Postdoctoral staff • Richard Brown, Patrick Lau

Students • Becky Conway- Campbell, Jenny Rowland, Yvonne Sau Ping, Cathy Shang, extracellular domain can act as If induced by other factors, they Yu Wan a gene transactivator, and have can result in tissue resistance to identified putative interacting nuclear the anabolic actions of cytokine Research assistants • proteins using proteomics hormones, such as occurs in Dion Auriac, Kirstin Millard, approaches. We believe that nuclear inflammation. We have evidence Thea Monks, Lida Stjepkovic, GH receptor (extracellular domain, SOCS genes are used to shut down Mary White cleaved from the receptor) serves to lactation when the gland fills, and reinforce the signal coming from the that they may control hypothalamic cell surface receptor in response to feedback sensitivity to prolactin. We GH binding. also find expression of SOCS genes are elevated in breast cancer, and Role of suppressors of one SOCS is constitutively activated Cytokine Signalling (SOCS) in in all breast cancer lines examined. breast cancer and lactation This may be important in preventing SOCS genes are rapidly induced immune cytokine-mediated attack on by GH and the closely related breast cancer cells. hormone of lactation, prolactin. They induce resistance to further hormone stimulation in a feedback mechanism. 46 Program in Cell Biology 47

Cell adhesion and morpho genesis

The organization of Coordination of cadherin with more stable actin bundles. This activity and the actin implies that there is dynamic interplay different tissues in the cytoskeleton between cadherin adhesive binding body depends upon It is becoming increasingly clear and the organization of the actin the ability of cells to that early adhesive recognition cytoskeleton, which is likely to be between cells entails the close critical to cell patterning and recognise and adhere coordination between cadherin recognition. We are now studying to one another. Our activity and the actin cytoskeleton. the molecular mechanisms which We have developed assays that may mediate the interplay between research aims to allow us to analyse the relationship E-cadherin and actin dynamics. understand how between cadherin presentation and the organization of the actin Role of Rho family GTPases in adhesion molecules cytoskeleton. We have found that cadherin function mediate cellular E-cadherin associates with two A major goal of our work is to recognition and tissue quite distinct states of the actin understand how the morphogenetic cytoskeleton. In the earliest effects of E-cadherin are controlled at patterning. We focus contacts, E-cadherin co-localizes the cellular level. We have identified on E-cadherin, which with actin filaments that are actively a critical role for the Rac GTPase is essential for polymerizing, but later associates in controlling early cadherin adhesive epithelial organization and acts to retard tumour progression. Elucidating how E-cadherin controls cell recognition and morphogenetic movements will provide a basis for understanding tissue patterning in health and disease. 46 Program in Cell Biology 47

morpho genesis Group leader • Alpha Yap Postdoctoral staff • Eva Kovacs

Students • Andrew Paterson, Radiya Ali, Ailsa McCormack

Research assistants • Marita Goodwin, Lucia Ha

events. Rac activity is essential to proteins such as cadherins, play a coordinate cadherin presentation and critical but ill-understood role in actin polymerization necessary for their biological activity. We recently cells to make their first adhesive demonstrated that the lipid kinase contacts. Furthermore, the formation PI3-kinase, is necessary for cadherin of cadherin-based contacts may mark activity, implicating membrane lipids the sites for rac signaling to occur. in cadherin function for the first time. We are currently testing the notion The transient association between that E-cadherin determines the site PI3-kinase and E-cadherin may be for activation of downstream signals essential for cells to make contacts that link Rac activity to the actin with one another, marking sites cytoskeleton. for stabilization of adhesion and reorganization of the actin Role of the membrane lipid cytoskeleton. environment in cadherin regulation The lipids that surround membrane 48 49 48 49 Structural Biology

We are determining the three-dimensional structures of proteins implicated in a variety of physiological functions and disease states. An understanding of protein architecture provides the foundations for a range of drug design programs. 50 Program in Structural Biology 51

NMR in drug design

The function of a NMR as a tool in drug design to overcome current limitations on protein depends on A major focus of our group is the the use of conventional proteins as use of small disulphide-rich proteins drugs, i.e., their poor bioavailability its three-dimensional as leads in drug design. Such proteins and susceptibility to degradation in shape. We use often have potent biological activities vivo. In addition to core programs and, because of their cross-linking in protein structure determination Nuclear Magnetic disulphide bonds, usually have well we use NMR to study protein Resonance (NMR) folding, protein spectroscopy to dynamics, and ligand binding interactions. determine the structures of key Anti-microbial peptides and proteins implicated in proteins disease states. There is an urgent need for the Knowledge of protein development of new structures allows us antibiotics, given the widespread to understand their development of function and to microbial resistance to design drugs that may existing drugs. We are exploring the potential moderate that of peptide-based function to block molecules as therapeutics. To this disease. Reports from end, we have recently native medicine determined the three- applications often defined three-dimensional structures dimensional structures of MiAMP1, that can be determined using NMR an antifungal protein from provide us with leads spectroscopy. The proteins we study macadamia nuts, RK1, an for interesting come from animal and plant sources, antibacterial molecule from rabbit as well as “designer” proteins we kidney, and drosocin, an antibacterial proteins from plants produce in the lab. In particular peptide from the haemolymph of and animals, and we we have been exploring the fruit flies. These studies involve mix our lab work with bioengineering of circular proteins. collaborations within the university By cyclising proteins and creating (MiAMP1, CRC for Tropical Plant field trips to discover embedded knots within the structures Protection), nationally (RK1, Howard new natural products. using disulphide bonds, we are Florey Institute, Melbourne) and able to significantly enhance the internationally (Wistar Institute, stability of proteins. Our goal is USA). 50 Program in Structural Biology 51

drug design Group leader • David Craik Postdoctoral staff • Richard Clark, Norelle Daly, Martin Scanlon, Horst Schirra

Students • Daniel Barry, Julie Dutton, Michael Circular proteins, knots and a consensus three-dimensional fold, Korsinczky, Ailsa McManus, natural products called the cyclic cystine knot. The Jason Mulvenna, Mariel We recently discovered and complex topology of these molecules Quimio, Johan Rosengren, characterized an exciting new family also allows us to explore Manuela Trabi, Clement Waine of plant proteins that have the mathematical knot theory within the unusual features of a cyclic backbone framework of natural proteins. Research assistant • and a knotted arrangement of Shane Simonsen disulphide bonds. A major program Structures of protein-based based on this discovery involves venom components and plant Support staff • Karl Byriel, fieldwork in Cape York, protein toxins Cameron Jennings, Fung Lay, Peter Nilsson, Johan Svensson isolation and sequencing, protein Many animal venoms and plant structure determination and protein toxins contain disulphide-rich bioengineering. We have named this proteins that are valuable leads in novel family of proteins the plant drug design. Our group uses NMR cyclotides and are exploiting the to determine the structures of these exceptional stability of these molecules, with recent examples molecules as frameworks for the including conotoxins from marine delivery of peptide based cone snails, spider toxins from therapeutics. We have now the Australian funnel web spider determined the structures of several and Mexican red-knee tarantula, a family members and have determined protease inhibitor from the skin secretions of the frog Bombina bombina, and a newly-discovered protein from the venom of Australian taipan snakes. Our goal is to understand structure-function relationships of these toxin molecules to design new drug leads. In related studies in collaboration with Dr Marilyn Anderson of La Trobe University, we are determining the structures of a series of plant-derived protease inhibitors that have potential applications as insecticidal agents. 52 Program in Structural Biology 53

Structural basis of protein interactions

The formation of Intrasteric (active The available data suggest that site-directed) regulation intrasteric regulation is emerging interactions between Regulation of protein function is as an increasingly important macromolecules is a vital for the control of cellular regulatory mechanism in most crucial part signal processes. Allosteric regulation is cellular processes. a well established mechanism of transduction, protein regulation, describing Phenylalanine hydroxylase apoptosis, effectors that bind to regulatory Phenylalanine hydroxylase (PAH) is sites distinct from the active sites a metabolic enzyme, the defects transcription, cell-to- and alter protein function. We have in which cause the disease cell communication been studying a number of proteins phenylketonuria, one of the first and protein folding. including protein kinases, metabolic characterized genetic diseases that enzymes (phenylalanine can lead to mental retardation and We are interested in hydroxylase) and receptors (nuclear death. PAH is therefore tightly understanding the transport factor importin-alpha) regulated in vivo. We used X-ray which are instead regulated through crystallography, nuclear magnetic mechanisms intrasteric, active-site directed resonance and mutagenesis to underlying such mechanisms, representing the understand the structural basis of its interactions, through counterpart of allosteric control. complex regulation, which we show includes intrasteric control. employing a combination of structural, biophysical and molecular biology techniques. Our studies will lead to a detailed understanding of cellular processes, and uncover new ways to design therapeutic agents that manipulate these processes in disease. 52 Program in Structural Biology 53

protein interactions Group leader • Bostjan Kobe Postdoctoral staff • Helen Blanchard, Dianne Keough, Yingmei Qi, Ross Brinkworth

Student • Eva Golob Importin-alpha and nuclear mechanism, whereby a signal is import transmitted from the receptor binding Research assistant • Trazel Teh In eukaryotic cells, proteins need subunit to induce the fusion-activated to be imported into the nucleus. conformation of the transmembrane Such transport is directed by special subunit. The structure of the protein signals termed the nuclear is now being exploited to develop localization sequences (NLSs). new therapeutics using structure- Importin-alpha is the nuclear import based drug design. receptor that recognizes these NLSs. To obtain a complete overview of Solenoid proteins interactions formed by importin- Structural biology hopes to uncover alpha during the essential process the relationships between amino acid of nuclear import, we determined sequences, three-dimensional (3D) the three-dimensional structures of structures and protein functions. importin-alpha and its complexes Solenoid proteins, containing a with NLSs, and kinetically and superhelical arrangement of repeating thermodynamically characterized the structural units, convey the least interactions. The process of nuclear complicated relationship between a import is shown to be regulated sequence and the corresponding 3D through an intrasteric mechanism, structure. These proteins have and coordinated through an intricate particularly useful characteristics for set of interactions with varying forming protein-protein interactions, affinities. as exemplified by the nuclear import factor Retroviral envelope proteins importin- Enveloped viruses fuse their alpha, a membranes with host cell membranes protein to enable the transfer of their genome built from into the host cell. To understand armadillo the molecular basis of retroviral repeats. membrane fusion and explore novel The strategies for therapeutic design, we repetitive studied the envelope protein from structure also HTLV-1 (human T cell leukemia facilitates 3D structure prediction, virus type 1), the virus associated which can illuminate protein with adult T-cell leukemia and function, as we show in the example tropical spastic paraparesis. The of Leishmania surface protein combination of crystal structure containing leucine-rich repeats, analysis and site-directed proteophosphoglycan. mutagenesis revealed the fusion 54 Program in Structural Biology 55

Protein structure and drug design

The ability to see Bacterial disulphide proteins A long standing aspect of our what a protein looks research is the study of bacterial like at atomic disulphide catalysts that help to fold resolution allows us secreted proteins. Many of these Dsb proteins have been found to to understand how be important in the production of a protein functions bacterial toxins and virulence factors, and so may represent therapeutic in the normal state, targets for antivirulence drugs. In and malfunctions in collaboration with Dr Linda Thony- a disease state. Most Meyer (ETH Switzerland) we have determined the structure of DsbE, a importantly, each protein disulphide reductant, at 1.2 newly derived Å resolution. This will be used in a structural comparison of proteins structure of a protein that incorporate a thioredoxin fold represents a quantum to investigate protein evolution and leap in our ability to form the basis of structure-based antivirulence drug design. to develop novel drugs by structure- Adrenaline synthesis based design In collaboration with Professor Gary Grunewald (University of Kansas) are currently using this structure to technologies. Current and Dr Michael McLeish (University design potent and selective inhibitors. research centres on of Michigan) we have determined the crystal structure of the adrenaline Sulphonating enzymes the areas of diabetes, synthesising enzyme, PNMT, and In collaboration with Mick McManus hypertension, drug (Physiology UQ) we are metabolism and investigating the structures of several human antimicrobials. sulfotransferase enzymes that metabolise xenobiotics. Two of these proteins have been purified to homogeneity and initial crystallisation trials have yielded promising results. 54 55

drug design Group leader Jennifer • Martin

Postdoctoral staff • Tracy Arakaki, Fiona McMillan, Scott Rowlinson, Ulrika Rova

In collaboration with Paul Young SNARE proteins involved in Students • Melissa Edeling, (Microbiology UQ) and David Fairlie glucose transport Alan Lowe (U Bath), Francois (IMB) we are investigating the In collaboration with David James Terblanche (U Pretoria) structure of the Dengue virus and Judy Halliday (IMB) we are protease, a target for the design studying SNARE proteins that Research assistants • regulate the trafficking of the of antivirals. During this year we Cath Latham, Christine Gee have successfully overexpressed and insulin-regulated GLUT4 glucose purified the protease and developed transporter to the plasma membranes of adipocytes. We have produced Scientific support • Karl in vitro assays to identify suitable Byriel inhibitors. Crystallisation studies are two of the proteins involved in the currently underway.

HIV Virus Over the past few years we have determined the structures of several complexes of HIV-1 protease with designed inhibitors. This year we have determined the high resolution structures of substrate-bound and product-bound forms of the HIV-1 protease enzyme. The work is a collaboration with David Fairlie and Paul Alewood (IMB), and the regulation of GLUT4 trafficking, structures were determined by Len in quantities sufficient for Pattenden, a PhD student in David crystallisation trials, and we have Fairlie’s group. The begun to investigate their three-dimensional information could intermolecular association using yield vital information on how to biosensor technology. combat the HIV virus, particularly with respect to viral resistance. 56 57 56 57 Biological Chemistry

Metabolic functions essential for life in a complex organism involve a multitude of chemical reactions. We exploit the diverse chemistries of living systems in the pursuit of a better understanding of how the organism works. This understanding will allow us to design and produce new and better drugs. 58 Program in Biological Chemistry 59

Bioactive peptides and proteins

A major focus in my laboratory is the use of synthetic chemistry to determine the roles that proteins and peptides play in living cells. Solid phase Discovery, synthesis and protein engineering, and in suitable structure-function cases may be used as templates for technologies and relationships of Australian drug design. Current projects focus chemistry are continually toxins on toxins which target ion channels, Small disulphide-rich proteins have G-proteins and transporters. being developed to give a variety of diverse functions in ready access to milligram both plants and animals, but are Chemical synthesis of quantities of protein commonly found in animal venoms. proteins These venoms are often used in Large biotechnology-based domains, bioactive prey capture or defense and comprise initiatives, like the human genome peptides and the vast majority of toxins from project and other major sequencing Conus species, snakes, ants, stone- projects, are rapidly providing an structurally-constrained fish, platypus, scorpions and spiders. increasing array of new protein toxins. Such molecules These small proteins range from sequences. An outcome will be typically interact with approximately 10-120 amino acids an explosion in the number of and typically contain 2-8 disulphide gene-encoded proteins that are receptors, ion channels bonds, which play a key role in considered novel or important drug and enzymes and provide stabilising and shaping the proteins. or agricultural targets. We are They bind to their receptors with addressing one of the first tools, reagents and high affinity and specificity, and requirements for the study of these possibly drug leads for the binding residues are situated on new targets, namely the reliable our research programs well-defined surface loops and other and rapid access to the proteins of secondary structures. Because they interest in micro- to multi-milligram in cancer, pain and have such well defined structures quantities. The total chemical inflammation. they are excellent candidates for synthesis of proteins provides a rapid 58 59

Bioactive peptides and proteins Group leader • Paul Alewood

Postdoctoral staff • Peter Cassidy, Paramjit Bansal

Students • Chris Armishaw, and effective route for the production phase analogue synthesis. Solid David Wilson, Bryan Fry, Doug Horton, Lita Imperial of homogenous proteins free of supported chemistry (where starting biological contaminants, while at the materials are reversibly bound to same time offering flexibility through an insoluble polymer), with its the incorporation of unnatural amino distinctive operating principles, can acids or other chemical modifications achieve such uniformity of synthesis that improve protein efficacy. and work up procedures. Furthermore, chemical synthesis allows for facile introduction of The proficiency of solid phase biochemical and biophysical probes chemistry in library synthesis is well that cannot be made by biological demonstrated in peptide chemistry methods. Analysis of sequence data where millions of oligopeptides from the genome projects indicate (peptide libraries) can be assembled that the average protein protein is in a few weeks. We have recently approximately 300 residues, while developed methods that apply novel functional folding domains usually combi-chemistry to a range of consist of about 150 amino acid bioactive toxins and small molecule residues making them a viable target alkaloid-like targets. for chemists.

Solid phase combinatorial chemistry Chemical drug lead discovery and optimisation have traditionally been achieved in a linear fashion using solution phase chemistry, where analogues are synthesised one by one, and then screened in a relevant bioassay. Different analogues generally have wide-ranging physical properties due to the nature of the different substituents. Consequently, synthetic procedures often require modification for the successful synthesis of each analogue (or corresponding intermediate), especially with regard to the work up procedures and solvents used in the synthesis. This lack of uniformity between analogues usually prohibits simultaneous and rapid solution 60 Program in Biological Chemistry 61

Chemistry and human thera peutics

Chemistry underpins Chemical synthesis, structure investigations of chemical and and mechanism biological reactions, enzyme kinetics, every facet of the Our principal thrust is the design cell biology, and molecular molecular biosciences. of new molecules, the development pharmacology in vitro and in vivo. of new methods to make them, the Understanding how determination and analysis of their Protein surface mimics molecules interact, how structures, mechanistic investigations A key strategic objective has been structure influences of their reactivities, the use of to mimic small bioactive protein small molecules to probe biological reactivity, and how surfaces that are recognized by chemical reactions work, enables design and synthesis of reactive and bioactive compounds, and a better understanding of the molecular basis for biological processes, processes, and to elucidate other proteins, DNA or RNA during disease development, relationships between chemical disease processes. We have identified and drug action. We structure and disease regulation. In peptide conformations that are 2000 we used a range of solution common recognition elements for invent new molecules and solid phase organic synthesis protease enzymes, for classes of G and develop them into methodologies to synthesise protein-coupled receptors, and for potent enzyme bioactive molecules (e.g. de novo transcriptional designed drugs, natural product receptors. One inhibitors and receptor analogues, and peptidomimetics), of our antagonists, as reactive intermediates, highly synthetic functionalised molecular templates, chemistry potential drugs for structural constraints, and artificial programs has cancer, viral and receptors. We also synthesised metal been aimed at parasitic infections, complexes as folding templates, as developing catalysts for organic reactions, and generic inflammation, and as models of metalloproteins. We technologies Alzheimer’s disease. are engaged in design and structural for creating analysis, determination of chemical small organic structures by NMR spectroscopy and molecules to X-ray crystallography, spectroscopic mimic 60 Program in Biological Chemistry 61

human thera peutics Group leader • David Fairlie Postdoctoral staff • John Abbenante, Chris Clark, Matthew Glenn, Karl Hansford, Darren March, Robert Reid, Yogendra Singh, Martin Stoermer

Students • Rodney Cusack, Huy Hoang, Michael Kelso, Donmienne Leung, Len Pattenden, Lavinia Proctor, Robert Sbaglia, Joel Tyndall

elements of protein structure, such as and selective antagonists and agonists beta-strands, beta and gamma turns, of G protein-coupled receptors that beta-sheets, alpha-helices, helix protrude from human cellular bundles, and multi-loop bundles. This membranes and mediate cell effort has resulted in numerous small signalling. Multiple classes of small organic molecules that we have orally active organic molecules demonstrated to be both structural developed in our labs have been and functional mimics of bioactive shown to possess antitumour, protein surfaces. antiparasitic (against malaria, giardia, and schistosomal proteases), Drug design and development antiinflammatory, antiviral (low The generic resistance inhibitors of HIV and technologies Dengue proteins), and anti- that we have Alzheimer’s activities. These developed to compounds are all in various stages date have of pharmacological development in already led us animal models of human diseases. to successfully design potent and selective inhibitors of aspartic, serine, metallo and cysteine proteases, as well as potent 62 Program in Biological Chemistry 63

Glycobiology

We are studying the Development of a technology parts of the naturally occurring metabolic pathways platform for the production of biological synthesis machinery. This sialic acid-containing technology will open up necessary for the oligosaccharides opportunities for the cost effective addition of We have an established collaboration production of carbohydrate-based with Alchemia Pty Ltd, a Brisbane molecules. A successful outcome carbohydrates to based biotechnology company whose from this R&D project will have cellular proteins and core business is the chemical many long term benefits including synthesis of oligosaccharide- (or lipids. These the development of new drugs and carbohydrate-) based molecules have other products, which will in turn glycosylation events not been fully exploited as potential help to build the biotechnology drugs, mainly because they are industry in Australia. are important for a difficult and expensive to produce number of cellular using synthetic methods and to Structural studies on SNARE functions including: extract from their natural sources. proteins involved in insulin The collaborative project between action cell to cell the IMB and Alchemia is aimed This project aims to understand the communication; at developing technology that will structure and function relationships combine chemical synthesis with between the SNARE proteins that are binding of ligands to involved in the receptors; normal cell transportation growth and of the GLUT4 molecule to the differentiation; cell surface in interactions between response to stimulation by cells and invading insulin. This pathogens. Unusual pathway is glycosylation patterns important in maintaining in cells are associated correct blood with inflammation glucose levels. Novel and diseases such as treatments for cancer. diabetes could be developed from this work. [Collaborative project with Prof. David 62 Program in Biological Chemistry 63

Group leader • Judy Halliday

Postdoctoral staff • Alison Franks

Students • Ylva Strandberg, Nicholas Drinnan, Trudy Bond, Katharina Stummeyer

Technical assistants • Andrew Pearson, David Ireland, Rodney Martin

James, (IMB), Dr Jennifer Martin, Marine glues IMB, Dr Ulrika Rova, (IMB) and Dr These studies are focused on the Scott Rowlinson, (IMB)] specialised glue proteins that some marine parasites use to attach to Novel proteases involved in their hosts. These specialised proteins natural product biosynthesis have many interesting This project involves the physicochemical properties and could characterisation of the specific potentially form the basis for the proteases present in the venom ducts design of specialised glues for use in of cone snails that are important industry and medicine [Collaborative for the production of the bioactive project with Dr Ian Whittington molecules found in the venom. (Microbiology and Parasitology) and [Collaborative project with Trudy Dr Bronwen Cribb (Centre for Bond (IMB), Prof Paul Alewood Microscopy and Microanalysis), (IMB), Dr Jenny Martin (IMB) and Tamarind Hanwood (IMB and Dr Terry Walsh (QUT)]. Microbiology and Parasitology)]. 64 65

Venom peptides to drugs

Mini-proteins in the Neuronal inhibitors for the have found that CVID has a widened venoms of Australian control of pain therapeutic window between side The supply of pharmaceuticals for effects and antinociception. cone snails pain relief is a multi-billion dollar Importantly, this difference does not (conopeptides) are a industry. The narrow therapeutic arise from greater N-type vs P/Q- basis of action of existing drugs type selectivity, but likely stems from rich source of new means that the treatment of many differences in the ability of these molecular probes. forms of chronic pain is poor. omega-conotoxins to inhibit different Our group isolates We recognise this situation as an N-type VSCC splice variants. The excellent opportunity to develop new broad goals of our research are conopeptides with treatments for pain management. The to develop improved interesting features, end product from the research will be omega-conotoxins for better pain and a conopetide that selectively inhibits stroke management based on an which are then sensory nerves and can be used understanding of the precise site(s) synthesised and their for the treatment of chronic and and mode(s) of action of CVID. precise mode of neuropathic pain. Such a drug could be given to patients who no longer, or Alpha-conotoxin probes of the action and structure have failed to, respond to morphine nicotinic acetylcholine and other painkillers. receptor determined. Some of New alpha-conotoxins have been the conopeptides we Omega-conotoxin probes of discovered in the venom of have discovered have N-type calcium channel Australian cone shells which have the subtypes unique potential to distinguish among the necessay Voltage-sensitive calcium channels the nicotinic acetylcholine receptor characterisitics to be (VSCCs) play an important role (nAChR) subtypes expressed in in regulating the excitability of neurones. We are now determining developed as drugs neurones and in triggering the potency and selectivity of native or drug leads. Some neurotransmitter release. The N-type and mutant alpha-conotoxins using of these leads have VSCC has emerged as an exciting voltage clamp techniques in Xenopus new target for pain following the oocytes expressing different neuronal been used to establish successful treatment of morphine- nAChR subunit combinations. Alpha- the spin-out resistant and post-operative pain with conotoxins that differentiate among omega-conotoxin MVIIA these nAChR subunits will be used Biotechnology (Ziconitide). We have recently to characterise the native nAChRs company Xenome. discovered a new series of potent expressed in parasympathetic omega-conotoxins from Australian neurones using whole cell patch cone snails. One member of this clamp techniques. This study series, CVID (AM336), has been provides a basis for understanding identified as the most N-type vs the structural features which confer P/Q-type selective peptide known alpha-conotoxin selectivity. (provisional patent PP8419/99). We 64 65 to drugs Group leader • Richard Lewis

Postdoctoral staff • Jorgan Mould, Annette Nicke, Denise Adams

Students • Iain Sharpe, Brett Hamilton, Tina Schroeder, Demitra Temelcos, Takahiro Yasuda, Lotten Ragnarsson, Vicky Wang

Research assistants • Marion Loughnan, Trudy Bond, Linda Thomas, Caroline Ligny- Lemaire

Delivery of alpha-conotoxins Conantokin probes of the to the central nervous system human NMDA receptor Nicotinic acetylcholine receptors are The NMDA-glutamate ion channel important members of the ligand- is an abundant excitatory amino gated ion channel superfamily that acid receptor in the brain. Recent are intimately involved in signal results from our group have identified transmission. The goal of our conantokin-G analogues which research is to develop alpha- differentiate between NMDA conotoxins that can act centrally receptors found in different regions as subtype-selective inhibitors of of normal and Alzheimer human nAChRs. Our approach is to design, brain. The aim of this project is synthesise and evaluate a CNS to understand the molecular basis delivery system for alpha-conotoxins for this selectivity, with the view conjugated to carrier molecules. to better understanding and perhaps treating Alzheimer’s disease. 66 Program in Biological Chemistry 67

Combinatorial ch emistry and molecular design

We are developing Conformational diversity of the chemical and biological universes proteins and focusing the resource-intensive methods to discover Protein structure is currently combinatorial chemistry process on small molecule classified by the shape of the molecules that mimic protein shapes. polymeric backbone. Unfortunately protein mimetics, by this provides medicinal chemists New linkers for library studying the chemical with very little information as to synthesis and conformational We have developed a backbone linker and a diversity of protein safety catch linker for surfaces. This the synthesis of libraries of macrocycles (cyclic information, together molecules like chlorophyll with and cyclosporin). In chemoinformatics particular we have used the safety catch linker to and the development synthesise large arrays of peptidic macrocycles. We of new linkers and the shape or topography of amino are optimising chemistries for the acid side-chains on protein surfaces. cyclisation solid phase synthesis of large arrays We have developed the required of macrocycles containing unnatural auxiliaries, allows the algorithms and applied these to polymers and amide bond isosteres synthesis of arrays cluster continuous and discontinuous on these linkers. We are developing protein surfaces. These privileged of molecules that new linkers for the synthesis of side-chain shapes serve as screens in macrocycles. mimic protein surface an in silico assay to identify small shapes. Our work is molecules that match these shapes in Overcoming the difficult aimed at meeting the a virtual screening of virtual library macrocyclisation step approach. We have developed a novel ring huge pharmaceutical contraction auxiliary (the formation Virtual screening of virtual of a larger ring designed to undergo demands for small libraries a ring contraction and form a molecules that We are developing databases of smaller ring) that, via a ring closure/ compounds that encapsulate the modulate protein ring contraction approach, yields wealth of chemical diversity monocyclic products unobtainable function. available to synthetic chemists. We using existing synthetic methods. We have developed, and are continually are examining the combination of optimising, a drug design this strategy with new and existing environment that allows us to linkers to provide the solid-phase identify compounds in the chemical avenues for the synthesis of highly universe that match the shape of strained and novel macrocycles. proteins. In this way we are bridging 66 Program in Biological Chemistry 67

Combinatorial ch emistry

and molecular design Group leader • Mark Smythe Postdoctoral staff • Greg Bourne, Wim Meutermans, Marc Campitelli, Steve Love, Tran Trung Tran, Ailsa McManus The development of potent physiological conditions. Although Andrew McDevitt, G-protein coupled receptor the therapeutic use of cytokines Students • agonists and antagonists Doug Horton, Stephen Long, is still at an early stage, early Mikael Kofod Hansen Using our ring contraction auxiliary, indications suggest that cytokines we have synthesised a class of may be used against cancer, hepatitis macrocycles that are unobtainable Research assistant • C, multiple sclerosis, growth, asthma Simon Golding using existing chemistries. This class and arthritis. Several cytokines of natural products have shown currently on the market have wide-ranging biological activities achieved sales well in excess of

including antitumour, herbicide, $1 billion/year. Considerable effort antimalarial, opiate agonist and has therefore been devoted to the tachykinin NK-2 antagonist search for small molecule agonists activities. We are exploring this class and antagonists of key cytokines, of macrocycles as orphan ligands for but unfortunately these efforts have G-protein coupled receptors. met with little or no success. We are developing the required Cytokine mimetics design and synthetic methodologies Cytokines are a broad class of for the discovery of molecules proteins, including interferons, that modulate cytokine function. interleukins, colony stimulating Using these structure-based design factors, TGF-b and tumour necrosis approaches we have developed an factor, that regulate cell-cell antagonist of Interleukin-4. interactions under both normal and 68 69

IMB associates

Matt Trau: Nanotechnology and biomaterials

Our group focuses on the formation of novel materials and devices for medical and diagnostic needs. Examples of these include devices for rapid DNA sequencing, genetic screening, and drug discovery. Other devices of interest are artificial tissue matrices for human bone, liver and pancreas tissue. Nanotechnology, the ability to control and manipulate nanometer-sized features of matter, is a fundamental tool which is used to fashion such materials and devices.

The overall goal of the research structure are just some of the within our centre is to develop important research areas requiring biologically related materials and efficient methods of screening DNA. devices that will ultimately improve We are currently developing colloidal human health. The two main research devices comprised of microscopic categories are genetic screening and particles that can be used for rapid drug discovery devices, and artificial DNA sequencing, genetic screening tissue matrices for implants into the and drug discovery applications. human body. Both of these areas require the preparation of novel Artificial tissue matrices materials and devices which have Many human ailments arise as a been fashioned to contain designed result of the body’s inability to fully nanostructure. regenerate damaged tissue (eg. bone, liver, pancreas). Common medical Genetic screening and drug therapies in these cases involve discovery devices the use of autografts (implants Rapid access to genetic information from one’s own body), allografts is central to the revolution occurring (implants from cadavers), or other in the pharmaceutical industry, synthetic (non-biological) materials, particularly in relation to novel each of which have their associated drug target identification and drug problems. Our research focuses on development. Genetic variation, gene developing novel biological, expression, gene function and gene degradable and “living” implants for the human body. 68 69 Steve Barker: Evolutionary genomics of arthropods Most animals are arthropods, yet their genomes and evolutionary genetics are poorly understood. Our research focuses on the ticks of cattle, and the lice of humans and other primates. These parasites cause disease and economic burden, and our research is aimed at novel strategies for their containment and control.

Mitochondrial genomics Mitochondria have their own genomes, and in most groups of animals the order of genes is remarkably similar. However, we discovered two remarkable exceptions: a group of hard ticks and lice and their kin. The arrangement of the 37 genes in the mitochondria of these animals has changed so many times that it is difficult to reconstruct the evolutionary path of these mitochondria. By studying the exceptions, mitochondria that have changed a lot, we hope to learn about the rule (why the arrangement of genes in the mitochondria evolve so slowly).

Nuclear genomics We have begun to sequence the genome of Rhipicephalus appendiculatus, a tick that infests cattle in Africa and transmits a malaria-like disease. R. appendiculatus will be the first chelicerate arthropod to be studied in this way, and we expect to discover much that is new.

Molecular epidemiology and evolutionary genetics of parasitic diseases We studied the epidemiology and genetics of four pathogens that cause disease: the head and body lice of humans, the scabies mite of humans and dogs, the paralysis tick of dogs and a single-celled animal called Perkinsus that kills abalone.

Bryan Mowry: Molecular genetics of schizophrenia

Schizophrenia is a brain disease characterised by hallucinations, delusional ideas, disordered speech and thinking, as well as deficits in emotional and social behaviour. It is now generally accepted that schizophrenia has a predominantly genetic basis. Our work is aimed at identifying genes for this devastating disease that afflicts an estimated 1 percent of the world population.

While the causes for schizophrenia remain unknown, evidence from family, twin and adoption studies clearly demonstrate that it aggregates in families, with this clustering largely attributable to genetic rather than cultural or environmental factors. However, epidemiological data and molecular genetic studies demonstrate that susceptibility to schizophrenia is not the result of a mutation to a single major gene. Rather, it is likely there is a number of interacting genes of small to moderate effect that interact with each other and with non-genetic risk factors to confer schizophrenia susceptibility.

Current candidate-gene and genome-wide linkage studies provide some evidence for the involvement of a number of specific genes (eg. serotonin 5HT2a receptor gene and the dopamine D3 receptor gene) and as yet unidentified factors localised to specific chromosomal regions including 6p, 6q, 8p, 13q and 22q. These data provide suggestive, but no conclusive evidence for causative genes

Our work is aimed at identifying these genes. In collaboration with national and international colleagues, we are studying both ethnically diverse (heterogeneous) and genetically isolated (homogeneous) populations, to account for two different possibilities: (1) the same, frequently occurring causative genes occur in all populations and (2) rare genes may exist in one or more genetically homogeneous populations. 70 71 Tom Loy: Molecular archaeology

Molecular archaeology provides another dimension of detail to archaeological research. It involves the analysis of DNA and proteins extracted from ancient plant and animal remains. Our areas of interest lie in the preservation of biomolecules, sex determination, species identification, population genetics, identification of diseases in the past and primate/human evolution.

Method development This year our group has developed three robust methods for the analysis of ancient DNA. We have optimised extraction and PCR amplification techniques from blood and bone, using extremely small sample sizes to minimise the impact on ancient artifacts. We have developed a sex determining test of 90% accuracy and a species determination test that provides useful phylogenetic information. Population analysis uses mitochondrial control region sequences and has been applied to ancient human populations at the Mayan site of Copan in Honduras.

Human genetic and cultural evolution This research is focusing upon blood residues on tools and bones from hominid sites in South Africa, sequence and phylogenetic analysis using the mitochondrial 16S and CytB genes, variable domains 3, 4, 5 in the 28S rRNA gene and primate endogenous retroviruses that will serve as a proxy for genomic evolution. We are also investigating ancient diseases, both infectious and hereditary, to determine their age, evolutionary development and any gene linkages. Using a combination of capillary electrophoresis, PCR amplification, decomposition studies and taphonomy we are documenting the pathways of degradation and preservation of ancient biomolecules.

John Hancock: Mammalian signal transduction

Ras genes encode small (21kD) GTP bound state and constitutively guanine nucleotide binding proteins activates its effector pathways. These that operate as molecular switches include: the Raf/ MEK/ MAP kinase in signal transduction pathways cascade, phosphatidylinositol-3- downstream of tyrosine kinase and kinase (PI3K) and networks of G-protein coupled receptors. Ras other Ras-related proteins including is activated by guanine nucleotide Ral, Rac, Rho, and Cdc42. The exchange factors that catalyse consequence of these events is Cells in complex organisms exchange of GTP for GDP. The stimulation of cell proliferation. must communicate with each binding of GTP to Ras induces a In order to function, Ras proteins other to make correct decisions conformational change that allows must be localized to the inner about their behaviour, whether RasGTP to activate multiple effector surface of the plasma membrane. they should divide, move, do pathways. Ras is returned to the This is achieved by the addition nothing or suicide. The inactive GDP ground state by the of a C-terminal membrane anchor information that cells use to hydrolysis of GTP hydrolysis, a in the endoplasmic reticulum. From make these decisions is reaction that is stimulated by GTPase there Ras proteins must be trafficked delivered to the outside of activating proteins (GAPs). Some to discrete regions or microdomains the cell and needs to be 25% of human tumours have point of the plasma membrane. How Ras transmitted across the cell mutations in one of their Ras genes proteins traffic to these domains and membrane to the nucleus, a which render the GTPase activity how Ras function is regulated by process referred to as signal of the mutant protein resistant to membrane interactions is a major transduction. Disruption of GAP stimulation. Oncogenic Ras focus of our work. This has major signal transduction networks is is therefore fixed in the activated relevance in the design of new anti- a key feature of cancer cells. cancer therapeutics. 70 71 Ian Frazer: Effective tumour immunotherapy

Tumours are different enough from the normal cells in the person in whom they grow that the body’s defences against infection can recognise them as foreign but, unlike infections, tumours are poorly controlled by these defences. There are many reasons for this – some relate to the body’s natural reluctance to attack itself, and this problem can be overcome through better vaccines. Our work focuses on development of such vaccines for cancer of the neck of the womb (cervical cancer).

Targeting immunotherapy to epithelial tumours We use a model in which mice are transplanted with skin transgenic for the E7 protein of HPV16 to mimic the immunobiology of cervical cancer. We observe that E7 transgenic skin is not rejected by immunocompetent animals even if immunised with E7 protein and adjuvant. However treatment of grafted animals with Listeria results in prompt graft rejection and acquisition of memory immune responses sufficient to allow subsequent rejection of further E7 transgenic grafts without further Listeria exposure. We are currently exploring the molecular and cellular mechanisms of rejection and developing surrogate markers for an effective immune response that could be applied to our clinical trials of E7 specific immunotherapy for cervical premalignancy.

Codon usage and expression of papillomavirus genes Many viruses including papillomaviruses utilise codons which are rarely used in mammalian genes. We have shown that papillomavirus capsid proteins are poorly expressed in undifferentiated epithelial cells but well expressed in their differentiated progeny, and that the block to expression in undifferentiated cells is overcome by modifying codon usage to the mammalian consensus. Current work is therefore addressing whether mismatches between codon usage and AA. tRNA availability are a more general mechanism for determining efficiency of gene expression in differentiated and undifferentiated cells.

Kevin Burrage: Computational biology

Computational biological One particular area of interest is modelling is a key tool in in the use of stochastic processes understanding biological to attempt to quantify the various phenomena. We are interested issues of uncertainty, mutation rates, in trying to quantify the error measurements etc in biological mutation rates and error models. Various stochastic models measurements, both of which are being developed that attempt are uncertainties, inherent in to provide insight into genetic biological systems. We hope to regulatory networks. This work is develop mathematical models feeding into another project based robust enough to cope with on the use of virtual reality and this uncertainty, and then apply sophisticated visualization this model to genetic regulatory methodologies to understand the networks with a high degree of dynamics of regulatory networks in confidence. biology - such mathematical and visual models will be used as predictive and confirmational tools for the understanding of genomic behaviour in general. 72 73

The Australian Genome Research Facility

The Australian Genome Research Facility (AGRF), formed through the Major National Research Facilities program, provides Australia’s pre-eminent central resource for genomics researchers throughout the country and overseas. The head office and the Brisbane node are hosted by the IMB. Joint The Brisbane Division provides a wide range of DNA sequencing services to a growing number of clients. The main operational focus in the past year has been to ensure that robust protocols are in place for the various DNA sequencing service options and, where possible, automate work processes. The introduction of an internet-based sample submission process is a major advance in sample handling and was designed by our own staff.

With the new funding initiative from the National Health and Medical Research Council’s (NH&MRC) Program in Medical Genomics, designed to stimulate large-scale genomics research, the Facility is expecting a considerable increase in workload. The AGRF is uniquely positioned to provide the resources to handle the increased demand created by this Program. The continued advances in automation and a change to seven-day processing should meet this additional workload.

The Research and Technology Section was officially formed in late 1999 and operates at both the Brisbane and Melbourne nodes. The Brisbane group is focussing on single cell genotyping procedures, robotic instrumentation and high-throughput processing. The success of the single cell processing is evident with the development of a number of collaborative clinical trials in the area of pre-natal diagnostics. Similarly, advances in genetic profiling of single or low cell numbers are proving useful in archaeological identification and forensics. Continued development of the robotics workstation has produced advances in the rate and control of sample processing.

A Microarray Distribution Facility has been established at the Melbourne node. Human, mouse and yeast cDNA libraries have been purchased and will be available as microarrayed slides for individual researchers in Australia. Considerable effort has been expended in replication, validation and amplification of the clones. The development of controls is an important part of ensuring that researchers are comfortable with using the microarrays to quickly generate large amounts of data. The setting up process has taken more than 12 months and will be available for general access in 2001.

This year has seen the AGRF consolidate its processes and refine its procedures, and from this solid foundation it will continue to meet the future demand generated by the growth in genomics research. 72 73

ventures

Special Research Centre for Functional and Applied Genomics

The Special Research Centre (SRC) program. This service currently has Sean Grimmond, including provision was established at the beginning of three functional micromanipulators for clone curation and replication, 2000 with John Mattick as Director and two full time staff, and is array of clone sets and analysis. Dr and David Hume assuming the role working close to capacity. The Joanne Redburn is a key appointment of Deputy Director/Manager. The facility has established methods for with experience in many applications SRC is unique in that it brings freezing of mouse sperm and of robotics. Large mouse and human together three major areas of research embryos, and intracytoplasmic sperm cDNA sets have been replicated, strength in Computer Science and injection (ICSI). Elizabeth Williams, and the first publication-quality Information Technology, Molecular the director, recently presented a expression studies in both species Genetics and Cell Biology, and modified approach to generation of have been completed. Protein Structure and Chemistry from mouse embryo-stem cell chimaeras within the University of Queensland. at the International Mouse Genome Computational biology and data- Conference in Narita, Japan. mining Although IMB is a major participant, Two key appointments, Drs Jennifer senior staff also reside in University Protein expression facility Hallinan and Rohan Teasdale, Departments of Mathematics, Large scale expression of proteins permitted rapid advances in Computer Science and Electrical remains a limiting step in structural applications of information Engineering and the schools of biology. The protein expression technology within the SRC. Dr Molecular and Microbial Sciences facility was fortunate to recruit Ms Hallinan has developed a new neural and Biomedical Sciences. Allison McLean, who has extensive net based database for clone curation industry experience in fermentation and is working on new software for The strategy of the SRC in its technology. Three, 5- and 10-litre analysis of gene expression arrays. first year has been to establish a fermenters now operate full time Dr Teasdale is an expert in database number of core technology facilities, in the generation of high yield fed- mining, and is cooperating in several not only to eliminate bottlenecks in batch cultures of bacteria producing gene discovery projects within the the pipeline from gene discovery to recombinant proteins for subsequent SRC. Other current and future application, but also to develop new structural studies. The next phase of developments within the SRC include technologies in key areas. Each of the development is the establishment of infrastructure/technology facilities operates on a cost-recovery a downstream processing facility to development in cryoelectron basis and is available both within expedite large scale purification. microscopy, proteomics, protein- the SRC and to external groups. protein interactions and chemical Some highlights of the first year of Microarray facility libraries. The overall aim is to operation include: Gene expression information provide world-class infrastructure for provides important clues to the biotechnology research and to apply Transgenic animal service likely functions of both new and those resources efficiently to the Queensland (TASQ) known mammalian genes. The SRC generation of new drugs, bioactive Transgenic mice are a key research has established a gene expression molecules and other technology tool for many members of the SRC microarray facility, directed by Dr applications. 74 75 joint ventures...

Cooperative Research Centre for the Discovery of Genes for Common Human Diseases

The Institute is a participant in the CRC for the Discovery of Genes for Common Human Diseases (Gene CRC).

The Gene CRC was established in July 1997, bringing together a number of leading Australian human genetics groups: the Institute for Molecular Bioscience, University of Queensland; the Murdoch Children’s Research Institute, Melbourne; the Queensland Institute of Medical Research, Brisbane, and the Walter and Eliza Hall Institute of Medical Research, Melbourne. ExGenix Operations Pty Ltd is the Centre Agent and Commercial Participant of the CRC. The Menzies Centre for Population Health Research, Hobart, is an Associate Member of the CRC.

The identification and study of genes that are important in determining susceptibility to common human diseases is a major objective of the Gene CRC, and the basal cell carcinoma research conducted at the IMB (see reports of Carol Wicking and Brandon Wainwright) is one of the key research projects of the CRC.

In addition, the CRC contributes to public and school education through the joint appointment of an education officer, Angela Wallace, based at IMB.

IMB staff who participate in the Gene CRC include Professor Brandon Wainwright, who is the CRC Deputy Director and Chair of Scientific Advisory Group, Dr Carol Wicking, and several research staff, students and administrative personnel. 74 75

The establishment of the IMB as an Institute of the University of Queensland invested it with the right to enrol its own research MSc and PhD students. With this in mind, the Institute is developing a Graduate Program aimed at providing broad based career training to equip our students to tackle the scientific challenges of the 21st century. In addition to a primary research project, learning opportunities will be provided in a range of areas covering commercialisation, management, bioethics, patent law and intellectual property, and emerging technologies in the biosciences.

A Graduate Education Committee was convened to oversee the program and make recommendations to the IMB Executive. This committee has two student representatives who provide a dialogue between the student body and staff. The Institute’s commitment to graduate education resulted in the creation of a Graduate Coordinator position with the primary responsibility of handling student matters. Graduate program

In its first year the IMB enrolled 10 PhD students; Senali Abayratna, Charlotte Widberg, Nicholas Drinnan, Isabel Morrow, Anthony Cook, James Smith, Christine Wells, Johan Rosengren, Manu Trabi and Kate Irvine.

Honours students undertaking their research project within the IMB will continue to enrol through the various schools of the University, further continuing the IMB’s collaborative relationship with the Faculties. IMB staff, particularly those with joint appointments, will continue to lecture in undergraduate courses.

In 2000, PhDs where awarded to six students who began their studies in the laboratories of the then CMCB and DDD; Peter Bailey, Patricia Free, Megan Law, Ailsa McManus, David Pennisi, Edmund Sim, Aaron Smith, Joel Tyndall and Clement Waine.

The IMB actively promoted its graduate program, taking part in the University’s Postgraduate Study Information Evening. Staff and students talked to a range of individuals noting that many 2nd year students were already considering the possibility of graduate study. The IMB also hosted an open house weekend for intending graduate students aimed at attracting students from interstate to undertake PhDs at the University of Queensland. Both events were well received and actively supported by staff and students alike.

The students of the IMB actively embraced the new structure, establishing an official association of the University Union. Calling themselves SIMBA (Students of the IMB Association), this group has been very effective in drawing together the students from the two former Centres. SIMBA organised a number of events throughout the year that provided students opportunities to discuss issues pertinent to their studies and role within the IMB. They also organised social events, including a particularly successful barbecue and team sports afternoon aimed at further integrating the staff and students of both Centres. 76 77 IMBcom

IMBcom was launched by The University of Queensland in March 2000 to help commercialise outcomes from research programs of the IMB.

IMBcom’s mission is to drive an internationally competitive development and commercialisation program based on the research of the Institute. Initially IMBcom was a virtual entity within the University’s technology transfer company, UniQuest Pty Ltd, being incorporated as a separate company on 11 October 2000. The Board of IMBcom Pty Ltd comprises the University’s Senior Deputy Vice-Chancellor Professor Ted Brown; Deputy Vice-Chancellor (Research) Professor Paul Greenfield; CEO of UniSeed Dr David Evans; Dr Jane Wilson; Start-up Australia Chairman Mr Ken Roberts; and Macquarie Bank Deputy Chairman Mr Mark Johnson.

IMBcom also aims to assist the IMB to ensure that its research strengths are ultimately developed by Australian industry. This in turn will help drive Queensland’s economic growth and associated employment, which is a key requirement of the State Government’s funding of the IMB. The company intends to participate in the development of incubator facilities and seed funding arrangements to help take spin-off companies to the point where they can compete effectively for early stage venture capital in the open market. Alliances

IMBcom expects that outsourcing of R&D by pharmaceutical companies to facilitate drug design and development will provide major opportunities for research contracts with the IMB. A particular window of opportunity exists in discovery research aimed at new targets and new approaches to human genetic disease.

In the longer term, the major emphasis of IMBcom will be on establishing partnerships with companies, both pharmaceutical and biotechnology, which benefit the IMB through strengthening its financial and scientific base. The focus will be on the USA, and building on the links already established in some key centres of excellence. We expect that 60 percent of external business development and marketing will be focussed on the USA region.

Another major target for IMB R&D alliances is Asia, notably Japan, China, Korea and Taiwan, where strong relationships have already been established with the pharmaceutical and biotechnology sectors. This has been achieved with the assistance of the Queensland government offices in Tokyo, Osaka, Shanghai and Taiwan, as well as Australian Trade Commissioners. There are clear opportunities in studies of diseases important to the Asian region, and through the development of research programs linked to commercialisation processes.

IMBcom is currently negotiating with several Asian companies including Itochu in Japan and several companies from China, Korea and Singapore with interests in possible joint ventures.

Growth of the Australian biotechnology and pharmaceutical industries is also accelerating and this, along with incentives for companies to conduct R&D, including the PIIP program and new programs announced in the Innovation statement, provides additional opportunities for the IMB.

Spin-Offs

The State Government’s investment in IMB is already bearing fruit in the form of several recent biotechnology start-ups driven and staffed by ex-IMB researchers, as well as new companies currently emerging from IMB’s research.

Promics Pty Ltd was established early in 2000 to commercialise the results of novel technology developed within Associate Professor David Fairlie’s laboratory in the IMB. Venture capitalists Start-up Australia and Rothschilds Bioscience of Melbourne have invested $3 million in Promics Pty Ltd.

Promics will develop and market novel small molecules that mimic structural characteristics of proteins to make therapeutic 76 77 compounds that retain the activity of proteins but have improved stability and bio- availability. The breakthrough on which Promics is based provides a world-first class of drug candidates that shut down the activity of a protein which is produced in inflammatory disorders such as rheumatoid arthritis, acute respiratory distress syndrome, sepsis and Alzheimer’s disease.

Xenome is a public, non-listed company located in Brisbane, in which the current major shareholders are Medica Holdings Limited, a publicly listed biotechnology investment company, and UniQuest Pty Ltd, the commercialisation company of the University of Queensland.

Xenome has an exclusive license to a large portfolio of novel venom peptide compounds, and has filed International PCT applications on compounds with novel chemistry, structure and therapeutic application. Xenome’s capacity to produce new intellectual property within its own laboratories, coupled with its links to the IMB, are expected to generate significant new commercialisation opportunities.

Cytokine Mimetics Cytokine Mimetics has been established to apply a novel generic technology to the development of molecules that inhibit or mimic the action of cytokines. The technology enables the sculpting of cytokine functions onto new frameworks that have improved bioavailability characteristics when compared to the native cytokine. This process has the potential to revolutionise the treatment of many important diseases.

Genset Pacific Ltd Genset Pacific Ltd is a fully integrated genomics company engaged in providing tailored genomics information. Genset Pacific Ltd has developed unique industrial expertise in massive manufacturing of custom-made synthetic DNA fragments. Its mission is to deliver to any oligonucleotide user, anywhere in the world, the most comprehensive, reliable and flexible DNA synthesis. The company is strongly committed to bring innovative solutions to the current DNA manufacturing limitations.

Australian Genome Diagnostics Pty Ltd Australian Genome Diagnostics (AGD), a joint venture between IMBcom and The Walter and Eliza Hall Institute of Medical Research, is a highly specialised pathology laboratory serving hospital and private pathology laboratories with new and innovative diagnostic tests to aid in patient management and improve health outcomes. AGD is currently focusing on breast cancer diagnosis using BRCA1 and 2 analysis, and haematological cancers with tests such as BCRabl, gene rearrangements and translocations. AGD’s role is to improve the quality of testing rather than the development of new tests.

IP Management and Development

The market research activities of our Commercial Intelligence Agency (CIA) are used to generate technical and business packages. These packages guide development milestones and assist in the generation of marketing materials used to attract large pharmaceutical companies and venture capital investors. Working closely with scientists we have developed proposals based on existing and future IP, by conducting market appraisals, developing new strategies and matching research opportunities with investor requirements.

IMBcom is also working to bridge the gap between early stage fundamental research and commercialisation. This involves securing sources of seed funding, and assisting scientists to implement development programs that provide proof of concept and validation of the business opportunity. IMBcom has provided seed funding and helped guide the technical progress of three promising early stage programs in 2000. 78 79

Dr David Leavesley Dr Matt Trau Dr James DeVoss Hanson Centre for Cancer Department of Chemistry, The Department of Chemistry, The Research, Adelaide University of Queensland University of Queensland Innate immunity, fibrosis and Gene balls and drug balls: novel Ironing out bacteria: siderophore sclerosis - lessons from a colloidal devices for DNA biosynthesis; bacterial P450s laboratory approach sequencing and drug discovery Dr Ed Nice The Ludwig Institute for Cancer Research & The CRC for Cellular Growth Factors, Melbourne Functionally directed IMB seminars proteomics Professor H Wagner Institut für Medical Mikrobiologie, Immunologie und Hygiene, Technische Universität, München, Germany Dr Bryan Mowry Dr Jennifer Hallinan Bacterial CpG-DNA mediated Wolston Park Hospital, Wacol, Centre for Sensor Signal and signalling and activation of Queensland Information Processing, The dendritic cells Current findings in the molecular University of Queensland genetics of schizophrenia Digital image analysis for cancer Dr Helen Cooper screening Development and Neurobiology Dr Wah Chiu Unit, Walter and Eliza Hall Baylor College of Medicine, Dr Tony Weiss Institute of Medical Research, Houston, USA Department of Biochemistry, The Melbourne Structural biology of herpesvirus University of Sydney The DCC/Neogenin-netrin capsid Elastin: stretching from synthetic guidance system: a molecular genes to controlled assembly strategy for cell and axonal Professor Simon Easteal migration John Curtin School of Medical Dr Brian Key Research, Canberra Anatomical Sciences, The Dr Manuel Baca Information biology and the University of Queensland Walter and Eliza Hall Institute of integration of biomedicine in the Axon navigation in the embryonic Medical Research, Melbourne genomics era vertebrate brain Phage display: everything you wished to know but were too Professor Chris Marshall FRS Dr Denis Crane afraid to ask Institute of Cancer Research, School of Biomolecular and London UK Biomedical Science, Griffith Dr Jackie Wilce Small GTPase signalling University, Brisbane Department of Chemistry/ pathways and proliferation control Biochemistry, University of Dr Helen Blanchard Western Australia, Perth Dr Jan Slot St Vincent’s Institute of Medical Studies of a protein-DNA University Medical Center and Research, Melbourne interaction by NMR spectroscopy Institute of Biomembranes, Three-dimensional structure of and X-ray crystallography Utrecht University, Netherlands caspase-8, an initiator enzyme in Immunogold labeling of ultrathin apoptosis Dr Mike Lawrence cryosections: a morphological Biomolecular Research Institute, study on ER - Golgi transport Dr Sally-Ann Poulsen Melbourne Department of Chemistry, The Understanding active site Dr Jane Andrews University of Queensland modulation in protein families - National Institute for Medical Dynamic combinatorial chemistry: two structural examples Research, London, UK an approach to drug discovery Xenopus tropicalis: frogs with Dr Wayne Tilley glowing genetics Flinders Cancer Centre, Flinders 78 79

Medical Centre, Flinders Professor Jack Pettigrew Structural evidence for multiple University, Adelaide Vision Touch and Hearing transport mechanisms through Androgen receptor structure and Research Centre, The University the golgi function in breast and prostate of Queensland cancer Brains, biological clocks and Dr Mike Quilliam bipolar disorder Institute for Marine Biosciences, Professor André Menez National Research Council of Department of Protein Dr Chiara Zurzolo Canada, Halifax, Canada Engineering, CEA, Saclay, France Cellular and Molecular Biology, MS and LC/MS for the analysis of Animal toxins, a source of Università degli Sudi di Napoli bioactive marine natural products inspiration for drug design Federico II Naples, Italy Lipid rafts and transport of GPI- Dr Robert Cappai Professor Heinrich Betz anchored proteins in polarized Department of Pathology, Max-Planck-Institute for Brain epithelial cells University of Melbourne Research, Frankfurt, Germany proteins: structure and Glycine as central Dr Paul Pilch function in disease and health neurotransmitter: analysis of its Department of Biochemistry, receptor sites Boston University School of Professor Wayne Hall Medicine, USA Executive Director Professor Robert Capon Molecular regulation of insulin The National Drug and Alcohol School of Chemistry, The action in adipocytes Research Centre, University of University of Melbourne New South Wales, Sydney Marine bioprospecting: bioactive Dr Andrea Munsterberg Heroin use in Australia: lessons metabolites from down under the Department of Anatomy and for social policy towards sea Physiology, University of Dundee, biotechnology U.K. Professor John McAvoy Muscle development in mice Dr Bill Lot Department of Anatomy, Sir Albert Sakzewski Virus University of Sydney Dr Yumik Saga Research Centre, The University Growth factors in lens Department of Toxicology, of Queensland developmental biology and National Institute for Health, Vitamin B12-RNA interactions in pathology: key roles for FGF and Japan Hepatitis C: a novel probe of TGFß MesP1 and MesP2 are essential structure and function

Professor Ray Rodgers Department of Medicine, Flinders University, Adelaide The dynamic ovarian follicular epithelium and its associated extracellular matrix

Dr Merlin Crossley 2000 Department of Biochemistry, University of Sydney Zinc finger proteins in the control of gene expression for the development of cardiac Dr Marilyn Anderson mesoderm Department of Biochemistry, La Dr Emma Whitelaw Trobe University, Melbourne Department of Biochemistry, Professor Hiroyuki Takeda Biosynthesis and function of University of Sydney National Institute of Genetics, cyclic knotted proteins in plants Epigenetic inheritance in Japan mammals Dynamic expression of zebrafish Dr Joel Adelson hairy-related gene, her1, identifies College of Physicians and Dr Mike Eccles a molecular clock linked to teleost Surgeons, Columbia University, Cancer Genetics Laboratories somite New York, USA and Department of Biochemistry, Genomic redundency, The University of Otago, Dr Kathryn Howell development of new functional New Zealand Health Sciences Center molecules, and inventiveness in Pax2 and apoptosis: life and University of Colorado, USA higher genomes death in the developing kidney 80 81

Professor György Kéri Research, Adelaide polyprotein at the membrane Department of Medicinal Protein trafficking and signal C-prM junction. Structure- Chemistry, Semmelweis Medical transduction in intracellular function analysis by CD, NMR University, Budapest, membranes: tail-anchored and directed mutagenesis Hungary proteins and sphingosine-kinase Signal transduction therapy - new Dr Terje Dokland trends in drug research Professor Mark von Itzstein Institute of Molecular Agrobiology, Director, Centre for Biomolecular The National University of Dr Martin Banwell Science and Drug Discovery, Singapore Research School of Chemistry, Griffith University, Gold Coast Viral morphogenesis: structural Institute of Advanced Studies, The synthesis and evaluation of and biochemical approaches Australian National University, carbohydrate based probes as Canberra inhibitors of rotavirus Dr David Owen Enzyme directed synthesis of key Laboratory of Molecular Biology, drug precursors Dr Matt Cooper MRC, Cambridge, UK Department of Chemistry, A structural and functional Dr Alan Roseman University of Cambridge, UK dissection of the AP2 adaptor Laboratory of Molecular Biology, Using surface plasmon resonance complex - a pivotal player in MRC, Cambridge, UK (SPR) to screen for binders to clathrin mediated endocytosis Building atomic models of membrane receptors macromolecules using cryo Dr Thomas Preiss electron microscopy Dr Francois Penin Gene Expression Programme, Institute for Biology and EMBL, Germany Dr Binks Wattenberg Chemistry of Proteins, France Analysis of mRNA recruitment to in eukaryotic cells Molecular Cell Biology Laboratory, Mechanism of signal peptide Hanson Centre for Cancer process of Dengue viral 80 81 Collaboration

is the lifeblood of scientific

endeavour, enabling synergies and exchanges that are essential for successful research and development outcomes. IMB scientists list a total of 137 international collaborative projects active in 2000, and a further 140 within Australia. These include partnerships with many of the most prestigious research institutions in the world. Our collaborative partners include:

• European Molecular Biology Laboratory, Germany • NeuroGadgets Inc., Canada • National Institutes of Health, Bethesda, USA • Duke University Medical Centre, Chapel Hill, USA • Merck Laboratories, Philadelphia, USA • Oxford University, UK • Hong Kong University, Hong Kong • Osaka University, Japan • Baylor College of Medicine, Houston, USA • Samuel Lunenfeld Research Institute, Toronto, Canada • Washington University School of Medicine, USA • The Mount Sinai School of Medicine, New York, USA • Wistar Institute, Philadelphia, USA • Harvard Medical School, Boston, USA • Imperial College, The University of London, UK • The Hospital for Sick Children, Toronto, Canada • Centre National de la Recherche Scientifique, France • Brain Research Institute, RIKEN Japan • Garvan Institute of Medical Research, Sydney • Ludwig Cancer Research Centre, Melbourne • St Vincent’s Institute of Medical Research, Melbourne • Glaxo Wellcome, USA • Cambridge University, UK • Cornell University, New York, USA • Institute of Molecular Pathology Vienna, Austria • Massachusetts General Hospital, USA • Queensland Institute of Medical Research, Brisbane • The Walter and Eliza Hall Institute, Melbourne • Children’s Medical Research Institute, Sydney • Institute of Medical and Veterinary Science, Adelaide • University College, London, UK • Fox Chase Cancer Center, Philadelphia, USA • Howard Florey Institute, Melbourne • National Cancer Institute, Frederick, USA • Biomolecular Research Institute, Canberra • Ludwig Institute for Cancer Research, Melbourne • Scripp’s University, San Diego, USA • Case Western Reserve University, Ohio, USA • Analytica Therapeutics Inc, San Francisco, USA • University of California, San Diego, USA • NovoNordisk, Copenhagen, Denmark • Washington University, St Louis, USA • University of the Witwatersrand, South Africa • San Diego Zoological Society, USA • John Curtin School of Medical Research, Canberra • University of Edinburgh, UK • University of California, San Francisco, USA 82 83 Financial statement Statement of Operating Income and Expenditure Year Ended 31 December 2000

INCOME: Note University of Queensland (Operating Grant) 1 2,942,718 University of Queensland Research Grants 200,990 State Government 2 5,500,000 SRC Grant (Australian Research Council) 1,631,153 Australian Research Council 3 1,131,271 Clive and Vera Ramaciotti Foundation 43,546 CRC for Discovery of Genes for Common Human Diseases 220,958 Diabetes Australia Research Trust 33,409

Department of Industry Science and Technology 166,400 Human Frontiers Science Program 127,242 Glaxo Wellcome Australia 670,000 Government Employees Medical Research Fund 45,000 Juvenile Diabetes Foundation International 299,626 Mayne Bequest Foundation 60,000 The Merck Genome Research Institute 261,559 National Health and Medical Research Council 3 2,938,586 National Heart Foundation 45,000 Post Graduate Scholarships 28,209 Queensland Cancer Fund 230,072 Sylvia and Charles Viertel Charitable Foundation 165,000 Wellcome Trust 28,011 Commercial Income 1,371,664 Miscellaneous Income 415,591

TOTAL INCOME: 18,556,004

Funds brought forward from 1999 4 1,009,031

TOTAL FUNDS AVAILABLE 19,565,035

EXPENDITURE:

Salaries-Research 6,549,841 -Administration 1,090,220 -Infrastructure 541,043

Research Services 2,635,745 Education Programs 5 317,726 Administration 6 937,703 Infrastructure 7 357,436 Capital Equipment 8 2,307,116 IMBcom 984,608

TOTAL EXPENDITURE: 15,721,438

Funds carried forward to 2001: 9 3,843,597 82 83

Explanatory Notes to Statement of Income and Expenditure

1/ In-kind Contributions 6/Administration Annual Report 16,670 Figure does not include the following salaries for joint Marketing 55,714 appointments paid by other departments: Personnel Recruitment and Training 155,664 Visiting Scientists/Seminars 19,435 Department Percentage Other fees 233,844 Entertaining 7,806 S. Barker Parasitology 80 Equip Lease - Photocopiers 13,178 D. Hume Biochemistry 40 Legal Expenses 667 Microbiology 40 Postage and Freight 11,065 T. Loy Anthropology & Sociology 100 Printing and Stationery 57,715 J. Mattick Biochemistry 20 Telephone 39,516 R.Parton Microscopy & Microanalysis 10 Travel Expenses 30,478 J. Rothnagel Biochemistry 100 Sundries 39,428 B.Wainwright Biochemistry 20 Facility Costs 256,525 M.Waters Physiology & Pharmacology 100 A. Yap Physiology & Pharmacology 100 B. Kobe Biochemistry 100 7/Infrastructure 2/ State Government Funding received in advance for 2001 1,750,000 General Maintenance 1,880 Rental - Demountables 12,674 Renovations 49,036 Laundry 1,396 3/ Fellowship/Projects from Government Agencies Minor Equipment and Furniture 32,160 Animals 58,746 Australian Research Council Additional Lab Space 2,609 Computer Services 37,177 Projects 867,324 Equipment Maintenance 119,047 Fellowships 263,947 Glass Washing and Replacement 34,174 Minor Equipment 33,363 Reticulated gases, RO water and dry ice 37,497 National Health and Medical Research Council Scientific Services -78,468 Stores 16,146 Projects 2,593,522 Fellowships 345,064

4/ Funds brought forward from 1999 8/Capital Equipment

University of Queensland Operating Grant 784,151 Scientific & Computing Equipment 2,219,577 Fellowships (as approved by funding bodies) -25,633 Minor Equipment 87,539 Project Grants (as approved by funding bodies) 250,512

5/ Education Programs 9/Funds carried forward to 2001 Postgraduate scholarships 298,786 Postgraduate recruitment & training 18,941 University of Queensland Operating Grant 71,883 University of Queensland Research Grants 66,234 Post Graduate Scholarships 4,937 State Government * 1,961,398 SRC Grant 513,119 Fellowships (as approved by funding bodies) 15,685 Project Grants (as approved by funding bodies) 1,210,

* See note 2 84 85 Publications 2000 84 85

Abbenante, G., Kovacs, D. M., Leung, lacking the vascular endothelial growth Clark, C. I., Reid, R. C., McGeary, R. D. L., Craik, D. J., Tanzi, R. E. and factor-B gene (Vegfb) have smaller hearts, P., Schafer, K. and Fairlie, D. P. (2000) Fairlie, D. P. (2000) Inhibitors of beta- dysfunctional coronary vasculature and Small peptides do not inhibit human non-

amyloid formation based on the beta- impaired recovery from cardiac ischemia. pancreatic secretory phospholipase-A2 secretase cleavage site. Biochemical and Circulation Research 86: e29-e35. (type IIA). Biochemical and Biophysical Biophysical Research Communications Research Communications 274: 831-834. 268: 133-135. Bowles, J., Bullejos, M. and Koopman, P. (2000) Screening for novel mammalian Clark, S. F., Molero, J. C. and James, Andrews, K. T., Walduck, A., Kelso, sex-determining genes using expression D. E. (2000) Release of insulin receptor M. J., Fairlie, D. P., Saul, A. and cloning and microarray approaches. substrate proteins from an intracellular Parsons, P. G. (2000) Anti-malarial effect Australian Biochemist 31: 4-6. protein complex coincides with the of histone deacetylation inhibitors and development of insulin resistance. Journal mammalian tumour cytodifferentiating Bowles, J., Bullejos, M. and Koopman, of Biological Chemistry 275: 3819-3826. agents. International Journal for P. (2000) A subtractive gene expression Parasitology 30: 761-768. screen suggests a role for vanin-1 in Craik, D. J., Daly, N. L. and Waine, C. testis development in mice. Genesis 27: (2000) The cystine knot motif in toxins and Angel, N. Z., Walsh, N., Forwood, M. 124-135. implications for drug design. Toxicon 39: R., Ostrowski, M. C., Cassady, A. I. and 43-60. Hume, D. A. (2000) Transgenic mice Bowles, J., Schepers, G. and Koopman, P. that overexpress the tartrate-resistant acid (2000) Phylogenetic of the SOX family of Craik, D. J. and Scanlon, M. J. (2000) phosphatase gene exhibit an increased developmental transcription factors based Marine toxins as a starting point to rate of bone turnover. Journal of Bone and on sequence and structural indicators. drugs. In "Seafood Toxicity: Mode of Mineral Research 15: 103-110. Developmental Biology 227: 239-255. Action, Pharmacology, Physiology, and Detection". (Botana, L. M., ed.), Marcel Apolloni, A., Prior, I. A., Lindsay, Dekker Inc., New York. Basel, M., Parton, R. G. and Hancock, New York. pp. 715-740. J. F. (2000) H-ras but not K-ras traffics to the plasma membrane Craik, D. J. and Scanlon, M. J. through the exocytic pathway. (2000) Pharmaceutical Molecular and Cellular Biology 20: applications of NMR. In "Annual 2475-2487. Reports on NMR Spectroscopy". (Webb, G. A., ed.), Academic Arin, M. J., Longley, M. A., Press, London. pp. 115-174. Epstein, E. H. J., Rothnagel, J. A. and Roop, D. R. (2000) Craik, D. J. and Scanlon, M. Identification of a novel mutation J. (2000) Two, three and four in keratin 1 in a family with dimensional NMR of biomolecules. epidermolytic hyperkeratosis. In "Encyclopedia of Analytical Experimental Dermatology 9: Chemistry: Instrumentation and 16-19. Applications". (Myers, R. A., ed.), John Wiley and Sons, London. Atwood, C. S., Scarpa, R. C., pp. 1-21. Huang, X., Moir, R. D., Jones, W. D., Fairlie, D. P., Tanzi, R. E. and Croft, L., Beatson, S. A., Bush, A. I. (2000) Characterization Whitchurch, C. B., Huang, B., of copper interactions with Blakeley, R. L. and Mattick, J. alzheimer amyloid beta peptides: S. (2000) An interactive web- identification of an based Pseudomonas aeruginosa attomolar-affinity copper binding genome database: discovery of site on amyloid beta1-42. Journal new genes, pathways and of Neurochemistry 75: 1219-1233. structures. Microbiology 146: 2351-2364. Battersby, B., Bryant, D., Meutermans, W., Smythe, M. and Croft, L., Schandorff, S., Clark, Trau, M. (2000) Chemical libraries F., Burrage, K., Arctander, P. of unlimited size: ‘Colloidal and Mattick, J. (2000) ISIS, the Barcoding’ in combinatorial intron information system, reveals chemistry. Journal of the American the high frequency of alternative Chemical Society 293: 521-530. splicing in the human genome. Nature Genetics 24: 340-341. Battersby, B. J., Bryant, D., Cusack, R. M., Grøndal, L., Meutermans, W., Matthew, D., Broxton, N. M., Gehrmann, J., Miranda, Abbenante, G., Fairlie, D. P., Gahan, Smythe, M. L. and Trau, M. (2000) L., Down, J., Alewood, P. F. and L. C., Hanson, G. R. and Hambley, Toward larger chemical libraries: encoding Livett, B. (2000) Leu10 of alpha-conotoxin T. W. (2000) Conformations of cyclic with fluorescent colloids in combinatorial PnIB confers potency for the mammalian octapeptides and the influence of chemistry. Journal of the American neuronal nicotinic response. European heterocyclic ring constraints upon calcium Chemical Society 122: 2138-2139. Journal Pharmacology 390: 229-236. binding. Journal of Chemical Society Behncken, S. N., Billestrup, N., Brown, Carozzi, A., Ikonen, E., Lindsay, M. and Perkin Transaction II: 323-331. R., Amstrup, J., Conway-Campbell, B. L. Parton, R. G. (2000) Role of cholesterol Daly, N. L. and Craik, D. J. (2000) Acyclic and Waters, M. J. (2000) GH independent in organization of developing T-tubules; permutants of naturally occurring cyclic dimerization of GH receptor by a leucine analogous mechanisms for T-tubule and proteins: characterization of cystine knot zipper results in constitutive activation. caveolae biogenesis. Traffic 1: 326-341. Journal of Biological Chemistry 275: and β-sheet formation in the macrocyclic 17000-17007. Chen, S.-L., Dowhan, D. H., Hosking, polypeptide Kalata B1. Journal of B. M. and Muscat, G. E. O. (2000) Biological Chemistry 265: 19068-19075. Bellomo, D., Headrick, J. P., Silins, G. U., The steriod receptor coactivator, GRIP-1, Daly, N. L., Hoffmann, R., Otvos, L. Paterson, C. A., Thomas, P. S., Gartside, is necessary for MEF-2C dependent and Craik, D. J. (2000) Role of M., Mould, A., Cahill, M. M., Tonks, I. D., gene expression and skeletal muscle phosphorylation in the conformation of Grimmond, S. M., Townson, S., Wells, C., differentiation. Genes and Development peptides implicated in Alzheimer's Little, M., Cummings, M. C., Hayward, 14: 1209-1229. τ N. K. and Kay, G. F. (2000) Mice Disease. Biochemistry 39: 9039-9046. 86 87

Dawson, N. F., Craik, D. J., McManus, A. Gagescu, R., Demaurex, N., Parton, R. G., Journal of FASEB 15: 2393-2395. M., Reynolds, E. C., Otvos Jr, L., Tregear, Hunziker, W., Huber, L. and Gruenberg, G. W. and Wade, J. D. (2000) Chemical J. (2000) The recycling endosome of Hammet, A., Pike, B. L., Mitchelhill, K. I., synthesis, characterization and activity of madin-darby canine kidney cells is a Teh, T., Kobe, B., House, C. M., Kemp, B. RK-1, a novel a-defensin-related peptide. mildly acidic compartment rich in raft E. and Heierhorst, J. (2000) FHA domain Journal of Peptide Science 6: 19-25. components. Molecular Biology of the Cell boundaries of the Dun1p and Rad53p cell 11: 2775-2791. cycle checkpoint kinases. FEBS Letters Dwidedi, P. P., Imdahl, J. L., Kola, I., 471: 141-146. Hume, D. A. and Kay, B. K. (2000) Galve-de Rochemonteix, B., Kobayashi, Regulation of rat 25-hydroxyvitamin D3 T., Rosnoblet, C., Lindsay, M., Parton, Hansford, K., Reid, R., Clark, C., 24-hydroxylase (CYP24) gene expression: R. G., Reber, G., de Maistre, E., McGeary, R. and Fairlie, D. P. (2000) evidence for cooperation of ras-activated Wahl, D., Kruithof, E. K., Gruenberg, J. "Inhibitors of human secretory Ets transcription factors with vitamin and de Moerloose, P. (2000) Interaction phospholipases A2 from unnatural amino D receptor in 1,25-(OH)2D3-mediated of anti-phospholipid antibodies with late acids" filed as "Novel Molecules". induction. Journal of Biological Chemistry endosomes of human endothelial cells. PR1669/00. Australia 275: 47-55. Atherosclerosis, Thrombosis and Vascular Hargrave, M., James, K., Nield, K., Biology 20: 563-574. Dwivedi, P. P., Omdahl, J. L., Kola, I., Toombes, C., Georgas, K., Sullivan, T., Hume, D. A. and May, B. K. (2000) Georgas, K., Bowles, J., Yamada, T., Verzijl, H. T. F. M., Fitzpatrick, D., Oley, Regulation of rat cytochrome P450C24 Koopman, P. and Little, M. H. (2000) C. A., Little, M., De Jonghe, P., Kwon, (CYP24) gene expression. Evidence for Characterisation of Crim1 expression in J. M., Sankila, E.-M., Kremer, H., Dixon, functional cooperation of Ras-activated the developing mouse urogenital tract M. J., Timmerman, V., Yamada, T. and Ets transcription factors Koopman, P. (2000) Fine with the vitamin D receptor mapping of the neurally in 1,25-dihydroxyvitamin expressed gene SOX14 to D(3)-mediated induction. human 3q23, relative to Journal of Biological three congenital diseases. Chemistry 275: 47-55. Human Genetics 106: 432-439. Evans, T., Boonchai, W., Shanley, S., Smyth, I., Hargrave, M., James, K., Gillies, S., Georgas, K., Yamada, T. and Koopman, Wainwright, B., Chenevix- P. (2000) Sox14 maps Trench, G. and Wicking, to mouse chromosome 9 C. (2000) The spectrum and shows no mutations of Patched mutations in in the neurological mouse a collection of Australian mutants ducky, tippy and basal cell carcinomas. El-1. Mammalian Genome Human Mutation 16: 43-48. 11: 231-233. Fairlie, D. P., Tyndall, J. Hargrave, M., D. A., Reid, R. C., Wong, Karunaratne, A., Cox, L., A. K., Abbenante, G., Wood, S. A., Koopman, P. Scanlon, M. J., March, D. A. and Yamada, T. (2000) R., Bergman, D. A., Chai, The HMG box transcription C. L. L. and Burkett, B. factor gene Sox14 marks A. (2000) Conformational a novel subset of ventral selection of inhibitors and interneurons and is substrates by proteolytic regulated by Sonic enzymes: Implications for hedgehog. Developmental drug design and Biology 219: 142-153. polypeptide processing. Hargrave, M. and Journal Medicinal Koopman, P. (2000) In Chemistry 43: 1271-1281. situ hybridization of whole- Fitzgerald, J., Kennedy, D., mount embryos. Methods Liao, Y. H., Cohen, B., in Molecular Biology 123: Mattick, J. S. and 279-289. Forsayeth, J. (2000) Harris, J. M., McIntosh, Unc-50 related protein E. M. and Muscat, G. E. (URP), a mammalian O. (2000) Expression and homologue of unc-50, cytoplasmic localisation of increases neuronal deoxyuridine triphosphate nicotinic receptor pyrophosphatase encoded expression. Brain by a human endogenous Research 877: 110-123. reveals a sexually dimorphic gonadal expression pattern. Developmental retrovirus. Archives of Virology 145: Fontes, M. R. M., Teh, T. and Kobe, Dynamics 219: 582-587. 353-363. B. (2000) Structural basis of recognition Gillooly, D. J., Morrow, I. C., Lindsay, Hashiramoto, M. and James, D. E. (2000) of monopartite and bipartite nuclear Characterisation of insulin responsive localization sequences by mammalian M., Gould, R., Bryant, N. J., Gaullier, J.-M., Parton, R. G. and Stenmark, H. GLUT4 storage vesicles isolated from importin-alpha. Journal of Molecular 3T3-L1 adipocytes. Molecular and Cellular Biology 297: 1885-1896. (2000) Localisation of phosphatidylinositol 3-phosphate in yeast and mammalian Biology 20: 416-427. Fowles, L. F., Stacey, K. J., Marks, D., cells. EMBO Journal 19: 4577-4588. Haynes, D. H., Harkin, D. G., Bignold, Hamilton, J. A. and Hume, D. A. (2000) Goldring, C. E., Reveneau, S., Chantome, L. P., Hutchens, M. J., Taylor, S. M. Regulation of the urokinase plasminogen and Fairlie, D. P. (2000) Inhibition of activator gene in macrophages by A., Pance, A., Fleury, C., Hume, D. A., Sester, D., Mignotte, B. and Jeannin, C5a-induced neutrophil chemotaxis and macrophage colony-stimulating factor macrophage cytokine production in vitro (CSF-1) is dependent upon the level of J. F. (2000) Heat shock enhances transcriptional activation of the murine by a new C5a receptor antagonist. cell surface receptor. Biochemical Journal Biochemical Pharmacology 60: 729-733. 347: 313-320. inducible nitric oxide synthase gene. 86 87

Hill, J. M., Alewood, P. F. and Craik, D. J. coiling: the continuum of solenoid protein the embryo. Experimental Hematology 28: (2000) Conotoxin TVIIA, a novel peptide structures. Trends in Biochemistry 25: 601-611. from the venom of Conus Tulipa 2. Three- 509-515. dimensional solution structure. European Lincoln, D. T., Kaiser, H. E., Raju, Journal of Biochemistry 267: 4649-4657. Kolle, G., Georgas, K., Holmes, G. P., G. P. and Waters, M. J. (2000) GH Little, M. H. and Yamada, T. (2000) and colorectal carcinoma: localization of Hill, J. M., Atkins, A., Loughnan, M., Jones, CRIM1, a novel gene encoding a cysteine- receptors. In Vivo 14: 41-49. A., Adams, A., Lewis, R. J., Craik, D. rich repeat protein, is developmentally J. and Alewood, P. F. (2000) Conotoxin regulated and implicated in vertebrate Little, M., Yamada, T., Holmes, G., TVIIA, a novel peptide from the venom of CNS development and organogenesis. Georgas, K., Kolle, G. and Wilkinson, L. Conus Tulipa 1. Isolation, characterisation Mechanisms of Development 90: 181-193. (2000) Novel nucleic acid and polypeptide. and chemical synthesis. European Journal PCT/AU00/01435. Australia of Biochemistry 267: 4642-4648. Lehane, L. and Lewis, R. J. (2000) Ciguatera: recent advances but the risk Little, M. H., Carman, G. and Donaldson, Hill, M. M., Connolly, L. M., Simpson, R. remains. International Journal of Food E. (2000) Novel WT1 9 mutation J. and James, D. E. (2000) Differential Microbiology 61: 91-125. (D396>Y) in a patient with early onset protein phosphorylation in 3T3-L1 Denys Drash syndrome. Human Mutation adipocytes in response to insulin versus Leung, D., Abbenante, G. and Fairlie, D. P. 15: 389. (2000) Protease inhibitors: current status PDGF. No evidence for a Lovicu, F., Kolle, G., Yamada, T., Little, M. phosphatidylinositide 3-kinase- and future prospects. Journal Medicinal Chemistry 43: 305-341. H. and McAvoy, J. W. (2000) Expression of independent pathway in insulin signaling. Crim 1 during murine ocular development. Journal of Biological Chemistry 275: Lewis, R., Nielson, K. L., Craik, D., Mechanisms of Development 94: 261-265. 24313-24320. Loughnan, M., Adams, D., Sharpe, I., Luchin, A., Purdom, G., Murphy, K., Hume, D. A. (2000) Probability in Luchian, T., Adams, D. J., Bond, T., Thomas, L., Jones, A., Matheson, J.-L., Clark, M.-Y., Angel, N. Z., Cassady, A. transcriptional regulation and its I., Hume, D. A. and Ostrowski, M. C. implications for leukocyte differentiation Drinkwater, R., Andrews, P. R. and Alewood, P. (2000) Novel w-conotoxins (2000) The microphthalmia transcription and inducible gene expression. Blood 96: factor regulates expression 2323-2328. of the tartrate-resistant acid Ikonen, E. and Parton, R. G. phosphatase gene during (2000) Caveolins and cellular terminal differentiation of cholesterol balance. Traffic 1: osteoclasts. Journal of Bone 212-217. and Mineral Research 15: 451-460. Jennings, I. G., Cotton, R. G. and Kobe, B. (2000) MacColl, G., Novo, F., Functional analysis, using Marshall, N., Rowlinson, S., in vitro mutagenesis, of Waters, M., Goldspink, G. amnio acids located in the and Bouloux, P. (2000) phenylalanine hydroxylase Optimization of GH production active site. Archives of by muscle cells using plasmid Biochemistry and Biophysics DNA vectors. Journal of 384: 238-244. Endocrinology 165: 329-336. Jennings, I. G., Cotton, R. Maerz, A. L., Center, R. G. and Kobe, B. (2000) J., Kemp, B. E., Kobe, B. Structural interpretation of and Poumbourios, P. (2000) mutations in phenylalanine Functional implications of the hydroxylase protein aids in human T-lymphotropic virus identifying genotype- type 1 transmembrane phenotype correlations in glycoprotein helical hairpin phenylketonuria. European structure. Journal of Virology Journal of Human Genetics 74: 6614-6621. 8: 683-696. Martin, S., Millar, C. A., Lyttle, Kania-Azuma, M., Mattick, J. C. T., Meerloo, T., Marsh, S., Kaibuchi, K. and Wood, B. J., Gould, G. W. and S. A. (2000) Co-localisation James, D. E. (2000) Effects of FAM and AF-6, the of insulin on intracellular GLUT4 vesicles in adipocytes: mammalian homologues of from conus catus discriminate among evidence for a secretory mode of Drosophila faf and canoe, in mouse neuronal calcium channel subtypes. regulation. Journal of Cell Science 113: eye development. Mechanisms of Journal of Biological Chemistry 275: 3427-3438. Development 91: 383-386. 35335-35344. Martin, S., Ramm, G., Lyttle, C. T., Kelso, M. J., Hoang, H. N., Appleton, T. G. Lewis, R. J. (2000) The changing face of Meerloo, T., Stoorvogel, W. and James, D. and Fairlie, D. P. (2000) The first solution ciguatera. Toxicon 39: 97-106. structure of a single alpha helical turn. A E. (2000) Biogenesis of insulin-responsive pentapeptide a-helix stabilised by a metal Lewis, R. J. (2000) Ion channel toxins GLUT4 vesicles is independent of clip. Journal of the American Chemical and therapeutics: from cone snail venoms brefeldin a-sensitive trafficking. Traffic 1: Society 122: 10488-10489. to ciguatera. Therapeutic Drug Monitoring 652-660. 22: 61-64. Kobayashi, T., Vischer, U. M., Rosnoblet, Millar, C. A., Meerloo, T., Martin, S., C., Lebrand, C., Lindsay, M., Parton, Lewis, R. J., Molgeó, J. and Adams, D. Hickson, G. R. X., Shimwell, N. J., R. G., Kruithof, E. K. and Gruenberg, J. (2000) Ciguatera toxins: pharmacology Wakelam, M. J. O., James, D. E. and J. (2000) The tetraspanin CD63/lamps3 of toxins involved in ciguatera and Gould, G. W. (2000) Adipsin and the cycles between endocytic and secretory related fish poisonings. In "Seafood and glucose transporter GLUT4 traffic to the compartments in human endothelial cells. Freshwater Toxins". (Botana, L. M., ed.), cell surface via independent pathways in Molecular Biology of the Cell 11: Marcel Dekker, Inc. New York, USA. pp. adipocytes. Traffic 1: 141-151. 419-447. 1829-1843. Miranda, L. P. and Alewood, P. F. (2000) Kobe, B. and Kajava, A. V. (2000) When Lichanska, A. and Hume, D. A. (2000) Challenges for protein chemical synthesis protein folding is simplified to protein Origins and functions of phagocytes in in 21st century: bridging genomics and 88 89

proteomics. Biopolmers 55: 217-226. Palmer, J. S., Duffy, D. L., Box, N. F., Qiu, L., Zhang, M., Sturm, R. A., Gardiner, Aitken, J. F., O'Gorman, L. E., Green, B., Tonks, I., Kay, G. and Parsons, P. G. Miranda, L. P., Meutermans, W. D. F., A. C., Hayward, N. K., Martin, N. G. (2000) Inhibition of melanin synthesis by Smythe, M. and Alewood, P. F. (2000) and Sturm, R. A. (2000) Melanocortin-1 cystamine in human melanoma cells. The Development of an activated acyl transfer receptor polymorphisms and risk of Journal of Investigative Dermatology 114: auxiliary for hindered "difficult" peptide melanoma: is the association explained 21-27. syntheseis: increasing the applicability of solely by pigmentation phenotype? amide-backbone substitution. Journal of American Journal of Human Genetics 66: Qui, L., Gabrielli, B. F., Fairlie, D. P. and Organic Chemistry 65: 5460-5468. 176-186. Parsons, P. G. (2000) Histone deacetylase inhibitors trigger a G2 checkpoint in Montgomery, J., Ilg, T., Thompson, J. Parton, R. G., Carozzi, A. and normal cells that is defective in tumor K., Kobe, B. and Handman, E. (2000) Gustavsson, J. (2000) Caves and cells. Molecular Biology of the Cell 11: Identification and predicted structure of labyrinths; caveolae and 2069-2083. a leucine-rich repeat motif shared by transverse-tubules in skeletal muscle. Leishamania major proteophosphoglycan Protoplasma 212: 15-23. Ramm, G., Slot, J. W., James, D. E. and parasite surface antigen 2. Molecular and Stoorvogel, W. (2000) Insulin recruits Biochemistry and Parasitology 107: Pennisi, D., Bowles, J., Nagy, J., Muscat, GLUT4 from specialized VAMP2-carrying 289-295. G. and Koopman, P. (2000) Mice null for vesicles as well as from the dynamic Sox18 are viable and display a mild coat endosomal/trans-Golgi network in rate Mullock, B. M., Smith, C. W., Ihrke, G., defect. Molecular and Cellular Biology 20: adipocytes. Molecular Biology of the Cell Bright, N. A., Lindsay, M., Parkinson, E. J., 9331-9336. 11: 4079-4091. Brooks, D. A., Parton, R. G., James, D. Rauk, A., Armstrong, D. E., Luzio, J. P. and A. and Fairlie, D. P. Piper, R. C. (2000) (2000) Is oxidative Syntaxin 7 is localized damage by beta amyloid to late endosome and prion peptides compartments, mediated by hydrogen associates with vamp atom transfer from 8, and is required glycine alpha-carbon to for late methionine sulfur within endosome-lysosome beta-sheets? Journal of fusion. Molecular the American Chemistry Biology of the Cell 11: Society 122: 9761-9767. 3137-3153. Reid, R., Clark, C., Murrell, A., Campbell, Hansford, K., Stoermer, N. J. H. and Barker, M. and Fairlie, D. P. S. C. (2000) (2000) "Construction of Phylogenetic inhibitors of human analyses of the secretory rhipicephaline ticks phospholipases A2 from indicate that the unnatural amino acids" genus genus filed as "Amino Acid Rhipicephalus is Derivatives". PQ8965/00. paraphylectic. Australia Molecular Phylogenetics and Renaud, J.-P., Harris, J. Evolution 16: 1-7. M., Downes, M., Burke, L. J. and Muscat, G. Muscat, G. E. O. E. O. (2000) Structure- and Dressel, U. function analysis of the (2000) Not a minute Rev-erbA and RVR to waste. Nature ligand binding domains Medicine 6: 1216-1217. Pennisi, D., Gardner, J., Chambers, D., reveals a large hydrophobic surface that Hosking, B., Peters, J., Muscat, G., Abbott, mediates corepressor binding and a ligand Najbar, L. V., Craik, D. J., Wade, J. D. C. and Koopman, P. (2000) Mutations in cavity occupied by side chains. Molecular and McLeish, M. J. (2000) Identification Sox18 underlie cardiovascular and hair Endocrinology 14: 700-717. of initiation sites for T4 lysozyme folding follicle defects in ragged mice. Nature using CD and NMR spectroscopy of Genetics 24: 434-437. Rojo, M., Emery, G., Marjomaki, V., peptide fragments. Biochemistry 39: McDowall, A. W., Parton, R. G. and 5911-5920. Pennisi, D. J., James, K., Hosking, B., Gruenberg, J. (2000) The transmembrane Muscat, G. E. and Koopman, P. (2000) protein p23 contributes to the organization Nielsen, K. J., Schroeder, T. and Lewis, Structure, mapping, and expression of of the Golgi apparatus. Journal of Cell R. (2000) Structure-activity relationships of human SOX18. Mammalian Genome 11: Science 13: 1043-1057. w-conotoxins at N-type voltage-sensitive 1147-1149. calcium channels. Journal of Molecular Schenk, G., Guddat, L. W., Ge, Y., Recognition 13: 55-70. Piper, M., Georgas, K., Yamada, T. Carrington, L. E., Hume, D. A., Hamilton, and Little, M. (2000) Expression of the S. and de Jersey, J. (2000) Identification of Oddie, G. W., Schenk, G., Angel, N. Z., vertebrate slit gene family and their mammalian-like purple acid phosphatases Walsh, N., Guddat, L. W., de Jersey, putative receptors, the Robo genes, in the in a wide range of plants. Gene 250: J., Cassady, A. I., Hamilton, S. E. and developing murine kidney. Mechanisms of 117-25. Hume, D. A. (2000) Structure, function Development 94: 213-217. and regulation of tartrate-resistant acid Schenk, G., Korsinczky, M. L., Hume, phosphatases. Bone 27: 575-584. Poulsen, S.-A., Watson, A. A., Fairlie, D. A., Hamilton, S. and de Jersey, J. D. P. and Craik, D. J. (2000) Solution (2000) Purple acid phosphatases from Otvos , L. J., Bokonyi, K., Varga, I., structures in aqueous SDS micelles of bacteria: similarities to mammalian and Otvos, B., Hoffman, R., Ertl, H. C. J., two amyloid β-peptides Aβ(1-28) mutated plant enzymes. Gene 255: 419-424. Wade, J. D., McManus, A. M., Craik, D. J. at the α-secretase cleavage site (K16E, and Bulet, P. (2000) Insect peptides with K16F). Journal of Structural Biology 130: Schepers, G., Bullejos, M., Hosking, improved protease-resistance protect mice 142-152. B. and Koopman, P. (2000) Cloning against bacterial infection. Protein Science and characterization of the Sry-related 9: 742-749. 88 89

transcription factor gene Sox8. Nucleic mammogenesis, lactation and involution. Taylor, S. M. and Fairlie, D. P. (2000) Acids Research 28: 1473-1480. Journal of Endocrinology 166: 503-510. Regulators of the anaphylatoxin C5a. Experimental Opinion on Therapies 10: Sekiya, I., Tsuji, K., Koopman, P., Smit, D. J., Smith, A. G., Parsons, 449-458. Watanabe, H., Yamada, Y., Shinomiya, P. G., Muscat, G. E. O. and Sturm, K., Nifuji, A. and Noda, M. (2000) R. A. (2000) Domains of Brn-2 that Thomas, G. R., Costelloe, E. A., Lunn, D. SOX9 enhances aggrecan gene promoter/ mediate homodimerization and interaction P., Stacey, K. J., Delaney, S. J., Passey, enhancer activity and is upregulated by with general and melanocytic transcription R., McGlinn, E. C., McMorran, B. J., retinoic acid in a cartilage-derived cell line, factors. European Journal of Biochemistry Ahadizadeh, A., Geczy, C. L., Wainwright, TC6. Journal of Biological Chemistry 275: 267: 6413-6422. B. J. and Hume, D. A. (2000) G551D cystic 10738-10744. fibrosis mice exhibit abnormal regulation of Smith, J. L., Schaffner, A. E., Hofmeister, inflammation in lungs and macrophages. Semmler, A. B. T., Whitchurch, C. B., J. K., Hartman, M., Wei, G., Forsthoefel, Journal of Immunology 164: 3870-3877. Leech, A. J. and Mattick, J. S. (2000) D., Hume, D. A. and Ostrowski, M. C. Identification of a novel gene fimV involved (2000) Ets-2 is a target for an akt (Protein Tyndall, J. D. A., Reid, R. C., Tyssen, D. in twitching motility in Pseudomonas kinase B)/Jun N-terminal kinase signaling P., Jardine, D. K., Todd, B., Passmore, M., aeruginosa. Microbiology 146: 1321-1332. pathway in macrophages of motheaten- March, D. R., Pattenden, L. K., Bergman, viable mutant mice. Molecular and Cellular D. A., Alewood, D., Hu, S.-H., Alewood, Sester, D. P., Naik, S., Beasley, S., Biology 20: 8026-8034. P. F., Birch, C., Martin, J. L. and Hume, D. A. and Stacey, K. J. (2000) Fairlie, D. P. (2000) Synthesis, stability, Phosphorothioate backbone modification Somodevilla-Torres, M. J., Hillyard, N. antiviral activity and protease-bound modulates activation of structures of substrate- murine macrophages by mimicking constrained CpG DNA. Journal of macrocyclic inhibitors of Immunology 165: HIV-1 protease. Journal 4165-4173. of Medical Chemistry 43: Shewan, A., Marsh, B. 3495-3504. J., Melvin, D. R., Martin, Urquhart, A. J., Kennedy, S., Gould, G. W. and D., Gould, S. J. and James, D. E. (2000) The Crane, D. I. (2000) cytoplasmic carboxy Interaction of Pex5p, the terminus of the insulin Type 1 peroxisome responsive facilitative targeting signal receptor, glucose transporter, with the peroxisomal GLUT4, contains an membrane proteins endosomal sorting motif Pex14p and Pex13p. distal to its dileucine Journal of Biological signal. Journal of Chemistry 275: Biochemistry 350: 99-107 4127-4136. Shiels, I. A., Taylor, S. Whitehead, J. P., Clark, M. and Fairlie, D. P. S. 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(2000) Hume, D. A. and Stow, J. L. (2000) Chaperons 5: 14-20. Fascinating funnel-web findings. Today's Localization and post-Golgi trafficking Life Science July/August: 28-33. of tumor necrosis factor-alpha in Stacey, K. J., Sester, D. P., Sweet, M. macrophages. Journal of Interferon and J. and Hume, D. A. (2000) Macrophage Wilson, J. M., de Hoop, M., Zorzi, N., Cytokine Research 20: 427-38. activation by immunostimulatory DNA. Toh, B.-H., Dotti, C. G. and Parton, Current Opinion in Microbiology and R. G. (2000) EEA1, a tethering protein Simms, L. A., Young, J., Wicking, C. A., Immunology 247: 41-58. of the early sorting endosome, shows Meltzer, S. J., Jass, J. J. and Leggett, B. a polarized distribution in hippocampal A. (2000) The apoptotic regulatory gene, Stebel, M., Vatta, P., Ruaro, M. E., Del Sal, neurons, epithelial cells and fibroblasts. BCL10, is mutated in sporadic mismatch G., Parton, R. G. and Schneider, C. (2000) Molecular Biology of the Cell 11: repair deficient colorectal cancers. Cell The growth suppressing gas1 product is 2657-2671. Death and Differentiation 7: 236-237. a GPI-linked protein. FEBS Letters 481: 152-158. Wilson, P. J., McGlinn, E., Marsh, A., Simpson, F., Whitehead, J. P. and James, Evans, T., Arnold, J., Wright, K., Biden, K., D. E. (2000) GLUT4 - at the cross roads Strachan, A. J., Woodruff, T. M., Haaima, Young, J., Wainwright, B., Wicking, C. and between membrane trafficking and signal G., Fairlie, D. P. and Taylor, S. M. Chenevix-Trench, G. (2000) Sequence transduction. Traffic 1: 1-10. (2000) A new small molecule C5a receptor variants of DLC1 in colorectal and ovarian antagonist inhibits the reverse-passive Sinowatz, F., Schams, D., Kolle, S., tumours. Human Molecular Genetics 15: arthus reaction and endotoxic shock 156-165. Plath, A., Lincoln, D. and Waters, M. J. in Rats1. Journal of Immunology 164: (2000) Cellular localization of GH receptor 6560-6565. in the bovine mammary gland during 90 91 Staff and students

Directors Richard Lewis IMB associates Helen Blanchard Peter Andrews (Co-Director) Melissa Little Steve Barker Greg Bourne John Mattick (Co-Director) Jenny Martin Kevin Burrage Jo Bowles Ian Taylor (Deputy Director) George Muscat Ian Frazer Neil Box Rob Parton John Hancock Ross Brinkworth Group leaders Mark Ragan Tom Loy Richard Brown Paul Alewood Joe Rothnagel Bryan Mowry Nia Bryant David Craik Mark Smythe Matt Trau Monica Bullejos David Fairlie Jenny Stow Nick Campbell Sean Grimmond Rick Sturm Postdoctoral Marc Campitelli Judy Halliday Rohan Teasdale research officers Amanda Carozzi Jennifer Hallinan Brandon Wainwright John Abbenante Ian Cassady David Hume Mike Waters Denise Adams Peter Cassidy David James Carol Wicking Tracy Arakaki Chris Clarke Bostjan Kobe Toshi Yamada Paramjit Bansal Richard Clark Peter Koopman Alpha Yap Bronwyn Battersby Sharon Clark 90 91

Elaine Costelloe Bob Reid Kristy James Norelle Daly Ulrika Rova Chris Johns Sue Dobson Scott Rowlinson Seetha Karunaratne Michael Dooley Sandrine Roy Tatiana Khromykh Uwe Dressel Martin Scanlon Adrian Knight Tammy Ellis Horst Schirra Lynette Knowles Lindsay Fowles Julie Scott Annette Lane Alison Franks Wenda Shurety Cath Latham Matthew Glenn Fiona Simpson Elizabeth Leeton Lisbeth Grondahl Yogendra Singh Caroline Ligny-Lemaire Johanna Gustavsson Aaron Smith John Lock Karl Hansford Ian Smyth Marion Loughnan Murray Hargrave Kate Stacey Robert Luetterforst Jonathon Harris Martin Stoermer Dominic Lunn Roy Himes Matthew Sweet Karen Malcolm Ben Huang Tran Trung Tran Lisa Marks Lubomira Jamriska Xue-Qing Wang Tanya Martin Montse Jaumot Cynthia Whitchurch Carmel McCormack Derek Kennedy Jon Whitehead Allison McLean Dianne Keough Fiona Wylie Shane McIntosh Sarah Kleeman Jun Yan Kirstin Millard Eva Kovacs Thea Monks Gwen Lawrie Research assistants Teresa Munchow Patrick Lau Udani Abeypala Deborah Nerthey Graham Leggatt Dion Auriac John Normyle Marion Loughnan Jenny Berkman Matthew O’Brien Stephen Love Trudy Bond James Palmer Donna Mahony Darren Brown Andrew Pearson Darren March Vanessa Caig Sarah Penning Sally Martin Marc Campitelli Alisa Mei Poh Pierre-Francois Mery Tara Carton Amanda Prior Wim Meutermans Wei Chen Emily Riley Fiona McMillan Richard Clark Tara Roberts Brendan McMorran Larry Croft Jenny Rowland Juan Carlos Molero Rachael De Kluyver Ke-lin Ru Jorgan Mould Jacqui Emery Marko Salas Peter Munroe Charles Ferguson Jen Sargent Kathleen Murphy Alistair Forrest Catherine Shang Derek Nancarrow Christine Gee Rachael Shaw Annette Nicke Kylie Georgas Gary Shooter Jane Olsson Susan Gillies Shane Simonsen Julie Osbourne Simon Golding Darren Smit David Pennisi Marita Goodwin Mark Stafford Albert Pol Lucia Ha Lida Stjepkovic Ian Prior Tamarind Hamwood Trazel Teh Ying Mei Qi Allison Healy Linda Thomas Timothy Ravasi David Ireland Juliana Venturato 92 93

Suzie Verma Brooke Gardiner Rina Masadah Patricia Vietheer Susan Gillies Carney Matheson Marilyn Walters Eva Golob Dan Matthews Christine Wells Joshua Green Sean McBryde Daying Wen Kathryn Green Amy McCart Mary White Brett Hamilton Ailsa McCormack Shayama Wijedasa Dominique Hannah Andrew McDevitt Lorine Wilkinson Angus Harding Emily McGhie Elizabeth Williams Mikael Kofod Hansen Edwina McGlinn Ian Wilson Michael Haslam Ailsa McManus Louisa Windus Saleh Hasnawati Kirstin Millard Jason Wyeth Vojtech Hlinka Chris Miller Huy Hoang Kevin Miranda Students Darryl Horton Isabel Morrow Senali Abayratna Doug Horton Christine Mulford Udani Abeypala Brett Hosking Jason Mulvenna Christelle Adolphe Sheree Hughes-Stamm Anna Murrell Azita Ahadizadeh Betsy Hung Simei Ng Radiya Ali Janet Hunter Susan Nixon Chris Armishaw Lita Imperial Suzanne Nugent Chris Barnes Wendy Ingram Delvac Oceandy Daniel Barry Katherine Irvine Andrew Paterson Scott Beatson Annemieke Jansen Leonard Pattenden Jonathan Beesley Angus Johnston Anthony Percival Jennifer Bolton Peta-Jane Jones Michael Piper Trudy Bond Shannon Joseph Alisa Poh John Bowen Paul Kajewski Lavinia Proctor Linda Byrne Asanka Karunaratne Maria Quimio Sean Byrnes Sharon Katschmarsky Lotten Ragnarsson Tom Chen Michael Kelso Jonathan Read Colin Cheng Monica Kessler Ayanthi Richards Jodi Clyde-Smith Luke Kirkwood Gail Robertson Becky Conway- Gabriel Kolle Paul Rohde Campbell Michael Korsinczky Karl Rosengren Tony Cook Chiyan Lau Jenny Rowland Stephan Cronau Patrick Lau Deirdre Ryan-Knight Rodney Cusack Chris Le Hasnawati Saleh Nicholas Drinnan Andrew Leech Tedjo Sasmono Kenneth Dusza Natalie Leo Yvonne Sau Ping Julie Dutton Donmienne Leung Robert Sbaglia Melissa Edeling Pawel Listwan Goslik Schepers Tim Evans Xiao-Song Liu Tina Schroeder Victoria Francis Kelly Loffler David Sester Tricia Free John Lock Cathy Shang Juliet French Stephen Long Renfu Shao Bryan Fry Cameron Lutton Iain Sharpe Lexie Friend Janani Mahalinga-lyer Clint Sharrad 92 93

Rachael Shaw Robyn Craik Elizabeth Williams Annette Shewan Mileta Duggleby Edmund Sim Barbara Feenstra Microarray Stephen Skelton Angela Gardner Joanne Redburn Aaron Smith Carole Key Chris Johns James Smith Karen Korenromp Nikolai Sokolenko Ronda Turk IMBcom Sheree Stamm Peter Andrews Dayman Steptoe Information Peter Riddles Trina Stewart technology services Ashley Bowen Ylva Strandberg Ondrej Hlinka Michael Finney Katharina Stummeyer Nelson Marques Kellie Broderick Elida Szilagyi Anthony McSweeney Kathy Nielsen Sau-Ping Tam Danny Thomas Lisa Edwards Jennifer Taylor Kate Seib Dimitra Temelcos Technical and Doug Horton Barry Thompson laboratory services Josh Tooth Wen Hong Toh Robyn Baird David Wilson Manuela Trabi Chris Barnett Christine Morrison James Tsai Henk Faber Joel Tyndall Greg McHugh Budi Utama Charles Nelson Nicholas Valmas Lance Rathbone Andrea Verhagen David Scarce Kim Vernon Dawn Walsh Patricia Vietheer Robert Vogel Marketing and Clement Waine communications Nicole Walsh Ruth Drinkwater Yu Wan Russell Griggs Joyce Wang Tania Hudspith Mary Wang Angela Wallace Vicki Wang Tsai Wang-Wei Personnel and Stacey Wardrop postgraduate Charlotte Webb students Christine Wells Ann Day Charlotte Widberg Jodie Campbell David Wilson Barbara Clyde Takahiro Yasuda Michael Young Scientific support Karl Byriel Administration and Michael Dooley finance Allison McLean Kellie Broderick Teresa Buckley TASQ Jodie Campbell Lynette Knowles