ERC FUNDED PROJECTS

For over half a millennium, the great medieval capital of Angkor lay at the heart of a vast empire stretching across much of mainland SE Asia. Recent research has revealed that the famous monuments of Angkor were merely the epicentre of an immense settlement complex, with highly elaborate engineering works designed to manage water and mitigate the uncertainty of monsoon rains. Compelling evidence is now emerging that other temple complexes of the medieval Khmer Empire may also have formed the urban cores of dispersed, low-density settlements with similar systems of hydraulic engineering. Using innovative airborne laser scanning (‘lidar’) technology, CALI will uncover, map and compare archaeological landscapes around all the major temple complexes of Cambodia, with a view to understanding what role these complex and vulnerable water management schemes played in the growth and decline of early civilisations in SE Asia. CALI will evaluate the hypothesis that the Khmer civilisation, in a bid to overcome the inherent constraints of a monsoon environment, became locked into rigid and inflexible traditions of urban development and large-scale hydraulic engineering that constrained their ability to adapt to rapidly-changing social, political and environmental circumstances. By integrating data and techniques from fast-developing archaeological sciences like remote sensing, palaeoclimatology and geoinformatics, this work will provide important insights into the reasons for the collapse of inland agrarian empires in the middle of the second millennium AD, a transition that marks the emergence of modern mainland SE Asia. The lidar data will provide a comprehensive and internally-consistent archive of urban form at a regional scale, and offer a unique experimental space for evaluating socio-ecological resilience, persistence and transformation over two thousand years of human history, with clear implications for our understanding of contemporary urbanism and of urban futures.

1 482 844 €

Start date: 2015-03-01, End date: 2020-02-29

Signal transduction in biology relies on the transfer of information across biomolecules by concerted conformational changes that cannot currently be characterized experimentally at high resolution. In CONCERT we will develop a method based on the use of nuclear magnetic resonance spectroscopy in solution that will provide very detailed descriptions of such changes by using the information about structural heterogeneity contained in a parameter that is exquisitely sensitive to molecular shape called residual dipolar coupling measured in steric alignment. To show how this new method will allow the study of information transfer we will determine conformational ensembles that will report on the intra and inter-domain concerted conformational changes that activate the androgen receptor, a large allosteric multi-domain protein that regulates the male phenotype and is a therapeutic target for castration resistant prostate cancer, the condition suffered by prostate cancer patients that have become refractory to hormone therapy, the first line of treatment for this disease. To complement the structural information obtained by nuclear magnetic resonance and, especially, measure the rate of information transfer across the androgen receptor we will carry out in a collaborative fashion high precision single molecule Förster resonance energy transfer and fluorescence correlation spectroscopy experiments on AR constructs labelled with fluorescent dyes. In summary we will develop a method that will make it possible to describe some of the most fascinating biological phenomena, such as allostery and signal transduction, and will, in the long term, be an instrument for the discovery of drugs to treat castration resistant prostate cancer, a late stage of prostate cancer that is incurable and kills ca. 70.000 European men every year.

1 950 000 €

Start date: 2015-07-01, End date: 2020-06-30

Chromatin assembly is a fundamental cellular process necessary for the maintenance of genome integrity and transcriptional programs. Understanding the effect of DNA replication on histone protein dynamics is also a prerequisite for understanding the role of chromatin in epigenetic inheritance. Epigenetic phenomena are thought to influence cellular differentiation and cancer formation, as well as the impact of environmental factors on early development and later predispositions to disease. While epigenetic inheritance of chromatin components is, in theory, accepted as the driver of such phenomena, chromatin state inheritance per se has only been demonstrated for a few specific cases. Not much is known about histone “inheritance” beyond the facts that bulk maternal histones distribute equally among the daughter strands and are diluted two-fold after replication with newly synthesized “unmarked” histones, and that the majority of H3/H4 tetramers do not split before reassembly. We have shown previously that maternal nucleosomes stay on average within 400bp of their original binding site, implying that any potentially heritable chromatin encoded information, has to be inherited in ~1kb blocs, as smaller nucleosome domains would rapidly be diluted by new nucleosomes. I propose to develop high throughput systems for directly measuring movements of histones and chromatin regulators during genomic replication in S.cerevisiae to determine, how chromatin states survive the perturbations associated with replication. We will determine locus specific differences in the spread of maternal nucleosomes after replication, the effects of leading and lagging strand replication on nucleosome positioning and maternal nucleosome distribution, the renewal dynamics of posttranslational histone marks and chromatin binding proteins, and the kinetics of chromatin footprint re-establishment and gene (re)activation.

1 984 677 €

Start date: 2015-06-01, End date: 2020-05-31

Integral membrane metalloproteins are involved in the transport and homeostasis of metal ions, as well as in key redox reactions that have a tremendous impact on many fields within life sciences, environment, energy, and industry. Most of our understanding of fine details of biochemical processes derives from atomic or molecular structures obtained by diffraction methods on single crystal samples. However, in the case of integral membrane systems, single crystals large enough for X-ray diffraction cannot be easily obtained, and the problem of structure elucidation is largely unsolved. We have recently pioneered a breakthrough approach using Magic-Angle Spinning Nuclear Magnetic Resonance (MAS-NMR) for the atomic-level characterization of paramagnetic materials and complex biological macromolecules. The proposed project aims to leverage these new advances through a series of new concepts i) to improve the resolution and sensitivity of MAS-NMR from nuclei surrounding a paramagnetic metal ion, such as e.g. cobalt, nickel and iron, and ii) to extend its applicability to large integral membrane proteins in lipid membrane environments. With these methods, we will enable the determination of structure-activity relationships in integral membrane metalloenzymes and transporters, by combining the calculation of global structure and dynamics with measurement of the electronic features of metal ions. These goals require a leap forward with respect to today’s protocols, and we propose to achieve this through a combination of innovative NMR experiments and isotopic labeling, faster MAS rates and high magnetic fields. As outlined here, the approaches go well beyond the frontier of current research. The project will yield a broadly applicable method for the structural characterization of essential cellular processes and thereby will provide a powerful tool to solve challenges at the forefront of molecular and chemical sciences today.

2 499 375 €

Start date: 2015-09-01, End date: 2020-08-31