Project acronym AFRODITE
Project Advanced Fluid Research On Drag reduction In Turbulence Experiments
Researcher (PI) Jens Henrik Mikael Fransson
Host Institution (HI) KUNGLIGA TEKNISKA HOEGSKOLAN
Call Details Starting Grant (StG), PE8, ERC-2010-StG_20091028
Summary A hot topic in today's debate on global warming is drag reduction in aeronautics. The most beneficial concept for drag reduction is to maintain the major portion of the airfoil laminar. Estimations show that the potential drag reduction can be as much as 15%, which would give a significant reduction of NOx and CO emissions in the atmosphere considering that the number of aircraft take offs, only in the EU, is over 19 million per year. An important element for successful flow control, which can lead to a reduced aerodynamic drag, is enhanced physical understanding of the transition to turbulence process.
In previous wind tunnel measurements we have shown that roughness elements can be used to sensibly delay transition to turbulence. The result is revolutionary, since the common belief has been that surface roughness causes earlier transition and in turn increases the drag, and is a proof of concept of the passive control method per se. The beauty with a passive control technique is that no external energy has to be added to the flow system in order to perform the control, instead one uses the existing energy in the flow.
In this project proposal, AFRODITE, we will take this passive control method to the next level by making it twofold, more persistent and more robust. Transition prevention is the goal rather than transition delay and the method will be extended to simultaneously control separation, which is another unwanted flow phenomenon especially during airplane take offs. AFRODITE will be a catalyst for innovative research, which will lead to a cleaner sky.
Summary
A hot topic in today's debate on global warming is drag reduction in aeronautics. The most beneficial concept for drag reduction is to maintain the major portion of the airfoil laminar. Estimations show that the potential drag reduction can be as much as 15%, which would give a significant reduction of NOx and CO emissions in the atmosphere considering that the number of aircraft take offs, only in the EU, is over 19 million per year. An important element for successful flow control, which can lead to a reduced aerodynamic drag, is enhanced physical understanding of the transition to turbulence process.
In previous wind tunnel measurements we have shown that roughness elements can be used to sensibly delay transition to turbulence. The result is revolutionary, since the common belief has been that surface roughness causes earlier transition and in turn increases the drag, and is a proof of concept of the passive control method per se. The beauty with a passive control technique is that no external energy has to be added to the flow system in order to perform the control, instead one uses the existing energy in the flow.
In this project proposal, AFRODITE, we will take this passive control method to the next level by making it twofold, more persistent and more robust. Transition prevention is the goal rather than transition delay and the method will be extended to simultaneously control separation, which is another unwanted flow phenomenon especially during airplane take offs. AFRODITE will be a catalyst for innovative research, which will lead to a cleaner sky.
Max ERC Funding
1 418 399 €
Duration
Start date: 2010-11-01, End date: 2015-10-31
Project acronym assemblyNMR
Project 3D structures of bacterial supramolecular assemblies by solid-state NMR
Researcher (PI) Adam Lange
Host Institution (HI) FORSCHUNGSVERBUND BERLIN EV
Call Details Starting Grant (StG), LS1, ERC-2013-StG
Summary Supramolecular assemblies – formed by the self-assembly of hundreds of protein subunits – are part of bacterial nanomachines involved in key cellular processes. Important examples in pathogenic bacteria are pili and type 3 secretion systems (T3SS) that mediate adhesion to host cells and injection of virulence proteins. Structure determination at atomic resolution of such assemblies by standard techniques such as X-ray crystallography or solution NMR is severely limited: Intact T3SSs or pili cannot be crystallized and are also inherently insoluble. Cryo-electron microscopy techniques have recently made it possible to obtain low- and medium-resolution models, but atomic details have not been accessible at the resolution obtained in these studies, leading sometimes to inaccurate models.
I propose to use solid-state NMR (ssNMR) to fill this knowledge-gap. I could recently show that ssNMR on in vitro preparations of Salmonella T3SS needles constitutes a powerful approach to study the structure of this virulence factor. Our integrated approach also included results from electron microscopy and modeling as well as in vivo assays (Loquet et al., Nature 2012). This is the foundation of this application. I propose to extend ssNMR methodology to tackle the structures of even larger or more complex homo-oligomeric assemblies with up to 200 residues per monomeric subunit. We will apply such techniques to address the currently unknown 3D structures of type I pili and cytoskeletal bactofilin filaments. Furthermore, I want to develop strategies to directly study assemblies in a native-like setting. As a first application, I will study the 3D structure of T3SS needles when they are complemented with intact T3SSs purified from Salmonella or Shigella. The ultimate goal of this proposal is to establish ssNMR as a generally applicable method that allows solving the currently unknown structures of bacterial supramolecular assemblies at atomic resolution.
Summary
Supramolecular assemblies – formed by the self-assembly of hundreds of protein subunits – are part of bacterial nanomachines involved in key cellular processes. Important examples in pathogenic bacteria are pili and type 3 secretion systems (T3SS) that mediate adhesion to host cells and injection of virulence proteins. Structure determination at atomic resolution of such assemblies by standard techniques such as X-ray crystallography or solution NMR is severely limited: Intact T3SSs or pili cannot be crystallized and are also inherently insoluble. Cryo-electron microscopy techniques have recently made it possible to obtain low- and medium-resolution models, but atomic details have not been accessible at the resolution obtained in these studies, leading sometimes to inaccurate models.
I propose to use solid-state NMR (ssNMR) to fill this knowledge-gap. I could recently show that ssNMR on in vitro preparations of Salmonella T3SS needles constitutes a powerful approach to study the structure of this virulence factor. Our integrated approach also included results from electron microscopy and modeling as well as in vivo assays (Loquet et al., Nature 2012). This is the foundation of this application. I propose to extend ssNMR methodology to tackle the structures of even larger or more complex homo-oligomeric assemblies with up to 200 residues per monomeric subunit. We will apply such techniques to address the currently unknown 3D structures of type I pili and cytoskeletal bactofilin filaments. Furthermore, I want to develop strategies to directly study assemblies in a native-like setting. As a first application, I will study the 3D structure of T3SS needles when they are complemented with intact T3SSs purified from Salmonella or Shigella. The ultimate goal of this proposal is to establish ssNMR as a generally applicable method that allows solving the currently unknown structures of bacterial supramolecular assemblies at atomic resolution.
Max ERC Funding
1 456 000 €
Duration
Start date: 2014-05-01, End date: 2019-04-30
Project acronym DISENTANGLE
Project Untangling the Bacterial Chromosome: Condensin's Role in Sister Chromosome Separation and its Mechanisms
Researcher (PI) Stephan Gruber
Host Institution (HI) MAX-PLANCK-GESELLSCHAFT ZUR FORDERUNG DER WISSENSCHAFTEN EV
Call Details Starting Grant (StG), LS1, ERC-2010-StG_20091118
Summary A prerequisite for chromosome segregation in all living organisms is the topological unlinking of sister DNA molecules, called DNA decatenation. Decatenation is performed by DNA topoisomerases that work by transiently breaking strand(s) in one DNA double helix and passing another double helix through the temporarily created gate. How DNA topoisomerases manage to recognize linkages between sister DNA molecules and how they promote decatenation of sister chromatids (but not catenation) is still largely unknown.
The driving hypothesis of this project is that condensin promotes chromosome decatenation by guiding the unlinking activity of DNA topoisomerases. Condensin is a member of the family of SMC (Structural Maintenance of Chromosomes) protein complexes that is conserved from bacteria to humans. It forms large, ring-like structures that bind to and organize chromosomes. Efficient separation of sister chromosomes in the bacterium B. subtilis depends on the condensin complex. However, so far the precise role of condensin in chromosome segregation and its mechanisms are unclear.
To test our hypothesis, we will establish a minichromosome in bacteria that segregates in a condensin-dependent manner and measure its decatenation in vivo in the presence and absence of condensin. We will investigate the mechanism by which condensin organizes DNA within the (mini-)chromosome using techniques like chromosome conformation capture (3C) and electron microscopy. Finally, we will attempt to reconstitute for the first time the entrapment of DNA double helices by ring-like SMC protein complexes using purified components. Our results will be pivotal for understanding the action of SMC proteins in general with important implications in the separation and segregation of chromosomes in bacteria, the shaping of mitotic chromosomes and the resolution of sister chromatids during mitosis in eukaryotes.
Summary
A prerequisite for chromosome segregation in all living organisms is the topological unlinking of sister DNA molecules, called DNA decatenation. Decatenation is performed by DNA topoisomerases that work by transiently breaking strand(s) in one DNA double helix and passing another double helix through the temporarily created gate. How DNA topoisomerases manage to recognize linkages between sister DNA molecules and how they promote decatenation of sister chromatids (but not catenation) is still largely unknown.
The driving hypothesis of this project is that condensin promotes chromosome decatenation by guiding the unlinking activity of DNA topoisomerases. Condensin is a member of the family of SMC (Structural Maintenance of Chromosomes) protein complexes that is conserved from bacteria to humans. It forms large, ring-like structures that bind to and organize chromosomes. Efficient separation of sister chromosomes in the bacterium B. subtilis depends on the condensin complex. However, so far the precise role of condensin in chromosome segregation and its mechanisms are unclear.
To test our hypothesis, we will establish a minichromosome in bacteria that segregates in a condensin-dependent manner and measure its decatenation in vivo in the presence and absence of condensin. We will investigate the mechanism by which condensin organizes DNA within the (mini-)chromosome using techniques like chromosome conformation capture (3C) and electron microscopy. Finally, we will attempt to reconstitute for the first time the entrapment of DNA double helices by ring-like SMC protein complexes using purified components. Our results will be pivotal for understanding the action of SMC proteins in general with important implications in the separation and segregation of chromosomes in bacteria, the shaping of mitotic chromosomes and the resolution of sister chromatids during mitosis in eukaryotes.
Max ERC Funding
1 376 734 €
Duration
Start date: 2010-11-01, End date: 2015-10-31
Project acronym DissectIFT
Project In vitro reconstitution and mechanistic dissection of Intraflagellar Transport in C.elegans sensory cilia
Researcher (PI) Zeynep Ökten
Host Institution (HI) TECHNISCHE UNIVERSITAET MUENCHEN
Call Details Starting Grant (StG), LS1, ERC-2013-StG
Summary Cilia are microtubule-based protrusions of the plasma membrane found on many eukaryotic cells, including most cell types of the human body. Whereas the functions of motile cilia were immediately obvious, the role of the immotile or so-called primary cilia remained largely unrecognized for many decades. Once referred to as aberrant solitary cilia with no obvious function, these ancient structures now hold the promise of revealing no less than the secrets of multicellularity and development. Even though the importance of primary cilia is now evident, molecular mechanisms underlying their assembly and function are far from being understood. The construction and maintenance of cilia relies on an ancient, universally conserved machinery termed IntraFlagellar Transport (IFT). IFT requires a multi-subunit, non-membranous protein complex assembled from more than 20 distinct subunits. At the heart of IFT are the microtubule-associated motors, -kinesin and dynein-, that continuously ferry cargo in a bi-directional fashion needed for ciliary assembly and function. To pave the way towards a molecular understanding of this fascinating organelle, we propose to employ a bottom-up approach in which we stepwise reconstitute the IFT complex from recombinantly expressed subunits of the so far best understood primary cilium from C.elegans. The structural integrity and stability of the IFT complex will be characterized using multifaceted approaches such as chemical crosslinking or thermophoresis. To mechanistically dissect the kinesin-dependent transport in vitro, we will make use of enzymatic bulk and single-molecule assays. Collectively, these results will provide a quantitative understanding of the assembly and kinesin-dependent motility of the IFT machinery. Given that cells mobilize ~600 components to build their cilia, this experimental platform will significantly streamline future efforts to identify novel cargoes and the effects of putative regulators of the IFT machinery.
Summary
Cilia are microtubule-based protrusions of the plasma membrane found on many eukaryotic cells, including most cell types of the human body. Whereas the functions of motile cilia were immediately obvious, the role of the immotile or so-called primary cilia remained largely unrecognized for many decades. Once referred to as aberrant solitary cilia with no obvious function, these ancient structures now hold the promise of revealing no less than the secrets of multicellularity and development. Even though the importance of primary cilia is now evident, molecular mechanisms underlying their assembly and function are far from being understood. The construction and maintenance of cilia relies on an ancient, universally conserved machinery termed IntraFlagellar Transport (IFT). IFT requires a multi-subunit, non-membranous protein complex assembled from more than 20 distinct subunits. At the heart of IFT are the microtubule-associated motors, -kinesin and dynein-, that continuously ferry cargo in a bi-directional fashion needed for ciliary assembly and function. To pave the way towards a molecular understanding of this fascinating organelle, we propose to employ a bottom-up approach in which we stepwise reconstitute the IFT complex from recombinantly expressed subunits of the so far best understood primary cilium from C.elegans. The structural integrity and stability of the IFT complex will be characterized using multifaceted approaches such as chemical crosslinking or thermophoresis. To mechanistically dissect the kinesin-dependent transport in vitro, we will make use of enzymatic bulk and single-molecule assays. Collectively, these results will provide a quantitative understanding of the assembly and kinesin-dependent motility of the IFT machinery. Given that cells mobilize ~600 components to build their cilia, this experimental platform will significantly streamline future efforts to identify novel cargoes and the effects of putative regulators of the IFT machinery.
Max ERC Funding
1 497 740 €
Duration
Start date: 2014-03-01, End date: 2019-02-28
Project acronym DNA ORIGAMI DEVICES
Project Single-molecule studies of protein-protein and protein-DNA interactions, enabled by DNA origami
Researcher (PI) Hendrik Dietz
Host Institution (HI) TECHNISCHE UNIVERSITAET MUENCHEN
Call Details Starting Grant (StG), LS1, ERC-2010-StG_20091118
Summary Adhesive interactions between macromolecules are ubiquitously found in biology. Regulatory processes in biology depend on temporary physical inter-biomolecular interactions whose strengths are regulated by the internal state of the cell. Obtaining quantitative insight the dynamic strength of interactions between biomolecules has remained a difficult task. Single-molecule approaches can provide detailed insight into intra-molecular interactions in biomolecules. Yet, protein-protein and protein-DNA interactions have remained largely inaccessible. We propose to enable the single-molecule study of protein and protein-DNA interactions by taking advantage of the fine positional control afforded by DNA origami to overcome critical experimental challenges. As a first case study we plan to employ the DNA origami devices to study the single-molecule mechanics protein-protein and protein-DNA interactions that are relevant in the regulation of the galactose metabolism in yeast. We also seek to take steps towards a high-throughput single-molecule protein-DNA and protein-protein interaction assay to open access to a quantitative and combinatorial study of many different inter-macromolecular interactions, as well as to study the effects exerted by additional inhibiting or activating ligands. The proposed project will open up novel opportunities for a systematic study of macromolecular interactions in biology and is likely to deepen our understanding of regulatory processes in biology. Lessons that will be learned may suggest new ways to the rational design or identification of compounds that can prevent disease-causing interactions.
Summary
Adhesive interactions between macromolecules are ubiquitously found in biology. Regulatory processes in biology depend on temporary physical inter-biomolecular interactions whose strengths are regulated by the internal state of the cell. Obtaining quantitative insight the dynamic strength of interactions between biomolecules has remained a difficult task. Single-molecule approaches can provide detailed insight into intra-molecular interactions in biomolecules. Yet, protein-protein and protein-DNA interactions have remained largely inaccessible. We propose to enable the single-molecule study of protein and protein-DNA interactions by taking advantage of the fine positional control afforded by DNA origami to overcome critical experimental challenges. As a first case study we plan to employ the DNA origami devices to study the single-molecule mechanics protein-protein and protein-DNA interactions that are relevant in the regulation of the galactose metabolism in yeast. We also seek to take steps towards a high-throughput single-molecule protein-DNA and protein-protein interaction assay to open access to a quantitative and combinatorial study of many different inter-macromolecular interactions, as well as to study the effects exerted by additional inhibiting or activating ligands. The proposed project will open up novel opportunities for a systematic study of macromolecular interactions in biology and is likely to deepen our understanding of regulatory processes in biology. Lessons that will be learned may suggest new ways to the rational design or identification of compounds that can prevent disease-causing interactions.
Max ERC Funding
1 494 216 €
Duration
Start date: 2010-11-01, End date: 2015-10-31
Project acronym DNAMETRY
Project DNA based nanometry: Exploring chromatin structure and molecular motors
Researcher (PI) Ralf Seidel
Host Institution (HI) UNIVERSITAET LEIPZIG
Call Details Starting Grant (StG), LS1, ERC-2010-StG_20091118
Summary DNA metabolism is governed by a delicate balance between compacting the stored genetic information while simultaneously ensuring a highly dynamically access to it. This interdisciplinary project aims (i) to understand the mechanics and dynamics of chromatin as well as the mechanism of enzymes involved in DNA metabolism on a molecular level and (ii) to develop new nanometric tools based on optical methods and 3D DNA nanostructures that allow addressing new experimental questions. Within the research project novel nanoscopic detection assays based on the combination of magnetic tweezers and optical methods shall be developed, such as ultra-fast torque spectroscopy and combined FRET-force spectroscopy. Our single-molecule assays shall be applied to study the material properties of self-assembled 3D DNA nanostructures, which shall then be used to set up improved high resolution single-molecule assays. These technological improvements will become key to obtain insight into structure and dynamics of in vitro reconstituted chromatin as response to external mechanical stress but also into the operation of molecular motors that themselves generate forces and torques on DNA and chromatin. The main goal of the project is to use nanotechnological tools to understand design principles of biomolecules, biomaterials and biological motors, which in turn shall be used to develop smarter nanotools and functional elements.
Summary
DNA metabolism is governed by a delicate balance between compacting the stored genetic information while simultaneously ensuring a highly dynamically access to it. This interdisciplinary project aims (i) to understand the mechanics and dynamics of chromatin as well as the mechanism of enzymes involved in DNA metabolism on a molecular level and (ii) to develop new nanometric tools based on optical methods and 3D DNA nanostructures that allow addressing new experimental questions. Within the research project novel nanoscopic detection assays based on the combination of magnetic tweezers and optical methods shall be developed, such as ultra-fast torque spectroscopy and combined FRET-force spectroscopy. Our single-molecule assays shall be applied to study the material properties of self-assembled 3D DNA nanostructures, which shall then be used to set up improved high resolution single-molecule assays. These technological improvements will become key to obtain insight into structure and dynamics of in vitro reconstituted chromatin as response to external mechanical stress but also into the operation of molecular motors that themselves generate forces and torques on DNA and chromatin. The main goal of the project is to use nanotechnological tools to understand design principles of biomolecules, biomaterials and biological motors, which in turn shall be used to develop smarter nanotools and functional elements.
Max ERC Funding
1 500 000 €
Duration
Start date: 2011-01-01, End date: 2016-10-31
Project acronym DROPCELLARRAY
Project DropletMicroarrays: Ultra High-Throughput Screening of Cells in 3D Microenvironments
Researcher (PI) Pavel Levkin
Host Institution (HI) KARLSRUHER INSTITUT FUER TECHNOLOGIE
Call Details Starting Grant (StG), PE8, ERC-2013-StG
Summary High-throughput (HT) screening of live cells is crucial to accelerate both fundamental biological research and discovery of new drugs. Current methods for HT cell screenings, however, either require a large number of microplates, are prone to cross-contaminations and are limited to adherent cells (cell microarrays), or are not compatible with adherent cells as well as with spatial indexing (droplet microfluidics). We recently demonstrated the use of superhydrophobic-superhydrophilic microarrays to create high-density arrays of microdroplets or hydrogel micropads. We propose here to develop a new platform for HT cell screening experiments using the unique properties of the superhydrophilic microarrays separated by superhydrophobic thin barriers. The new technology will allow us to perform up to 300K cell experiments in parallel using a single chip. Individual cell experiments will be performed in thousands of completely isolated microdroplet at defined locations on the chip. This will enable spatial indexing, time-lapse measurements and screening of either adherent or non-adherent cells. Parallel manipulations within individual microreservoirs, such as washing, addition of chemical libraries, or staining will be developed to open new possibilities in the field of live cell studies. Superhydrophobic barriers will allow complete isolation of the microreservoirs, thus preventing cross-contamination and cell migration. We will also develop a technology for the HT screening of cells in 3D hydrogel micropads. We will use these methods to gain better understanding of how different parameters of the 3D cell microenvironment influence various aspects of cell behavior. The project will require the development of new technological tools which can later be applied to a wide range of cell screening experiments and biological problems. Our long term aim is to replace the outdated microplate technology with a more powerful and convenient method for cell screening experiments.
Summary
High-throughput (HT) screening of live cells is crucial to accelerate both fundamental biological research and discovery of new drugs. Current methods for HT cell screenings, however, either require a large number of microplates, are prone to cross-contaminations and are limited to adherent cells (cell microarrays), or are not compatible with adherent cells as well as with spatial indexing (droplet microfluidics). We recently demonstrated the use of superhydrophobic-superhydrophilic microarrays to create high-density arrays of microdroplets or hydrogel micropads. We propose here to develop a new platform for HT cell screening experiments using the unique properties of the superhydrophilic microarrays separated by superhydrophobic thin barriers. The new technology will allow us to perform up to 300K cell experiments in parallel using a single chip. Individual cell experiments will be performed in thousands of completely isolated microdroplet at defined locations on the chip. This will enable spatial indexing, time-lapse measurements and screening of either adherent or non-adherent cells. Parallel manipulations within individual microreservoirs, such as washing, addition of chemical libraries, or staining will be developed to open new possibilities in the field of live cell studies. Superhydrophobic barriers will allow complete isolation of the microreservoirs, thus preventing cross-contamination and cell migration. We will also develop a technology for the HT screening of cells in 3D hydrogel micropads. We will use these methods to gain better understanding of how different parameters of the 3D cell microenvironment influence various aspects of cell behavior. The project will require the development of new technological tools which can later be applied to a wide range of cell screening experiments and biological problems. Our long term aim is to replace the outdated microplate technology with a more powerful and convenient method for cell screening experiments.
Max ERC Funding
1 499 820 €
Duration
Start date: 2014-02-01, End date: 2019-01-31
Project acronym GAMETE RECOGNITION
Project Molecular Basis of Mammalian Egg-Sperm Interaction
Researcher (PI) Luca Vincenzo Luigi Jovine
Host Institution (HI) KAROLINSKA INSTITUTET
Call Details Starting Grant (StG), LS1, ERC-2010-StG_20091118
Summary At the dawn of the 21st century, our knowledge of the molecular mechanism of mammalian
fertilization remains very limited. Different lines of evidence indicate that initial gamete recognition
depends on interaction between a few distinct proteins on sperm and ZP3, a major component of the
extracellular coat of oocytes, the zona pellucida (ZP). On the other hand, recent findings suggest an
alternative mechanism in which cleavage of another ZP subunit, ZP2, regulates binding of gametes
by altering the global structure of the ZP. Progress in the field has been hindered by the paucity and
heterogeneity of native egg-sperm recognition proteins, so that novel approaches are needed to
reconcile all available data into a single consistent model of fertilization. Following our recent
determination of the structure of the most conserved domain of sperm receptor ZP3 by X-ray
crystallography, we will conclusively establish the basis of mammalian gamete recognition by
performing structural studies of homogeneous, biologically active recombinant proteins. First, we
will combine crystallographic studies of isolated ZP subunits with electron microscopy analysis of
their filaments to build a structural model of the ZP. Second, structures of key egg-sperm
recognition protein complexes will be determined. Third, we will investigate how proteolysis of
ZP2 triggers overall conformational changes of the ZP upon gamete fusion. Together with
functional analysis of mutant proteins, these studies will provide atomic resolution snapshots of the
most crucial step in the beginning of a new life, directly visualizing molecular determinants
responsible for species-restricted gamete interaction at fertilization. The progressive decrease of
births in the Western world and inadequacy of current contraceptive methods in developing
countries underscore an urgent need for a modern approach to reproductive welfare. This research
will not only shed light on a truly fundamental biological problem, but also constitute a solid
foundation for the reproductive medicine of the future.
Summary
At the dawn of the 21st century, our knowledge of the molecular mechanism of mammalian
fertilization remains very limited. Different lines of evidence indicate that initial gamete recognition
depends on interaction between a few distinct proteins on sperm and ZP3, a major component of the
extracellular coat of oocytes, the zona pellucida (ZP). On the other hand, recent findings suggest an
alternative mechanism in which cleavage of another ZP subunit, ZP2, regulates binding of gametes
by altering the global structure of the ZP. Progress in the field has been hindered by the paucity and
heterogeneity of native egg-sperm recognition proteins, so that novel approaches are needed to
reconcile all available data into a single consistent model of fertilization. Following our recent
determination of the structure of the most conserved domain of sperm receptor ZP3 by X-ray
crystallography, we will conclusively establish the basis of mammalian gamete recognition by
performing structural studies of homogeneous, biologically active recombinant proteins. First, we
will combine crystallographic studies of isolated ZP subunits with electron microscopy analysis of
their filaments to build a structural model of the ZP. Second, structures of key egg-sperm
recognition protein complexes will be determined. Third, we will investigate how proteolysis of
ZP2 triggers overall conformational changes of the ZP upon gamete fusion. Together with
functional analysis of mutant proteins, these studies will provide atomic resolution snapshots of the
most crucial step in the beginning of a new life, directly visualizing molecular determinants
responsible for species-restricted gamete interaction at fertilization. The progressive decrease of
births in the Western world and inadequacy of current contraceptive methods in developing
countries underscore an urgent need for a modern approach to reproductive welfare. This research
will not only shed light on a truly fundamental biological problem, but also constitute a solid
foundation for the reproductive medicine of the future.
Max ERC Funding
1 499 282 €
Duration
Start date: 2011-01-01, End date: 2015-12-31
Project acronym MULTIMATE
Project A Research Platform Addressing Outstanding Research Challenges for Nanoscale Design and Engineering of Multifunctional Material
Researcher (PI) Johanna Rosen
Host Institution (HI) LINKOPINGS UNIVERSITET
Call Details Starting Grant (StG), PE8, ERC-2010-StG_20091028
Summary "Nanoscale engineering is a fascinating research field spawning extraordinary materials which revolutionize microelectronics, medicine,energy production, etc. Still, there is a need for new materials and synthesis methods to offer unprecedented properties for use in future applications.
In this research project, I will conduct fundamental science investigations focused towards the development of novel materials with tailor-made properties, achieved by precise control of the materials structure and compostition. The objectives are to: 1) Perform novel synthesis of graphene. 2) Explore nanoscale engineering of ""graphene-based"" materials, based on more than one atomic element. 3) Tailor uniquely combined metallic/ceramic/magnetic materials properties in so called MAX phases. 4) Provide proof of concept for thin film architectures in advanced applications that require specific mechanical, tribological, electronic, and magnetic properties.
This initative involves advanced materials design by a new and unique synthesis method based on cathodic arc. Research breakthroughs are envisioned: Functionalized graphene-based and fullerene-like compounds are expected to have a major impact on tribology and electronic applications. The MAX phases are expected to be a new candidate for applications within low friction contacts, electronics, as well as spintronics. In particular, single crystal devices are predicted through tuning of tunnel magnetoresistance (TMR) and anisotropic conductivity (from insulating to n-and p-type).
I can lead this innovative and interdisciplinary project, with a unique background combining relevant research areas: arc process development, plasma processing, materials synthesis and engineering, characterization, along with theory and modelling."
Summary
"Nanoscale engineering is a fascinating research field spawning extraordinary materials which revolutionize microelectronics, medicine,energy production, etc. Still, there is a need for new materials and synthesis methods to offer unprecedented properties for use in future applications.
In this research project, I will conduct fundamental science investigations focused towards the development of novel materials with tailor-made properties, achieved by precise control of the materials structure and compostition. The objectives are to: 1) Perform novel synthesis of graphene. 2) Explore nanoscale engineering of ""graphene-based"" materials, based on more than one atomic element. 3) Tailor uniquely combined metallic/ceramic/magnetic materials properties in so called MAX phases. 4) Provide proof of concept for thin film architectures in advanced applications that require specific mechanical, tribological, electronic, and magnetic properties.
This initative involves advanced materials design by a new and unique synthesis method based on cathodic arc. Research breakthroughs are envisioned: Functionalized graphene-based and fullerene-like compounds are expected to have a major impact on tribology and electronic applications. The MAX phases are expected to be a new candidate for applications within low friction contacts, electronics, as well as spintronics. In particular, single crystal devices are predicted through tuning of tunnel magnetoresistance (TMR) and anisotropic conductivity (from insulating to n-and p-type).
I can lead this innovative and interdisciplinary project, with a unique background combining relevant research areas: arc process development, plasma processing, materials synthesis and engineering, characterization, along with theory and modelling."
Max ERC Funding
1 484 700 €
Duration
Start date: 2010-09-01, End date: 2015-08-31
Project acronym PISILENCE
Project Small RNA-guided complex machinery for epigenetic silencing
Researcher (PI) Ramesh Shanmughom Pillai
Host Institution (HI) EUROPEAN MOLECULAR BIOLOGY LABORATORY
Call Details Starting Grant (StG), LS1, ERC-2010-StG_20091118
Summary Transposons are parasitic DNA elements that when activated can insert into new genomic locations, leading to genome
instability. Animal germlines express a special class of small RNAs called piwi-interacting RNAs (piRNAs) which are
implicated in transposon silencing. In mammals, they are believed to promote DNA methylation of transposon-rich genomic
regions. Mechanisms of piRNA biogenesis and function are only beginning to be understood. Sequence analysis of piRNAs
from a variety of organisms has given rise to working models for piRNA biogenesis and how piRNAs might act as guides
for nuclear silencing complexes. However, key components of the pathway, especially those for biogenesis and function,
remain to be discovered. Our aim is to identify and characterize new components using a combination of biochemistry, mouse
genetics and small RNA bioinformatics. The regulatory potential of posttranslational modification of piwi proteins and their
recognition by tudor proteins will be examined. Mouse mutants will be used to study the in vivo role of catalytic activity of
piwi proteins in piRNA biogenesis and identify putative targets by transcriptomics approaches. Integrated biochemical and
deep-sequencing methods will be applied to understand how small RNAs might guide nulcear silencing machinery to target
locations. Finally, in a field that is dependent on model organisms, we propose to develop a cell culture system to study the
piRNA pathway and carry out a high-throughput functional RNAi screen for component discovery. This proposal aims to
use interdisciplinary approaches in uncovering the biochemical framework in which germline small RNAs function to protect
eukaryotic genomes.
Summary
Transposons are parasitic DNA elements that when activated can insert into new genomic locations, leading to genome
instability. Animal germlines express a special class of small RNAs called piwi-interacting RNAs (piRNAs) which are
implicated in transposon silencing. In mammals, they are believed to promote DNA methylation of transposon-rich genomic
regions. Mechanisms of piRNA biogenesis and function are only beginning to be understood. Sequence analysis of piRNAs
from a variety of organisms has given rise to working models for piRNA biogenesis and how piRNAs might act as guides
for nuclear silencing complexes. However, key components of the pathway, especially those for biogenesis and function,
remain to be discovered. Our aim is to identify and characterize new components using a combination of biochemistry, mouse
genetics and small RNA bioinformatics. The regulatory potential of posttranslational modification of piwi proteins and their
recognition by tudor proteins will be examined. Mouse mutants will be used to study the in vivo role of catalytic activity of
piwi proteins in piRNA biogenesis and identify putative targets by transcriptomics approaches. Integrated biochemical and
deep-sequencing methods will be applied to understand how small RNAs might guide nulcear silencing machinery to target
locations. Finally, in a field that is dependent on model organisms, we propose to develop a cell culture system to study the
piRNA pathway and carry out a high-throughput functional RNAi screen for component discovery. This proposal aims to
use interdisciplinary approaches in uncovering the biochemical framework in which germline small RNAs function to protect
eukaryotic genomes.
Max ERC Funding
902 849 €
Duration
Start date: 2011-01-01, End date: 2016-05-31
