Project acronym EDIP
Project Evolution of Development In Plants
Researcher (PI) Jane Alison Langdale
Host Institution (HI) THE CHANCELLOR, MASTERS AND SCHOLARS OF THE UNIVERSITY OF OXFORD
Call Details Advanced Grant (AdG), LS3, ERC-2008-AdG
Summary Different morphologies evolve in different organisms in response to changing environments. As land plants evolved, developmental mechanisms were either generated de novo, or were recruited from existing toolkits and adapted to facilitate changes in form. Some of these changes occurred once, others on multiple occasions, and others were gained and then subsequently lost in a subset of lineages. Why have certain forms survived and others not? Why does a fern look different from a flowering plant, and why should developmental biologists care? By determining how many different ways there are to generate a particular morphology, we gain an understanding of whether a particular transition is constrained. This basic information allows an assessment of the extent to which genetic variation can modify developmental mechanisms and an indication of the degree of developmental plasticity that is possible and/or tolerated both within and between species. This proposal aims to characterize the developmental mechanisms that underpin the diverse shoot forms seen in extant plant species. The main goal is to compare developmental mechanisms that operate in vegetative shoots of bryophytes, lycophytes, ferns and angiosperms, with a view to understanding the constraints that limit morphological variation. Specifically, we will investigate the developmental basis of three major innovations that altered the morphology of vegetative shoots during land plant evolution: 1) formation of a multi-cellular embryo; 2) organization of apical growth centres and 3) patterning of leaves in distinct spatial arrangements along the shoot. To facilitate progress we also aim to develop transgenic methods, create mutant populations and generate digital transcriptomes for model species at key phylogenetic nodes. The proposed work will generate scenarios to explain how land plant form evolved and perhaps more importantly, how it could change in the future.
Summary
Different morphologies evolve in different organisms in response to changing environments. As land plants evolved, developmental mechanisms were either generated de novo, or were recruited from existing toolkits and adapted to facilitate changes in form. Some of these changes occurred once, others on multiple occasions, and others were gained and then subsequently lost in a subset of lineages. Why have certain forms survived and others not? Why does a fern look different from a flowering plant, and why should developmental biologists care? By determining how many different ways there are to generate a particular morphology, we gain an understanding of whether a particular transition is constrained. This basic information allows an assessment of the extent to which genetic variation can modify developmental mechanisms and an indication of the degree of developmental plasticity that is possible and/or tolerated both within and between species. This proposal aims to characterize the developmental mechanisms that underpin the diverse shoot forms seen in extant plant species. The main goal is to compare developmental mechanisms that operate in vegetative shoots of bryophytes, lycophytes, ferns and angiosperms, with a view to understanding the constraints that limit morphological variation. Specifically, we will investigate the developmental basis of three major innovations that altered the morphology of vegetative shoots during land plant evolution: 1) formation of a multi-cellular embryo; 2) organization of apical growth centres and 3) patterning of leaves in distinct spatial arrangements along the shoot. To facilitate progress we also aim to develop transgenic methods, create mutant populations and generate digital transcriptomes for model species at key phylogenetic nodes. The proposed work will generate scenarios to explain how land plant form evolved and perhaps more importantly, how it could change in the future.
Max ERC Funding
2 230 732 €
Duration
Start date: 2009-07-01, End date: 2015-06-30
Project acronym GasPlaNt
Project Gas sensing in plants:Oxygen- and nitric oxide-regulated chromatin modification via a targeted protein degradation mechanism
Researcher (PI) Daniel James GIBBS
Host Institution (HI) THE UNIVERSITY OF BIRMINGHAM
Call Details Starting Grant (StG), LS3, ERC-2016-STG
Summary Oxygen (O2) and nitric oxide (NO) are gases that function as key developmental and stress-associated signals in plants. Investigating the molecular basis of their perception has the potential to identify new targets for crop improvement. In previous ground breaking work I showed that the direct transcriptional response to O2/NO is mediated by controlled degradation of specialised ‘gas-sensing’ transcription factors. We have now linked this degradation mechanism to a new functional class of ‘sensor’, a chromatin modifying protein that regulates the epigenetic silencing of genes. Here we will investigate the hypothesis that this protein acts as a previously undiscovered link between O2/NO and chromatin dynamics, and that plants have evolved a unique system for transducing gaseous signals into rapid transcriptional responses, and longer term epigenetic changes, through targeting different types of protein to the same degradation pathway.
Using multidisciplinary genetic, biochemical and omics approaches we will investigate the molecular basis of this novel gas perception system, which appears to be a plant-specific innovation. We will identify its global gene targets (the ‘gas-responsive epigenome’), and uncover its growth and stress-associated functions in Arabidopsis and barley. We will also investigate how manipulating this pathway using genome editing and synthetic biology techniques alters plant performance, focusing on traits of agronomic significance. This ambitious and timely research will take our knowledge of O2/NO-signaling and the control of chromatin dynamics beyond the current state of the art by offering insight into a completely novel signaling mechanism operating at the interface of gas-perception, protein degradation, and epigenetics. GasPlaNt will therefore provide a step-change in our understanding of how plants synchronise their gene expression in response to signals to optimise growth and development within a dynamic environment.
Summary
Oxygen (O2) and nitric oxide (NO) are gases that function as key developmental and stress-associated signals in plants. Investigating the molecular basis of their perception has the potential to identify new targets for crop improvement. In previous ground breaking work I showed that the direct transcriptional response to O2/NO is mediated by controlled degradation of specialised ‘gas-sensing’ transcription factors. We have now linked this degradation mechanism to a new functional class of ‘sensor’, a chromatin modifying protein that regulates the epigenetic silencing of genes. Here we will investigate the hypothesis that this protein acts as a previously undiscovered link between O2/NO and chromatin dynamics, and that plants have evolved a unique system for transducing gaseous signals into rapid transcriptional responses, and longer term epigenetic changes, through targeting different types of protein to the same degradation pathway.
Using multidisciplinary genetic, biochemical and omics approaches we will investigate the molecular basis of this novel gas perception system, which appears to be a plant-specific innovation. We will identify its global gene targets (the ‘gas-responsive epigenome’), and uncover its growth and stress-associated functions in Arabidopsis and barley. We will also investigate how manipulating this pathway using genome editing and synthetic biology techniques alters plant performance, focusing on traits of agronomic significance. This ambitious and timely research will take our knowledge of O2/NO-signaling and the control of chromatin dynamics beyond the current state of the art by offering insight into a completely novel signaling mechanism operating at the interface of gas-perception, protein degradation, and epigenetics. GasPlaNt will therefore provide a step-change in our understanding of how plants synchronise their gene expression in response to signals to optimise growth and development within a dynamic environment.
Max ERC Funding
1 495 341 €
Duration
Start date: 2017-03-01, End date: 2022-02-28
Project acronym HSCnicheIVM
Project In vivo imaging of haematopoietic stem cells in their natural niches to uncover cellular and molecular dynamics regulating self-renewal
Researcher (PI) Cristina Lo Celso
Host Institution (HI) IMPERIAL COLLEGE OF SCIENCE TECHNOLOGY AND MEDICINE
Call Details Starting Grant (StG), LS3, ERC-2013-StG
Summary Haematopoietic stem cells (HSC) reside in the bone marrow, from where they maintain immune cells, erythrocytes and platelets. To function correctly, they depend on their localisation within highly specialised niches, where cell-cell and -matrix interactions as well as medium- and long-range molecular signals are integrated to instruct them to either remain quiescent, or to generate progeny that will maintain both the stem cell pool and the differentiated lineages. Studies based on HSC transplantation assays have identified several signalling pathways and bone marrow cell types as regulators of HSC function; however the full picture of the cellular and molecular components of the HSC niche remains elusive because of lack of direct observation over time. HSC subpopulations have been identified based on their proliferative behaviour and it is likely that either migration between different microenvironments or transient modifications of the niche structure mediate changes in HSC fate in response to perturbations such as infection or leukaemia development.
I pioneered the combination of confocal and two-photon microscopy to visualise single HSC and their progeny within the bone marrow of live mice and here I propose to combine advanced microscopy techniques with multi-colour genetic lineage marking and highly sensitive expression profiling to track HSC and their clonal progeny in vivo in real time and to study the cellular and molecular composition of their niches during steady state and when responding to infection and leukaemia development. This work will uncover whether functionally distinct HSC subpopulations reside in anatomically distinct niches or rather all HSC niches are in principle equivalent, but change over time to mediate changes in HSC fate balance. The results obtained will provide a comprehensive picture of HSC niche dynamics, which will be critical for the development of regenerative medicine approaches based on in vivo or ex vivo expansion of HSC.
Summary
Haematopoietic stem cells (HSC) reside in the bone marrow, from where they maintain immune cells, erythrocytes and platelets. To function correctly, they depend on their localisation within highly specialised niches, where cell-cell and -matrix interactions as well as medium- and long-range molecular signals are integrated to instruct them to either remain quiescent, or to generate progeny that will maintain both the stem cell pool and the differentiated lineages. Studies based on HSC transplantation assays have identified several signalling pathways and bone marrow cell types as regulators of HSC function; however the full picture of the cellular and molecular components of the HSC niche remains elusive because of lack of direct observation over time. HSC subpopulations have been identified based on their proliferative behaviour and it is likely that either migration between different microenvironments or transient modifications of the niche structure mediate changes in HSC fate in response to perturbations such as infection or leukaemia development.
I pioneered the combination of confocal and two-photon microscopy to visualise single HSC and their progeny within the bone marrow of live mice and here I propose to combine advanced microscopy techniques with multi-colour genetic lineage marking and highly sensitive expression profiling to track HSC and their clonal progeny in vivo in real time and to study the cellular and molecular composition of their niches during steady state and when responding to infection and leukaemia development. This work will uncover whether functionally distinct HSC subpopulations reside in anatomically distinct niches or rather all HSC niches are in principle equivalent, but change over time to mediate changes in HSC fate balance. The results obtained will provide a comprehensive picture of HSC niche dynamics, which will be critical for the development of regenerative medicine approaches based on in vivo or ex vivo expansion of HSC.
Max ERC Funding
1 699 724 €
Duration
Start date: 2013-12-01, End date: 2018-11-30
Project acronym IniReg
Project Mechanisms of Regeneration Initiation
Researcher (PI) Kerstin BARTSCHERER
Host Institution (HI) KONINKLIJKE NEDERLANDSE AKADEMIE VAN WETENSCHAPPEN - KNAW
Call Details Starting Grant (StG), LS3, ERC-2016-STG
Summary Injury poses a key threat to all multicellular organisms. However, while some animals can fully restore lost body parts, others can only prevent further damage by mere wound healing. Which molecular mechanisms determine whether regeneration is induced or not is an unsettled fundamental question. I will use whole body regeneration, one of the most fascinating biological processes, as an experimental paradigm to identify the mechanisms of regeneration initiation. As a model organism I will employ planarians, flatworms with extraordinary plasticity that regenerate every piece of their body within a few days. I will mechanistically dissect how these animals rapidly induce an efficient regeneration program in response to tissue loss and define the key switches that determine whether a wound regenerates. Combining the astonishing regenerative abilities of planarians with new technologies I will first comprehensively describe the molecular changes occurring during the amputation response. Second, with a powerful novel assay developed in my lab - dormant fragments - that allows for the first time the separation of wounding from tissue loss in a single planarian, I will analyze the dynamics of the earliest regenerative events. Third, I will functionally characterize the regeneration-initiating signals and their target pathways combining in vivo RNAi and phenotypic assays. Fourth, with a regeneration-deficient planarian species, I will test whether the identified key regulators act as network nodes that can be utilized to rescue regeneration. Importantly, using vertebrate paradigms, such as the regenerating zebrafish fin, I will investigate conserved roles of these network nodes and validate general principles of regeneration initiation. This project will not only uncover conserved mechanisms of regeneration initiation but will also identify the switches that must be levered to induce regeneration in non-regenerating animals.
Summary
Injury poses a key threat to all multicellular organisms. However, while some animals can fully restore lost body parts, others can only prevent further damage by mere wound healing. Which molecular mechanisms determine whether regeneration is induced or not is an unsettled fundamental question. I will use whole body regeneration, one of the most fascinating biological processes, as an experimental paradigm to identify the mechanisms of regeneration initiation. As a model organism I will employ planarians, flatworms with extraordinary plasticity that regenerate every piece of their body within a few days. I will mechanistically dissect how these animals rapidly induce an efficient regeneration program in response to tissue loss and define the key switches that determine whether a wound regenerates. Combining the astonishing regenerative abilities of planarians with new technologies I will first comprehensively describe the molecular changes occurring during the amputation response. Second, with a powerful novel assay developed in my lab - dormant fragments - that allows for the first time the separation of wounding from tissue loss in a single planarian, I will analyze the dynamics of the earliest regenerative events. Third, I will functionally characterize the regeneration-initiating signals and their target pathways combining in vivo RNAi and phenotypic assays. Fourth, with a regeneration-deficient planarian species, I will test whether the identified key regulators act as network nodes that can be utilized to rescue regeneration. Importantly, using vertebrate paradigms, such as the regenerating zebrafish fin, I will investigate conserved roles of these network nodes and validate general principles of regeneration initiation. This project will not only uncover conserved mechanisms of regeneration initiation but will also identify the switches that must be levered to induce regeneration in non-regenerating animals.
Max ERC Funding
1 500 000 €
Duration
Start date: 2017-04-01, End date: 2022-03-31
Project acronym LogNeuroDev
Project The molecular and cellular logic of vertebrate neural development
Researcher (PI) James BRISCOE
Host Institution (HI) THE FRANCIS CRICK INSTITUTE LIMITED
Call Details Advanced Grant (AdG), LS3, ERC-2016-ADG
Summary A central problem in biology and key to realising the potential of regenerative medicine is understanding the mechanisms that produce and organize cells in the complex tissues of an embryo. In broad terms, initially uncommitted progenitors acquire their fate in response to signals that control transcriptional programmes. These programmes drive cells through spatial and temporal successions of states that gradually refine cell identity. How these states are established and cell fate decisions implemented is poorly understood. To address this we use an experimentally tractable system – the formation of defined populations of progenitors in the vertebrate spinal cord. We take an interdisciplinary approach that combines our in vivo expertise with three recent advances in our group. First, we have developed in vitro differentiation systems and microfluidic devices that use embryonic stem cells to recapitulate development processes. Second, we have embraced new technologies that provide unprecedented ability to manipulate and assay single cells. Finally, we have established interdisciplinary collaborations to develop computational tools and construct data driven mathematical models. Using these approaches, alongside established embryological methods, we will establish a platform for manipulating and analysing mechanisms by which the multipotent progenitors that form the spinal cord acquire specific identities. We will identify the rules by which cells make decisions and we will define the design logic and network architectures that lead to distinct cell fate choices. The ability to: (i) follow the trajectory of a cell as it transitions to a specific neuronal subtype in vivo; (ii) manipulate the process in vitro and in vivo; and (iii) model it in silico, offers a unique system for understanding organogenesis. Together these approaches will provide the knowledge and technical foundations for rational, predictive tissue engineering of the spinal cord.
Summary
A central problem in biology and key to realising the potential of regenerative medicine is understanding the mechanisms that produce and organize cells in the complex tissues of an embryo. In broad terms, initially uncommitted progenitors acquire their fate in response to signals that control transcriptional programmes. These programmes drive cells through spatial and temporal successions of states that gradually refine cell identity. How these states are established and cell fate decisions implemented is poorly understood. To address this we use an experimentally tractable system – the formation of defined populations of progenitors in the vertebrate spinal cord. We take an interdisciplinary approach that combines our in vivo expertise with three recent advances in our group. First, we have developed in vitro differentiation systems and microfluidic devices that use embryonic stem cells to recapitulate development processes. Second, we have embraced new technologies that provide unprecedented ability to manipulate and assay single cells. Finally, we have established interdisciplinary collaborations to develop computational tools and construct data driven mathematical models. Using these approaches, alongside established embryological methods, we will establish a platform for manipulating and analysing mechanisms by which the multipotent progenitors that form the spinal cord acquire specific identities. We will identify the rules by which cells make decisions and we will define the design logic and network architectures that lead to distinct cell fate choices. The ability to: (i) follow the trajectory of a cell as it transitions to a specific neuronal subtype in vivo; (ii) manipulate the process in vitro and in vivo; and (iii) model it in silico, offers a unique system for understanding organogenesis. Together these approaches will provide the knowledge and technical foundations for rational, predictive tissue engineering of the spinal cord.
Max ERC Funding
2 357 000 €
Duration
Start date: 2018-01-01, End date: 2022-12-31
Project acronym MINICELL
Project Building minimal cells to understand active cell shape control
Researcher (PI) Gijsberta Koenderink
Host Institution (HI) STICHTING NEDERLANDSE WETENSCHAPPELIJK ONDERZOEK INSTITUTEN
Call Details Starting Grant (StG), LS3, ERC-2013-StG
Summary Understanding how cells control their shape is an important scientific goal, since cells in our body constantly need to undergo shape changes to perform vital tasks such as growth and division. Conversely, abnormal cell shape changes contribute to life-threatening diseases such as cancer and developmental disorders. I propose to resolve the physical basis of active cell shape control by studying minimal cells built from purified cellular components. The main determinant of cell shape in animals is the actin cortex beneath the cell membrane, which contains molecular motors that actively generate forces. There is growing evidence that cells tightly balance these active forces with passive forces arising from cortex-membrane adhesion and elasticity. However, it is unclear how these forces are generated and controlled on the molecular level given the enormous complexity of cells. To circumvent this complexity, we will reconstitute cell-free actin networks and couple them to model biomembranes with the essential cellular linker protein septin. Using various advanced microscopy techniques, we will study (1) how active cortical networks and lipid bilayers influence each other’s spatial organization; (2) how active cortical networks control membrane shape; and (3) how spatial gradients in cortex contractility can cause cell shape polarization. My long-term ambition is to bridge the gap between the physical properties of cell-free model systems and biological functions in living cells. Thanks to recent breakthroughs in our understanding of the biophysical properties of contractile actin networks, we can now build more relevant cell-free model systems that can mimic active cell shape changes. To test the biological relevance of our findings, we will confront our results with live cell observations in fly embryos, together with a developmental biology group. Ultimately, the model cells developed here will enable a wide range of further studies of cellular (mal)functions.
Summary
Understanding how cells control their shape is an important scientific goal, since cells in our body constantly need to undergo shape changes to perform vital tasks such as growth and division. Conversely, abnormal cell shape changes contribute to life-threatening diseases such as cancer and developmental disorders. I propose to resolve the physical basis of active cell shape control by studying minimal cells built from purified cellular components. The main determinant of cell shape in animals is the actin cortex beneath the cell membrane, which contains molecular motors that actively generate forces. There is growing evidence that cells tightly balance these active forces with passive forces arising from cortex-membrane adhesion and elasticity. However, it is unclear how these forces are generated and controlled on the molecular level given the enormous complexity of cells. To circumvent this complexity, we will reconstitute cell-free actin networks and couple them to model biomembranes with the essential cellular linker protein septin. Using various advanced microscopy techniques, we will study (1) how active cortical networks and lipid bilayers influence each other’s spatial organization; (2) how active cortical networks control membrane shape; and (3) how spatial gradients in cortex contractility can cause cell shape polarization. My long-term ambition is to bridge the gap between the physical properties of cell-free model systems and biological functions in living cells. Thanks to recent breakthroughs in our understanding of the biophysical properties of contractile actin networks, we can now build more relevant cell-free model systems that can mimic active cell shape changes. To test the biological relevance of our findings, we will confront our results with live cell observations in fly embryos, together with a developmental biology group. Ultimately, the model cells developed here will enable a wide range of further studies of cellular (mal)functions.
Max ERC Funding
1 448 000 €
Duration
Start date: 2013-12-01, End date: 2018-11-30
Project acronym NeuroMT
Project Building the Neuronal Microtubule Cytoskeleton
Researcher (PI) Casper Cassander Hoogenraad
Host Institution (HI) UNIVERSITEIT UTRECHT
Call Details Consolidator Grant (CoG), LS3, ERC-2013-CoG
Summary Microtubules (MTs) are one of the major cytoskeletal components of the cell, essential for many fundamental cellular and developmental processes, such as cell division, motility and polarity. In large and highly polarized cells like neurons, MTs have been regarded as essential structures for stable neuronal morphology and serve as tracks for long-distance transport; however, fundamental new insights into the role of neural MTs have emerged. New findings demonstrate that the MT cytoskeleton plays an active role during different phases of neuronal development: MTs determine axon formation, control polarized cargo trafficking, and regulate the dynamics of dendritic spines, the major sites of excitatory synaptic input. Failures in MT function have been linked to various neurological and neurodegenerative diseases and recent studies highlight MTs as a potential target for therapeutic intervention.
How neuronal MTs are formed and stabilized during neuronal polarity and differentiation is largely unknown, and whether this requires the centrosome is under debate. The overall aim of this proposal is to investigate basic mechanisms responsible for organizing the microtubule cytoskeleton during neuronal development. Here, we will take a multidisciplinary approach and combine biochemistry, neurobiology, molecular engineering, and advanced microscopy to study the role of MTs at three stages of neuronal differentiation. We propose to determine: i) the role of (non-)centrosomal MT nucleation during early development, ii) the mechanism by which dendrites organize MTs into anti-parallel arrays, iii) the relation between MTs spine entry and cargo transport in mature neurons. We anticipate that these studies will uncover how the MT cytoskeleton is built and organized at different phases of neuronal development, which will be relevant for understanding polarized transport, synaptic processes and associated neurodegenerative disorders.
Summary
Microtubules (MTs) are one of the major cytoskeletal components of the cell, essential for many fundamental cellular and developmental processes, such as cell division, motility and polarity. In large and highly polarized cells like neurons, MTs have been regarded as essential structures for stable neuronal morphology and serve as tracks for long-distance transport; however, fundamental new insights into the role of neural MTs have emerged. New findings demonstrate that the MT cytoskeleton plays an active role during different phases of neuronal development: MTs determine axon formation, control polarized cargo trafficking, and regulate the dynamics of dendritic spines, the major sites of excitatory synaptic input. Failures in MT function have been linked to various neurological and neurodegenerative diseases and recent studies highlight MTs as a potential target for therapeutic intervention.
How neuronal MTs are formed and stabilized during neuronal polarity and differentiation is largely unknown, and whether this requires the centrosome is under debate. The overall aim of this proposal is to investigate basic mechanisms responsible for organizing the microtubule cytoskeleton during neuronal development. Here, we will take a multidisciplinary approach and combine biochemistry, neurobiology, molecular engineering, and advanced microscopy to study the role of MTs at three stages of neuronal differentiation. We propose to determine: i) the role of (non-)centrosomal MT nucleation during early development, ii) the mechanism by which dendrites organize MTs into anti-parallel arrays, iii) the relation between MTs spine entry and cargo transport in mature neurons. We anticipate that these studies will uncover how the MT cytoskeleton is built and organized at different phases of neuronal development, which will be relevant for understanding polarized transport, synaptic processes and associated neurodegenerative disorders.
Max ERC Funding
2 000 000 €
Duration
Start date: 2014-11-01, End date: 2019-10-31
Project acronym PHYSBIOHSC
Project Understanding the physical biology of adult blood stem cells
Researcher (PI) David KENT
Host Institution (HI) THE CHANCELLOR MASTERS AND SCHOLARS OF THE UNIVERSITY OF CAMBRIDGE
Call Details Starting Grant (StG), LS3, ERC-2016-STG
Summary The discovery of functional heterogeneity in normal and malignant stem cells has shifted our understanding of how single cells are subverted to drive cancer. To design therapies for diseases of stem cell origin and to better provide cell populations for clinical applications, it is critical to understand this diversity at the single cell level. This proposal focuses on understanding the complex biology of normal and malignant stem cells and the impact of individual mutations on clonal evolution by studying the physical and quantitative aspects of single blood stem cells.
This proposal aims to study single blood stem cell biomechanics and clonal evolution by leveraging new inter-disciplinary technologies and approaches and applying them to functionally defined mouse and human blood stem cell populations. It will combine in vitro and in vivo biological assays with mathematical modelling and microfluidic technology in an iterative manner across both human and mouse stem cell populations.
Summary
The discovery of functional heterogeneity in normal and malignant stem cells has shifted our understanding of how single cells are subverted to drive cancer. To design therapies for diseases of stem cell origin and to better provide cell populations for clinical applications, it is critical to understand this diversity at the single cell level. This proposal focuses on understanding the complex biology of normal and malignant stem cells and the impact of individual mutations on clonal evolution by studying the physical and quantitative aspects of single blood stem cells.
This proposal aims to study single blood stem cell biomechanics and clonal evolution by leveraging new inter-disciplinary technologies and approaches and applying them to functionally defined mouse and human blood stem cell populations. It will combine in vitro and in vivo biological assays with mathematical modelling and microfluidic technology in an iterative manner across both human and mouse stem cell populations.
Max ERC Funding
1 500 000 €
Duration
Start date: 2017-05-01, End date: 2022-04-30
Project acronym PolarizedTransport
Project Sorting out Polarized Transport in Neurons: Motor Protein Selectivity and Cooperativity
Researcher (PI) Lukas Christiaan Kapitein
Host Institution (HI) UNIVERSITEIT UTRECHT
Call Details Starting Grant (StG), LS3, ERC-2013-StG
Summary Neurons are the building blocks of the brain. The ability of neurons to receive, process and transmit information depends on their polarized organization into axons and dendrites. To build such a highly polarized cell, cellular components synthesized in the cell body are differentially transported to either axons or dendrites. Polarized transport is driven by three families of cytoskeletal motor proteins, which can walk in different directions over the actin or microtubule cytoskeleton. Many subfamilies of motor proteins exist, but how each of these motor proteins contributes to selective cargo delivery is unknown.
I have recently developed an approach to probe specific motor activity inside cells, which revealed that many microtubule-based motors selectively target axons. However, the molecular mechanisms behind this remarkable selectivity are unknown. In addition, it is well-established that most cargos are transported by a combination of different motors, but how the activity of different types of motors on the same cargo is integrated has remained unclear.
The aim of this proposal is to understand how motor proteins navigate the neuronal cytoskeleton. We will take a multidisciplinary approach and combine neurobiology, molecular engineering, advanced microscopy, and mathematical modelling to study the origin of motor selectivity as well as the collective activity of dissimilar motor teams. We will employ and expand our unique methodology to: 1) Study how the spatial organization and post-translational modifications of the microtubule cytoskeleton facilitate selective transport. 2) Perform well-controlled intracellular multi-motor assays to understand the functional interplay between different types of motors.
Successful achievement of these objectives will uncover key mechanisms of polarized transport, which will be relevant for understanding transport-associated neurodegenerative diseases.
Summary
Neurons are the building blocks of the brain. The ability of neurons to receive, process and transmit information depends on their polarized organization into axons and dendrites. To build such a highly polarized cell, cellular components synthesized in the cell body are differentially transported to either axons or dendrites. Polarized transport is driven by three families of cytoskeletal motor proteins, which can walk in different directions over the actin or microtubule cytoskeleton. Many subfamilies of motor proteins exist, but how each of these motor proteins contributes to selective cargo delivery is unknown.
I have recently developed an approach to probe specific motor activity inside cells, which revealed that many microtubule-based motors selectively target axons. However, the molecular mechanisms behind this remarkable selectivity are unknown. In addition, it is well-established that most cargos are transported by a combination of different motors, but how the activity of different types of motors on the same cargo is integrated has remained unclear.
The aim of this proposal is to understand how motor proteins navigate the neuronal cytoskeleton. We will take a multidisciplinary approach and combine neurobiology, molecular engineering, advanced microscopy, and mathematical modelling to study the origin of motor selectivity as well as the collective activity of dissimilar motor teams. We will employ and expand our unique methodology to: 1) Study how the spatial organization and post-translational modifications of the microtubule cytoskeleton facilitate selective transport. 2) Perform well-controlled intracellular multi-motor assays to understand the functional interplay between different types of motors.
Successful achievement of these objectives will uncover key mechanisms of polarized transport, which will be relevant for understanding transport-associated neurodegenerative diseases.
Max ERC Funding
1 500 000 €
Duration
Start date: 2014-02-01, End date: 2019-01-31
Project acronym RepTime
Project Molecular control of DNA replication timing in mammalian cells
Researcher (PI) Sara Cristiana Barbara BUONOMO
Host Institution (HI) THE UNIVERSITY OF EDINBURGH
Call Details Consolidator Grant (CoG), LS3, ERC-2016-COG
Summary DNA replication is an essential process ensuring the transmission of genetic information and is highly regulated. Specifically, the DNA replication-timing program ensures that the sites of initiation of DNA replication, termed origins, are not all activated simultaneously but follow a cell-type specific schedule. This pathway is conserved throughout eukaryotic evolution, however its molecular control and biological role are not fully understood. In this proposal I aim to understand key aspects of replication-timing program by employing a combination of advanced mouse genetics, genomics, cell biology and proteomics. Currently one of the major limitations in the mammalian DNA replication field is the elusive identity of origins. I aim to comprehensively map origins in a variety of mouse cells/tissues and relate the regulation of origin firing to the control of gene expression and three-dimensional nuclear architecture. I have discovered that Rif1 controls replication timing and links it to nuclear three-dimensional organization. I have also revealed the existence of a novel Rif1-independent pathway that controls the timing of a significant fraction of the late-replicating genome, identified by constitutive association with a key nuclear architecture component, Lamin B1. Here, I propose complementary approaches to understand the molecular mechanism by which Rif1 coordinates replication timing and nuclear organization as well as the molecular underpinnings of the novel pathway instructing late-replication in Lamin B1-associated regions. Finally, my goal is to understand the in vivo biological role of the replication-timing program. Our preliminary data identify mammalian X inactivation as a process where replication timing may play a fundamental part. My ultimate objective is to contribute to the realization of a comprehensive understanding of nuclear function, integrating the co-regulation of DNA replication with gene expression, epigenetic inheritance and DNA repair.
Summary
DNA replication is an essential process ensuring the transmission of genetic information and is highly regulated. Specifically, the DNA replication-timing program ensures that the sites of initiation of DNA replication, termed origins, are not all activated simultaneously but follow a cell-type specific schedule. This pathway is conserved throughout eukaryotic evolution, however its molecular control and biological role are not fully understood. In this proposal I aim to understand key aspects of replication-timing program by employing a combination of advanced mouse genetics, genomics, cell biology and proteomics. Currently one of the major limitations in the mammalian DNA replication field is the elusive identity of origins. I aim to comprehensively map origins in a variety of mouse cells/tissues and relate the regulation of origin firing to the control of gene expression and three-dimensional nuclear architecture. I have discovered that Rif1 controls replication timing and links it to nuclear three-dimensional organization. I have also revealed the existence of a novel Rif1-independent pathway that controls the timing of a significant fraction of the late-replicating genome, identified by constitutive association with a key nuclear architecture component, Lamin B1. Here, I propose complementary approaches to understand the molecular mechanism by which Rif1 coordinates replication timing and nuclear organization as well as the molecular underpinnings of the novel pathway instructing late-replication in Lamin B1-associated regions. Finally, my goal is to understand the in vivo biological role of the replication-timing program. Our preliminary data identify mammalian X inactivation as a process where replication timing may play a fundamental part. My ultimate objective is to contribute to the realization of a comprehensive understanding of nuclear function, integrating the co-regulation of DNA replication with gene expression, epigenetic inheritance and DNA repair.
Max ERC Funding
1 999 785 €
Duration
Start date: 2017-10-01, End date: 2022-09-30
