Investigation of the function of the ER/mitochondria contact sites in cell physiology and disease
Lead Research Organisation:
University of Cambridge
Department Name: UNLISTED
Abstract
Mitochondria are highly dynamic organelles essential for the survival of the cell. Mitochondrial dynamics has been linked to different physiological functions, and contacts with other organelles are required for specific metabolic activity and mitochondrial behaviour. Indeed, endoplasmic reticulum and mitochondria work together and their closed juxtaposition is considered as a signalling platform for metabolite flux. Alteration of these contacts has been associated with defects in cellular homeostasis and with human disease. The goal of our group is to characterise the architecture and understand the associated functions of these inter-organelle contacts. Using biochemical, molecular and cell biological approaches, we want to investigate the function of the mitochondria/endoplasmic reticulum contact sites in different cell physiology processes including cell death, lipid and calcium fluxes and cell migration. This work will lead us to obtaining new insights into the importance of mitochondria and mitochondria/ER contacts in cellular signalling pathways, and to better understanding the pathomechanisms underpinning human diseases associated with defects in their regulation and dynamics.
Technical Summary
Mitochondria are complex organelles that provide essentially all of the energy, mainly through ATP synthesis, required for normal physiological function in nearly all cells of the body. They also play a central role in a number of cell signaling pathways, including cell death and regulation of the intracellular calcium homeostasis. Mitochondria are highly dynamic organelles that can adapt their network in response to specific cellular needs. Indeed, they are able to constantly fuse and divide, and these events are intimately related to their metabolic functions and cellular stress signals. Impairment of mitochondrial physiology is linked to human diseases including neurodegenerative and metabolic disorders such as Parkinson’s disease and diabetes, and also in the progression of certain forms of cancer. It has become clear recently that mitochondria do not function in isolation, but establish direct contact and communication with other cellular organelles to exchange metabolites and signals converging on, or streaming from, mitochondria. For instance, mitochondria are physically coupled to the endoplasmic reticulum through contact sites that act as a signaling platform. These are called mitochondria-associated membranes (MAM), and are essential for a number of processes, including intracellular calcium homeostasis, lipid exchange, mitochondrial dynamics and motility, ROS production and apoptosis. Studies performed in the last decade have identified several proteins which directly tether the two organelles in MAMs and proteins enriched in these contact sites that play highly specialised homeostatic or execution roles. However, the biogenesis, regulation and precise molecular mechanisms associated with these signaling platforms are still poorly understood. The goal of our lab is to understand the physiological importance of these inter-organelle contacts in specific cellular contexts, including metabolite flux during steady state, and in response to specific triggers, particularly during cell death or cell migration. Using molecular biological and biochemical approaches we will focus on the characterisation of proteins localised at the interface of the mito/ER contacts sites, their potential role in the stability of the MAM structures, and their functions in lipid and calcium fluxes during the execution of the cell death programme. We will couple these techniques with the state-of-the-art advanced microscopic analysis, including spinning disc and scanning confocal systems, super-resolution microscopy, and electron microscopy. To unravel the function of mito/ER contacts sites on cell physiology, live cell imaging will be performed to study organelle movements during cell migration. The precise localisation of target proteins will be studied by the emergent super-resolution microscopy techniques. Finally, we are also interested in understanding the relevance of mitochondrial dynamics and MAMs in the context of mitochondrial disease. We will study the mitochondrial biology, in different cellular models derived from patients, to characterize how mitochondrial morphology and inter-organelle contacts are affected by different gene mutations, and how this contributes to disease progression.
People |
ORCID iD |
| Julien Prudent (Principal Investigator) |
Publications
Hathazi D
(2020)
Metabolic shift underlies recovery in reversible infantile respiratory chain deficiency.
in The EMBO journal
Van Den Ameele J
(2020)
Chronic pain is common in mitochondrial disease.
Reyes A
(2020)
RNase H1 Regulates Mitochondrial Transcription and Translation via the Degradation of 7S RNA
in Frontiers in Genetics
Wei W
(2020)
Nuclear-mitochondrial DNA segments resemble paternally inherited mitochondrial DNA in humans
in Nature Communications
Biner O
(2020)
Bottom-Up Construction of a Minimal System for Cellular Respiration and Energy Regeneration.
in ACS synthetic biology
Russell D
(2020)
Hydroxylated Rotenoids Selectively Inhibit the Proliferation of Prostate Cancer Cells
in Journal of Natural Products
Loreto A
(2020)
Mitochondrial impairment activates the Wallerian pathway through depletion of NMNAT2 leading to SARM1-dependent axon degeneration
in Neurobiology of Disease
Protasoni M
(2020)
Novel compound heterozygous pathogenic variants in nucleotide-binding protein like protein (NUBPL) cause leukoencephalopathy with multi-systemic involvement.
in Molecular genetics and metabolism
Murphy M
(2020)
How should we talk about metabolism?
in Nature Immunology
King M
(2020)
A Single Cysteine Residue in the Translocation Pathway of the Mitosomal ADP/ATP Carrier from Cryptosporidium parvum Confers a Broad Nucleotide Specificity
in International Journal of Molecular Sciences
Grba DN
(2020)
Mitochondrial complex I structure reveals ordered water molecules for catalysis and proton translocation.
in Nature structural & molecular biology
De Lazzari F
(2020)
Antioxidant Therapy in Parkinson's Disease: Insights from Drosophila melanogaster
in Antioxidants
Diederichs KA
(2020)
Structural insight into mitochondrial ß-barrel outer membrane protein biogenesis.
in Nature communications
F C Lopes A
(2020)
Mitochondrial metabolism and DNA methylation: a review of the interaction between two genomes.
in Clinical epigenetics
Powell C
(2020)
TRMT2B is responsible for both tRNA and rRNA m 5 U-methylation in human mitochondria
in RNA Biology
Walker JE
(2020)
Reply to Bernardi: The mitochondrial permeability transition pore and the ATP synthase.
in Proceedings of the National Academy of Sciences of the United States of America
Silva-Pinheiro P
(2020)
In vivo and in vitro mechanistic characterization of a clinically relevant Pol?A mutation
Bacman SR
(2020)
Manipulation of mitochondrial genes and mtDNA heteroplasmy.
in Methods in cell biology
Szibor M
(2020)
Respiratory chain signalling is essential for adaptive remodelling following cardiac ischaemia.
in Journal of cellular and molecular medicine
Szibor M
(2020)
Bioenergetic consequences from xenotopic expression of a tunicate AOX in mouse mitochondria: Switch from RET and ROS to FET
in Biochimica et Biophysica Acta (BBA) - Bioenergetics
Terriente-Felix A
(2020)
Drosophila phosphatidylinositol-4 kinase fwd promotes mitochondrial fission and can suppress Pink1/parkin phenotypes.
in PLoS genetics
Spikes T
(2020)
Structure of the dimeric ATP synthase from bovine mitochondria
in Proceedings of the National Academy of Sciences
Ruprecht J
(2020)
The SLC25 Mitochondrial Carrier Family: Structure and Mechanism.
Tábara LC
(2020)
The last wall of defense to prevent extreme and deleterious mitochondrial fusion.
in The EMBO journal
Lee J
(2020)
The STING pathway does not contribute to behavioural or mitochondrial phenotypes in Drosophila Pink1/parkin or mtDNA mutator models
in Scientific Reports
Zanette V
(2020)
Neurodevelopmental regression, severe generalized dystonia, and metabolic acidosis caused by POLR3A mutations.
in Neurology. Genetics
Robinson WE
(2020)
Understanding How the Rate of C-H Bond Cleavage Affects Formate Oxidation Catalysis by a Mo-Dependent Formate Dehydrogenase.
in Journal of the American Chemical Society
Protasoni M
(2020)
Respiratory supercomplexes act as a platform for complex III-mediated maturation of human mitochondrial complexes I and IV.
in The EMBO journal
Wei W
(2020)
Inheritance of mitochondrial DNA in humans: implications for rare and common diseases
in Journal of Internal Medicine
Spikes T
(2020)
Structure of the dimeric ATP synthase from bovine mitochondria
King MS
(2020)
Expression and Purification of Membrane Proteins in Saccharomyces cerevisiae.
in Methods in molecular biology (Clifton, N.J.)
Di Nottia M
(2020)
A homozygous MRPL24 mutation causes a complex movement disorder and affects the mitoribosome assembly
in Neurobiology of Disease
Baxter M
(2020)
Cardiac mitochondrial function depends on BUD23 mediated ribosome programming
in eLife
Van Den Ameele J
(2020)
Chronic pain is common in mitochondrial disease
in Neuromuscular Disorders
Nagashima S
(2020)
Golgi-derived PI ( 4 ) P-containing vesicles drive late steps of mitochondrial division
in Science
| Description | Daiichi Sankyo Foundation of Life Science fellowship programme |
| Amount | £40,125 (GBP) |
| Organisation | Daiichi Sankyo Company |
| Sector | Private |
| Country | Japan |
| Start | 08/2017 |
| End | 09/2019 |
| Description | Isaac Newton Trust / Wellcome Trust ISSF / University of Cambridge Joint Research |
| Amount | £85,000 (GBP) |
| Organisation | University of Edinburgh |
| Department | Institutional Strategic Support Fund |
| Sector | Charity/Non Profit |
| Country | United Kingdom |
| Start | 03/2018 |
| End | 03/2020 |
| Description | Postdoctoral Marie Sklodowska-Curie fellowship 2017 |
| Amount | £183,455 (GBP) |
| Funding ID | MITODYN |
| Organisation | Marie Sklodowska-Curie Actions |
| Sector | Charity/Non Profit |
| Country | Global |
| Start | 06/2017 |
| End | 07/2019 |
| Description | Ramon Areces Foundation postdoctoral fellowship |
| Amount | € 48,000 (EUR) |
| Organisation | Ramón Areces Foundation |
| Sector | Charity/Non Profit |
| Country | Spain |
| Start | 09/2019 |
| End | 10/2021 |
| Description | Characterization of TMEM63C |
| Organisation | University of Exeter |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | With Pr. Andrew Crosby and Dr. Emma Baple (University of Exeter, UK), we have identified and characterized TMEM63C as a new disease gene involved in hereditary Spastic Paraplegia. Co-lead the project, Scientific expertize, intellectual input, exchange of ideas, perform experiments. |
| Collaborator Contribution | Co-lead the project, Scientific expertize, intellectual input, exchange of ideas, perform experiments. |
| Impact | Manuscript just accepted for publication in Brain. |
| Start Year | 2019 |
| Description | Detection of membrane contact sites |
| Organisation | University of Cambridge |
| Department | Department of Clinical Neurosciences |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | Collaboration with Pr. Patrick Yu Wai Man (Clinical neurosciences, University of Cambridge) on the contribution of MERCs to optic neuropathies. Scientific expertize, intellectual input, exchange of ideas, perform experiments, sharing of reagents, host student |
| Collaborator Contribution | Initiate the project, Scientific expertize, intellectual input, exchange of ideas, perform experiments. |
| Impact | Publication: PMID: 34656549 |
| Start Year | 2017 |
| Description | Linking mTOR signalling, mitochondrial dynamics and cell survival |
| Organisation | McGill University |
| Country | Canada |
| Sector | Academic/University |
| PI Contribution | In this collaboration, I provided scientific expertise, intellectual input and performed experiments for our publication. |
| Collaborator Contribution | Collaborators designed the project, contributed to scientific expertize and intellectual input, allowed facility access and performed the experiments. |
| Impact | Publication: doi: 10.1016/j.molcel.2017.08.013. PMID: 28918902. |
| Start Year | 2014 |
| Description | Mitochondria and CTL killing capacities |
| Organisation | University of Cambridge |
| Department | Cambridge Institute for Medical Research (CIMR) |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | Scientific expertize, intellectual input, exchange of ideas, sharing of reagents, perform experiments |
| Collaborator Contribution | Our collaborator has established, developed and performed the project |
| Impact | Multi-disciplinary project from Immunology to cell biology and mitochondrial biology. This collaboration let to a scientific publication (PMID: 34648346). |
| Start Year | 2018 |
| Description | Mitochondrial calcium and cancer cell migration |
| Organisation | Claude Bernard University Lyon 1 (UCBL) |
| Country | France |
| Sector | Academic/University |
| PI Contribution | I contributed to design and start the project, scientific expertize and intelectual input. |
| Collaborator Contribution | They performed all the experiements and contributed to scientific expertize |
| Impact | Publications: doi: 10.1038/srep36570. PMID: 27827394 DOI: 10.5772/intechopen.74494 |
| Start Year | 2012 |
| Description | Mitochondrial dynamics and contact sites |
| Organisation | McGill University |
| Department | Montreal Neurological Institute and Hospital |
| Country | Canada |
| Sector | Hospitals |
| PI Contribution | Scientific expertize, intellectual input, exchange of ideas |
| Collaborator Contribution | Scientific expertize, intellectual input, exchange of ideas |
| Impact | Publication: doi: 10.1016/j.ceb.2017.03.007. PMID: 28391089 |
| Start Year | 2013 |
| Description | Mitofusins and metabolism |
| Organisation | University of Cambridge |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | In collaboration with Pr. David Savage from the Metabolic Research Laboratories, Wellcome Trust-Medical Research Council Institute of Metabolic Science, University of Cambridge, UK, and Pr. R Semple (University of Edinburgh, UK) we explore how mutant/loss of Mitofusins and mitochondrial fusion affect adipogenesis. In my lab, we investigate how mutation or the loss in Mfns control different mitochondrial functions, including bioenergetics, morphology and membrane contact sites with other organelles, using a specific mice model with Mfn2 mutation or in vitro models, generated by the lab of Pr. David Savage. |
| Collaborator Contribution | Pr D.Savage and R. Semple has generated and characterised different models of mitochondrial dysfunction, including Mfn2 mutant KI mice or different in vitro cell models, top investigate how Mfns and mitochondrial fusion control Adipocyte biology and metabolism. They are leading the collaboration. |
| Impact | This a multi-disciplenary project combining in vivo, metabolism adipocytes biology, mitochondria, bioenergetics and microscopy. This collaboration let to two publications, one published in elife and one in Biorxiv. |
| Start Year | 2020 |
| Description | Access Students from Cambridge Regional College |
| Form Of Engagement Activity | Participation in an open day or visit at my research institution |
| Part Of Official Scheme? | No |
| Geographic Reach | Regional |
| Primary Audience | Schools |
| Results and Impact | 11 Access students from Cambridge Regional College visited the MBU for a "Biology Masterclass", a visit to the fly laboratory and a "meet the scientists" session, where hands-on "festival" activities were available. |
| Year(s) Of Engagement Activity | 2022 |
| Description | Big Biology Day |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | Regional |
| Primary Audience | Public/other audiences |
| Results and Impact | Members of the MRC MBU participated in the Big Biology Day 2017 at Hills Road Sixth Form College. Scientists explained the Unit's research with hands-on activities. These included experiments using Drosophila melanogaster - with fluorescent markings and temperature sensitivity, our "Find the mutant fly" game, colouring books and posters. Feedback: Interesting, fun, enjoyable. 2019: Members of the MBU took Mito-gami, "Destroying Mutant Mitochondrial DNA", DNA sequencing puzzles and colouring/activity books. There was much conversation about the science and career options. |
| Year(s) Of Engagement Activity | 2017,2018,2019 |
| URL | http://www.hillsroad.ac.uk/college-life/events/2017/10/14/default-calendar/big-biology-day |
| Description | Big Biology Day 2022, 2023, 2024 |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | National |
| Primary Audience | Schools |
| Results and Impact | Hands on, interactive activities showcasing the MBU's research on mitochondria, careers advice. Activities will include our MITOTrumps card game, demonstrations using fruit flies and mitochondrial pinball. |
| Year(s) Of Engagement Activity | 2022,2024 |
| URL | https://www.mrc-mbu.cam.ac.uk/news/mrc-mbu-big-biology-day-2022 |
| Description | Dissemination of scientific achievements via the Unit's website and social media |
| Form Of Engagement Activity | Engagement focused website, blog or social media channel |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Public/other audiences |
| Results and Impact | https://www.facebook.com/mrc-mbu Twitter - @LabPrudent and @MRC_MBU https://www.mrc-mbu.cam.ac.uk/research-groups/prudent-group https://www.mrc-mbu.cam.ac.uk |
| Year(s) Of Engagement Activity | 2017,2018,2019,2020,2021,2022 |
| URL | https://www.mrc-mbu.cam.ac.uk/research-groups/prudent-group |
| Description | The Big Bang Fair London - Newham |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | Regional |
| Primary Audience | Schools |
| Results and Impact | Demonstration of genetic influence on body characteristics in Drosophila, "Find My Mutant Fly" game, posters explaining the science, movies. |
| Year(s) Of Engagement Activity | 2017 |
| URL | http://nearme.thebigbangfair.co.uk/london/ |
| Description | University of Cambridge Science Festival |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Public/other audiences |
| Results and Impact | Up close and personal with fluorescent fruit flies. Members of the MBU showcased the Unit's research using fluorescent fruit flies - examined under a microscope. Temperature sensitive fruit flies were also used to demonstrate the effects of Parkinson's disease. Other activities included movies, posters, a game of elimination and an MRC Cambridge colouring book. 2019: Further showcasing of the Unit's research through the use of hands-on games and activities, discussions and explanations. |
| Year(s) Of Engagement Activity | 2017,2018,2019 |
| Description | Website and social media |
| Form Of Engagement Activity | Engagement focused website, blog or social media channel |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Public/other audiences |
| Results and Impact | My research is promoted on the MBU's website and via social media channels, such as Facebook and Twitter. This leads to various forms of engagement - increased awareness, requests for further information, potential collaborations etc. |
| Year(s) Of Engagement Activity | 2022,2023,2024,2025 |
| URL | https://www.mrc-mbu.cam.ac.uk/research-groups/prudent-group |
| Description | YSGOL John Bright Comprehensive School: Virtual Biology Masterclass |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | Regional |
| Primary Audience | Schools |
| Results and Impact | A level Biology students at YSGOL John Bright Comprehensive School, in Llandudno (North Wales) received a "Biology Masterclass" from Jordan Morris (a PhD student in my research group), who discussed with them the many roles of mitochondria in the cell, and how his work on the fundamental principles of mitochondrial biology has implications for the treatment of cancer. Feedback from tutor: The students really engaged with Jordan's excellent presentation; it was well linked to their A level studies, allowing them to engage with concepts they have begun to understand, whilst challenging them to think about the bigger picture of how their studies are relevant and applicable to current research. It has generated a good deal of discussion about mitochondria and cell biology, and started conversations about students' choices for their own future careers. We are grateful to Jordan for taking the time to join us, and it was especially welcome at this time where our students' expected journey through their education has been disrupted. The A level students will be carrying out work based on what they learnt from Jordan's talk, and how it fits in with the work they do in A level Biology. |
| Year(s) Of Engagement Activity | 2020 |