Project

Back to overview

Elucidating the biological function of FUS and its role in neurodegeneration

Applicant Mühlemann Oliver
Number 136222
Funding scheme Sinergia
Research institution Departement für Chemie und Biochemie Universität Bern
Institution of higher education University of Berne - BE
Main discipline Molecular Biology
Start/End 01.09.2011 - 30.11.2014
Approved amount 1'515'400.00
Show all

All Disciplines (5)

Discipline
Molecular Biology
Biophysics
Cellular Biology, Cytology
Neurophysiology and Brain Research
Biochemistry

Keywords (12)

RNA-protein interaction; RNA metabolism; FUS; neurodegeneration; miRNA biogenesis; pre-mRNA splicing; genome stability; amyotrophic lateral sclerosis; transgenic mouse model; NMR; ALS; TLS

Lay Summary (English)

Lead
Lay summary

Ribonucleic acid (RNA) plays a central role in the expression of genetic information both as an information carrier (messenger RNA [mRNA]) and as a regulator (micro-RNA [miRNA]). Reflecting its important role for life, uncontrolled alterations of RNA metabolism often leads to disease. For example, research during the last years revealed that mutations in proteins with roles in RNA metabolism are implicated in a wide variety of neurodegenerative diseases, but the pathogenic mechanisms leading to these disorders are not yet understood. This research project focuses on the RNA-binding protein FUS (fused in sarcoma, also known as translated in liposarcoma, TLS) and aims at elucidating its biological function(s).  Mutations in the FUS gene have recently been identified in patients with an inherited form of amyotrophic lateral sclerosis (ALS). Most of these mutations lead to cytoplasmic accumulation of FUS aggregates in neurons and glial cells of these ALS patients. Moreover, such FUS aggregates are also the characteristic hallmark lesions in a subset of patients with frontotemporal lobar degeneration (FTLD).

FUS is a ubiquitously expressed protein with several structural motifs in the C-terminal half that appear to be involved in nucleic acid binding. Consistent with this, FUS has been implicated in a variety of cellular processes linked to RNA metabolism, miRNA maturation, and DNA repair, but its exact function is not yet clear. With an interdisciplinary team of experts in both RNA biology and molecular pathology of neurodegenerative disorders, and with a broad spectrum of methods, comprising structural biology, biochemistry, molecular and cell biology, and transgenic mouse models, we plan to i) identify genome-wide the RNAs and proteins interacting with FUS, ii) determine the structural basis for the specificity of these interactions, iii) understand the molecular role of FUS in selective miRNA maturation, regulation of alternative splicing, and DNA damage response, and iv) find out how a failure in any of these potential functions of FUS can lead to neurodegeneration and specifically to death of motor neurons.

The results of this project are expected to not only reveal the molecular mechanism leading to FUS-associated ALS and FTLD, but generally to provide significant new insight into the emerging connection between misregulation of RNA metabolism and neurodegeneration, a connection that is currently implied in a variety of additional neurological diseases. Understanding these molecular mechanisms will serve as the basis for future development of effective therapies.

Direct link to Lay Summary Last update: 21.02.2013

Responsible applicant and co-applicants

Employees

Publications

Publication
The Solution Structure of FUS Bound to RNA Reveals a Bipartite Mode of RNA Recognition with Both Sequence and Shape Specificity
Loughlin Fionna E., Lukavsky Peter J., Kazeeva Tamara, Reber Stefan, Hock Eva-Maria, Colombo Martino, Von Schroetter Christine, Pauli Phillip, Cléry Antoine, Mühlemann Oliver, Polymenidou Magdalini, Ruepp Marc-David, Allain Frédéric H.-T. (2018), The Solution Structure of FUS Bound to RNA Reveals a Bipartite Mode of RNA Recognition with Both Sequence and Shape Specificity, in Molecular Cell, 73, 1-15.
Minor intron splicing is regulated by FUS and affected by ALS-associated FUS mutants
Reber Stefan, Stettler Jolanda, Filosa Giuseppe, Colombo Martino, Jutzi Daniel, Lenzken Silvia, Schweingruber Christoph, Bruggmann Remy, Bachi Angela, Barabino Silvia, Mühlemann Oliver, Ruepp Marc-David (2016), Minor intron splicing is regulated by FUS and affected by ALS-associated FUS mutants, in EMBO Journal, 35(14), 1504-1521.
Effect of combined systemic and local morpholino treatment on the spinal muscular atrophy δ7 mouse model phenotype
Nizzardo Monica, Simone Chiara, Salani Sabrina, Ruepp Marc David, Rizzo Federica, Ruggieri Margherita, Zanetta Chiara, Brajkovic Simona, Moulton Hong M., Muehlemann Oliver, Bresólin Nereo, Comi Giacomo Pietro, Corti Stefania (2014), Effect of combined systemic and local morpholino treatment on the spinal muscular atrophy δ7 mouse model phenotype, in Clinical Therapeutics, 36(3), 340-356.
Neuronal RNA-binding proteins in health and disease
Lenzken Silvia Carolina, Achsel Tilmann, Carri Maria Teresa, Barabino Silvia ML (2014), Neuronal RNA-binding proteins in health and disease, in WIREs RNA, 5(4), 565.
Frontotemporal lobar degeneration and amyotrophic lateral sclerosis: Molecular similarities and differences.
Neumann M (2013), Frontotemporal lobar degeneration and amyotrophic lateral sclerosis: Molecular similarities and differences., in Revue neurologique, 169(10), 793-798.
Molecular basis of UG-rich RNA recognition by the human splicing factor TDP-43
Lukavsky Peter J., Daujotyte Dalia, Tollervey James R., Ule Jernej, Stuani Cristiana, Buratti Emanuele, Baralle Francisco E., Damberger Fred F., Allain Frederic H-T (2013), Molecular basis of UG-rich RNA recognition by the human splicing factor TDP-43, in NATURE STRUCTURAL & MOLECULAR BIOLOGY, 20(12), 1443-1449.
Paraquat Modulates Alternative Pre-mRNA Splicing by Modifying the Intracellular Distribution of SRPK2
Vivarelli Silvia, Lenzken Silvia, Ruepp Marc-David, Ranzini Francesco, Maffioletti Andrea, Alvarez Reinaldo, Mühlemann Oliver, Barabino Silvia (2013), Paraquat Modulates Alternative Pre-mRNA Splicing by Modifying the Intracellular Distribution of SRPK2, in PLoS one, 8(4), e61980.
Truncating mutations in FUS/TLS give rise to a more aggressive ALS-phenotype than missense mutations: A clinico-genetic study in Germany
Waibel Stefan, Neumann Manuela, Rosenbohm Angela, Birve Anna, Volk Alexander E., Weishaupt Jochen H., Meyer Thomas P H, Müller Ulrich F., Andersen Peter M., Ludolph Albert Christian (2013), Truncating mutations in FUS/TLS give rise to a more aggressive ALS-phenotype than missense mutations: A clinico-genetic study in Germany, in European Journal of Neurology, 20(3), 540-546.
Advances in understanding the molecular basis of frontotemporal dementia
Rademakers R., Neumann M., Mackenzie I. R. (2012), Advances in understanding the molecular basis of frontotemporal dementia, in Nat Rev Neurol, 8, 423-34.
Mechanisms of disease in frontotemporal lobar degeneration: gain of function versus loss of function effects
Halliday G., Bigio E. H., Cairns N. J., Neumann M., Mackenzie I. R., Mann D. M. (2012), Mechanisms of disease in frontotemporal lobar degeneration: gain of function versus loss of function effects, in Acta Neuropathol, 124, 373-82.
Characterizing new fluorescent tools for studying 5-HT3 receptor pharmacology
Jack Thomas, Simonin Jonathan, Ruepp Marc-David, Thompson Andrew J, Gertsch Jürg, Lochner Martin, Characterizing new fluorescent tools for studying 5-HT3 receptor pharmacology, in Neuropharmacology, 90, 63.

Collaboration

Group / person Country
Types of collaboration
Neuralstem Lab Stefania Corti, Univ. of Milano Italy (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Exchange of personnel
Istituto di Ricerche Farmacologiche "Mario Negri" Italy (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Research Infrastructure
Eva Hedlund, Karolinska Institutet, Stockholm Sweden (Europe)
- in-depth/constructive exchanges on approaches, methods or results
Mass spectrometry lab Angela Bachi, San Raffaele, Milano Italy (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Research Infrastructure
Daniel Schümperli, IZB, Uni Bern Switzerland (Europe)
- in-depth/constructive exchanges on approaches, methods or results
R. Colagero, Univ. Turin Italy (Europe)
- in-depth/constructive exchanges on approaches, methods or results
Genecore facility, EMBL Heidelberg Germany (Europe)
- Research Infrastructure
Martin Lochner, DCB, Uni Bern Switzerland (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Research Infrastructure
- Exchange of personnel

Scientific events

Active participation

Title Type of contribution Title of article or contribution Date Place Persons involved
Symposium “RNA meets DNA: on the road to genome instability” Poster Investigating the role of the RNA-binding protein FUS in the DNA damage response 03.11.2014 Baeza, Spain Lenzken Silvia Carolina;
9th International Conference on Frontotemporal Dementias Talk given at a conference Clinico-Pathological Aspects of FTLD-TDP and FTLD-FET 23.10.2014 Vancouver, Canada Neumann Manuela;
TERM 2014 Talk given at a conference The FUS(s) about splicing 22.10.2014 Paris, France Ruepp Marc-David;
Annual Swiss RNA Workshop 2013 Poster The multifunctional FUS protein: Characterization of the biological effects of its depletion 25.01.2014 bern, Switzerland Barabino Silvia; Lenzken Silvia Carolina; Mühlemann Oliver; Ruepp Marc-David;
Annual Swiss RNA Workshop 2014 Poster Characterization of the role of the RNA-binding protein FUS in the DNA damage response 24.01.2014 Bern, Switzerland Ruepp Marc-David; Barabino Silvia; Lenzken Silvia Carolina;
Annual Swiss RNA Workshop 2014 Poster Proteomic characterization of the FUS/TLS interactome 24.01.2014 Bern, Switzerland Mühlemann Oliver; Barabino Silvia; Ruepp Marc-David; Lenzken Silvia Carolina;
Annual Swiss RNA Workshop 2014 Talk given at a conference The role of FUS in splicing regulation 24.01.2014 Bern, Switzerland Ruepp Marc-David; Mühlemann Oliver;
Annual Swiss RNA Workshop 2014 Poster Genetic Engineering of Induced Pluripotent Stem Cells and Reprogramming to Motor Neurons to Elucidate Selective Motor Neuron Death in Amyotrophic Lateral Sclerosis 24.01.2014 Bern, Switzerland Mühlemann Oliver; Ruepp Marc-David;
18th Annual meeting of the RNA society Poster The FUS protein is required for cell proliferation 11.06.2013 Davos, Switzerland Barabino Silvia; Lenzken Silvia Carolina; Ruepp Marc-David; Mühlemann Oliver;
18th Annual meeting of the RNA society Poster Characterization of the potential role for RNA-binding protein FUS in DNA damage response: A quantitative proteomic approach 11.06.2013 Davos, Switzerland Lenzken Silvia Carolina; Ruepp Marc-David; Mühlemann Oliver; Barabino Silvia;
18th Annual meeting of the RNA society Talk given at a conference The SWI/SNF subunit Brahma modulates the choice of alternative terminal exons by recruiting the BRCA1/BARD1 ubiquitin ligase 11.06.2013 Davos, Switzerland Barabino Silvia; Lenzken Silvia Carolina;
18th Annual meeting of the RNA society Poster The involvement of miRNA Dysregulation in Amyotrophic Lateral Sclerosis 11.06.2013 Davos, Switzerland Mühlemann Oliver; Ruepp Marc-David; Barabino Silvia;
18th Annual meeting of the RNA society Poster FUS protein interacts with U7 snRNP and plays a role in replication-dependant histone genes expression 11.06.2013 Davos, Switzerland Ruepp Marc-David;
18th Annual meeting of the RNA society Poster Combined systemic and local morpholino treatment rescues the phenotype of the SMA Delta 7 mouse model 11.06.2013 Davos, Switzerland Mühlemann Oliver; Ruepp Marc-David;
11th International Conference on Alzheimer's and Parkinson's disease Talk given at a conference TDP-43, FUS and Beyond: Insights on the Role of RNA-binding Proteins in Frontotemporal Dementia and Amyotrophic Lateral Sclerosis 06.03.2013 Florence, Italy Neumann Manuela;
Annual Swiss RNA Workshop 2013 Poster Unravelling the multiple roles of FUS in RNA processing and on ALS pathogenesis 25.01.2013 Bern, Switzerland Ruepp Marc-David; Mühlemann Oliver;
Annual Swiss RNA Workshop 2013 Poster Unravelling the impact of microRNA on Amyotrophic Lateral Sclerosis (ALS) pathogenesis 25.01.2013 Bern, Switzerland Barabino Silvia; Ruepp Marc-David;
Annual Swiss RNA Workshop 2013 Poster Characterization of the potential role for RNA-binding protein FUS/TLS in DNA damage response: A quantitative proteomic approach 25.01.2013 Bern, Switzerland Mühlemann Oliver; Ruepp Marc-David; Barabino Silvia; Lenzken Silvia Carolina;
Neurex WORKSHOP ON “NEW BREAKTHROUGHS IN MOTOR NEURON DISEASE" Talk given at a conference FUS, a link between amyotrophic lateral sclerosis and frontotemporal lobar degeneration 12.11.2012 Strasbourg, France Neumann Manuela;
8th international Conference on Frontotemporal Dementias Talk given at a conference Mechanisms of Disease in FTLD – a gain, or a loss of function effect? 05.09.2012 Manchester, UK, Great Britain and Northern Ireland Neumann Manuela;
SIBBM "Frontiers in Molecular Biology" Seminar 2013, Revisiting the Central Dogma: Emerging New Concepts in Replication, Transcription, and Translation Talk given at a conference Oxidative stress alters the choice of alternative last exons via Brahma downregulation and disassembly of the pre-mRNA 3' end processing factor CstF 04.06.2012 Pavia, Italy Fontana Gabriele A.;
Gordon Conference on Chromatin Structure & Function Poster The chromatin-remodelling factor Brahma modulates the choice of alternative terminal exons 06.05.2012 Lucca, Italy Fontana Gabriele A.;
1st Post-EURASNET Symposium on Regulation of Gene Expression through RNA Splicing Talk given at a conference The chromatin-remodelling factor Brahma modulates the choice of alternative terminal exons 24.03.2012 Trieste, Italy Barabino Silvia;
Annual Swiss RNA Workshop 2012 Talk given at a conference The chromatin-remodelling factor Brahma modulates the choice of alternative terminal exons 03.02.2012 Bern, Switzerland Barabino Silvia;
Annual Swiss RNA Workshop 2012 Poster Unravelling the multiple roles of FUS in RNA processing and the impact of microRNAs on ALS pathogenesis 03.02.2012 Bern, Switzerland Ruepp Marc-David; Mühlemann Oliver;
Symposium “Frontiers in Epigenomics” Talk given at a conference The chromatin-remodelling factor Brahma modulates the choice of alternative terminal 17.10.2011 Baeza, Spain Barabino Silvia;
Symposium “The enemy within: endogenous DNA damage as a source of cancer and ageing” Poster Genotoxic stress modulates alternative pre-mRNA splicing by modifying the intracellular distribution of SRPK2 17.10.2011 Baeza, Spain Lenzken Silvia Carolina;


Awards

Title Year
Prize of Italian Society for Biochemistry and Molecular Biology for best oral presentation at the annual meeting 2013

Associated projects

Number Title Start Funding scheme
157704 Direct electron detector and phase plate for cryo-transmission electron microscopy of biological samples 01.12.2014 R'EQUIP
149921 NMR structure determination of protein-RNA complexes involved in pre-mRNA splicing and translation regulation 01.10.2013 Project funding (Div. I-III)

Abstract

Mutations in proteins with roles in RNA metabolism are implicated in a wide variety of neurodegenerative diseases, but the pathogenic mechanisms leading to these disorders are not well understood. For example, mutations in the FUS gene (fused in sarcoma; also known as translated in liposarcoma, TLS), which encodes an RNA-binding protein of the hnRNP family, have recently been identified in patients with an inherited form of amyotrophic lateral sclerosis (ALS). Most of these mutations destroy the nuclear localization signal (NLS) of the normally predominantly nuclear FUS protein, leading to cytoplasmic accumulation of FUS aggregates in neurons and glial cells of these ALS patients. Moreover, FUS positive inclusions are also the characteristic hallmark lesions in a subset of patients with frontotemporal lobar degeneration. It is currently unclear if the FUS mutations-associated form of ALS develops due to a gain-of-function of the mutated protein (i.e. toxicity of the cytoplasmatic FUS aggregates), due to a loss-of-function (i.e. inhibition of FUS’ nuclear function), or due to a combination of both. In this Sinergia project, we propose to test the loss-of-function hypothesis. To this end, we want to elucidate the biological function of FUS, which so far is not well understood.FUS is a ubiquitously expressed protein with several structural motifs in the C-terminal half that appear to be involved in nucleic acid binding. Consistent with this, FUS has been implicated in a variety of cellular processes linked to RNA metabolism, miRNA maturation, and DNA repair. With an interdisciplinary team of experts in both RNA biology (F. Allain, S. Barabino, O. Mühlemann) and molecular pathology of neurodegenerative disorders (M. Neumann), and with a broad spectrum of methods, comprising structural biology, biochemistry, molecular and cell biology, and transgenic mouse models, we plan to i) identify genome-wide the RNAs and proteins interacting with FUS, ii) determine the structural basis for the specificity of these interactions, iii) understand the molecular role of FUS in selective miRNA maturation, regulation of alternative splicing, and DNA damage response, and iv) find out how a failure in any of these potential functions of FUS can lead to neurodegeneration and specifically to death of motor neurons.The results of our research are expected to not only reveal the molecular mechanism leading to FUS-associated ALS and FTLD, but generally to provide significant new insight into the emerging connection between misregulation of RNA metabolism and neurodegeneration, a connection that is currently implied in a variety of addiditional neurological diseases, including spinocerebellar ataxia 2 (SCA-2), spinal muscular atrophy (SMA), fragile X syndrome, and myotonic dystrophy. Understanding these molecular mechanisms will serve as the basis for future development of effective therapies.
-