Nuclear import factor transportin and arginine methyltransferase 1 modify FUS neurotoxicity in Drosophila

Autoren

Sandra Jäckel
Anna K Summerer
Catherine M Thömmes
Xia Pan
Aaron Voigt
Jörg B Schulz
Tobias M Rasse
Dorothee Dormann
Christian Haass
Philipp J Kahle

DOI

10.1016/j.nbd.2014.11.003

ISSN

0969-9961

Zeitschrift

Neurobiology of Disease

Sprache

en

Paginierung

76 - 88

Datum der Veröffentlichung

2015

Status

Published

Herausgeber

Elsevier BV

Herausgeber URL

http://dx.doi.org/10.1016/j.nbd.2014.11.003

Datum der Datenerfassung

2019

Titel

Nuclear import factor transportin and arginine methyltransferase 1 modify FUS neurotoxicity in Drosophila

Ausgabe der Zeitschrift

74

Data source: Crossref

Abstract

Inclusions containing Fused in Sarcoma (FUS) are found in familial and sporadic cases of the incurable progressive motor neuron disease amyotrophic lateral sclerosis and in a common form of dementia, frontotemporal dementia. Most disease-associated mutations are located in the C-terminal proline-tyrosine nuclear localization sequence (PY-NLS) of FUS and impair its nuclear import. It has been shown in cell culture that the nuclear import of FUS is mediated by transportin, which binds the PY-NLS and the last arginine/glycine/glycine-rich (RGG) domain of FUS. Methylation of this last RGG domain by protein arginine methyltransferases (PRMTs) weakens transportin binding and therefore impairs nuclear translocation of FUS. To investigate the requirements for the nuclear import of FUS in an in vivo model, we generated different transgenic Drosophila lines expressing human FUS wild type (hFUS wt) and two disease-related variants P525L and R495X, in which the NLS is mutated or completely absent, respectively. To rule out effects caused by heterologous hFUS expression, we analysed the corresponding variants for the Drosophila FUS orthologue Cabeza (Caz wt, P398L, Q349X). Expression of these variants in eyes and motor neurons confirmed the PY-NLS-dependent nuclear localization of FUS/Caz and caused neurodegenerative effects. Surprisingly, FUS/Caz toxicity was correlated to the degree of its nuclear localization in this overexpression model. High levels of nuclear FUS/Caz became insoluble and reduced the endogenous Caz levels, confirming FUS autoregulation in Drosophila. RNAi-mediated knockdown of the two transportin orthologues interfered with the nuclear import of FUS/Caz and also enhanced the eye phenotype. Finally, we screened the Drosophila PRMT proteins (DART1-9) and found that knockdown of Dart1 led to a reduction in methylation of hFUS P525L and aggravated its phenotype. These findings show that the molecular mechanisms controlling the nuclear import of FUS/Caz and FUS autoregulation are conserved between humans and Drosophila. In addition to the well-known neurodegenerative effects of FUS loss-of function, our data suggest toxic potential of overexpressed FUS in the nucleus and of insoluble FUS.

Addresses

German Center for Neurodegenerative Diseases (DZNE) Tübingen, Germany; Hertie Institute for Clinical Brain Research, Laboratory of Functional Neurogenetics, Tübingen, Germany. Electronic address: sandra.jaeckel@dzne.de.

Autoren

Sandra Jäckel
Anna K Summerer
Catherine M Thömmes
Xia Pan
Aaron Voigt
Jörg B Schulz
Tobias M Rasse
Dorothee Dormann
Christian Haass
Philipp J Kahle

DOI

10.1016/j.nbd.2014.11.003

eISSN

1095-953X

Externe Identifier

PubMed Identifier: 25447237

Funding acknowledgements

Deutsche Forschungsgemeinschaft: KA1675/3-1 and -2
German Competence Network “Degenerative Dementias”: 01GI1005B
DZNE:
Hertie Foundation:

Open access

false

ISSN

0969-9961

Zeitschrift

Neurobiology of disease

Schlüsselwörter

Eye
Motor Neurons
Animals
Animals, Genetically Modified
Humans
Drosophila melanogaster
Movement Disorders
Neurodegenerative Diseases
Disease Models, Animal
Methyltransferases
Karyopherins
RNA-Binding Proteins
RNA-Binding Protein FUS
Transcription Factor TFIID
Drosophila Proteins
DNA Methylation
RNA Interference
Active Transport, Cell Nucleus
Mutation
Gene Knockdown Techniques

Sprache

eng

Medium

Print-Electronic

Online publication date

2014

Paginierung

76 - 88

Datum der Veröffentlichung

2015

Status

Published

Datum der Datenerfassung

2014

Titel

Nuclear import factor transportin and arginine methyltransferase 1 modify FUS neurotoxicity in Drosophila.

Sub types

Research Support, Non-U.S. Gov't
Journal Article

Ausgabe der Zeitschrift

74

Data source: Europe PubMed Central

Abstract

Inclusions containing Fused in Sarcoma (FUS) are found in familial and sporadic cases of the incurable progressive motor neuron disease amyotrophic lateral sclerosis and in a common form of dementia, frontotemporal dementia. Most disease-associated mutations are located in the C-terminal proline-tyrosine nuclear localization sequence (PY-NLS) of FUS and impair its nuclear import. It has been shown in cell culture that the nuclear import of FUS is mediated by transportin, which binds the PY-NLS and the last arginine/glycine/glycine-rich (RGG) domain of FUS. Methylation of this last RGG domain by protein arginine methyltransferases (PRMTs) weakens transportin binding and therefore impairs nuclear translocation of FUS. To investigate the requirements for the nuclear import of FUS in an in vivo model, we generated different transgenic Drosophila lines expressing human FUS wild type (hFUS wt) and two disease-related variants P525L and R495X, in which the NLS is mutated or completely absent, respectively. To rule out effects caused by heterologous hFUS expression, we analysed the corresponding variants for the Drosophila FUS orthologue Cabeza (Caz wt, P398L, Q349X). Expression of these variants in eyes and motor neurons confirmed the PY-NLS-dependent nuclear localization of FUS/Caz and caused neurodegenerative effects. Surprisingly, FUS/Caz toxicity was correlated to the degree of its nuclear localization in this overexpression model. High levels of nuclear FUS/Caz became insoluble and reduced the endogenous Caz levels, confirming FUS autoregulation in Drosophila. RNAi-mediated knockdown of the two transportin orthologues interfered with the nuclear import of FUS/Caz and also enhanced the eye phenotype. Finally, we screened the Drosophila PRMT proteins (DART1-9) and found that knockdown of Dart1 led to a reduction in methylation of hFUS P525L and aggravated its phenotype. These findings show that the molecular mechanisms controlling the nuclear import of FUS/Caz and FUS autoregulation are conserved between humans and Drosophila. In addition to the well-known neurodegenerative effects of FUS loss-of function, our data suggest toxic potential of overexpressed FUS in the nucleus and of insoluble FUS.

Date of acceptance

2014

Autoren

Sandra Jäckel
Anna K Summerer
Catherine M Thömmes
Xia Pan
Aaron Voigt
Jörg B Schulz
Tobias M Rasse
Dorothee Dormann
Christian Haass
Philipp J Kahle

Autoren-URL

https://www.ncbi.nlm.nih.gov/pubmed/25447237

DOI

10.1016/j.nbd.2014.11.003

eISSN

1095-953X

Zeitschrift

Neurobiol Dis

Schlüsselwörter

ALS
Caz
Drosophila
FTLD
FUS
PRMT
Transportin
Active Transport, Cell Nucleus
Animals
Animals, Genetically Modified
DNA Methylation
Disease Models, Animal
Drosophila Proteins
Drosophila melanogaster
Eye
Gene Knockdown Techniques
Humans
Karyopherins
Methyltransferases
Motor Neurons
Movement Disorders
Mutation
Neurodegenerative Diseases
RNA Interference
RNA-Binding Protein FUS
RNA-Binding Proteins
Transcription Factor TFIID

Sprache

eng

Country

United States

Paginierung

76 - 88

PII

S0969-9961(14)00341-6

Datum der Veröffentlichung

2015

Status

Published

Datum, an dem der Datensatz öffentlich gemacht wurde

2016

Titel

Nuclear import factor transportin and arginine methyltransferase 1 modify FUS neurotoxicity in Drosophila.

Sub types

Journal Article
Research Support, Non-U.S. Gov't

Ausgabe der Zeitschrift

74

Data source: PubMed

Nuclear import factor transportin and arginine methyltransferase 1 modify FUS neurotoxicity in Drosophila

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