Full UPF3B function is critical for neuronal differentiation of neural stem cells

Tahani Alrahbeni, Francesca Sartor, Jihan Anderson, Zosia Miedzybrodzka, Colin McCaig, Berndt Muller

Research output: Contribution to journalArticle

18 Citations (Scopus)
5 Downloads (Pure)

Abstract

Background

Mutation in the UPF3B gene on chromosome X is implicated in neurodevelopmental disorders including X-linked intellectual disability, autism and schizophrenia. The protein UPF3B is involved in the nonsense-mediated mRNA decay pathway (NMD) that controls mRNA stability and functions in the prevention of the synthesis of truncated proteins.

Results

Here we show that NMD pathway components UPF3B and UPF1 are down-regulated during differentiation of neural stem cells into neurons. Using tethered function assays we found that UPF3B missense mutations described in families with neurodevelopmental disorders reduced the activity of UPF3B protein in NMD. In neural stem cells, UPF3B protein was detected in the cytoplasm and in the nucleus. Similarly in neurons, UPF3B protein was detected in neurites, the somatic cytoplasm and in the nucleus. In both cell types nuclear UPF3B protein was enriched in the nucleolus. Using GFP tagged UPF3B proteins we found that the missense mutations did not affect the cellular localisation. Expression of missense mutant UPF3B disturbed neuronal differentiation and reduced the complexity of the branching of neurites. Neuronal differentiation was similarly affected in the presence of the NMD inhibitor Amlexanox. The expression of mutant UPF3B proteins lead to a subtle increase in mRNA levels of selected NMD targets.

Conclusions

Together our findings indicate that, despite the down-regulation of NMD factors, functional NMD is critical for neuronal differentiation. We propose that the neurodevelopmental phenotype of UPF3B missense mutation is caused by impairment of NMD function altering neuronal differentiation.
Original languageEnglish
Article number33
Number of pages15
JournalMolecular brain
Volume8
Early online date27 May 2015
DOIs
Publication statusPublished - 27 May 2015

Fingerprint

Nonsense Mediated mRNA Decay
Neural Stem Cells
Missense Mutation
Proteins
Neurites
Cytoplasm
Neurons
X-Linked Genes
Critical Pathways
RNA Stability
Mutant Proteins
Autistic Disorder
Nuclear Proteins
Intellectual Disability
Schizophrenia
Down-Regulation
Phenotype
Messenger RNA

Keywords

  • autism
  • schizophrenia
  • X-linked intellectual disability
  • nonsense-mediated mRNA decay
  • UPF3B mutation
  • UPF1
  • tethered function assay
  • qPCR
  • Arhgap24
  • Atf4
  • protein localisation

Cite this

Full UPF3B function is critical for neuronal differentiation of neural stem cells. / Alrahbeni, Tahani; Sartor, Francesca; Anderson, Jihan; Miedzybrodzka, Zosia; McCaig, Colin; Muller, Berndt.

In: Molecular brain, Vol. 8, 33, 27.05.2015.

Research output: Contribution to journalArticle

@article{43e52a861cfc4377932a437556564af7,
title = "Full UPF3B function is critical for neuronal differentiation of neural stem cells",
abstract = "BackgroundMutation in the UPF3B gene on chromosome X is implicated in neurodevelopmental disorders including X-linked intellectual disability, autism and schizophrenia. The protein UPF3B is involved in the nonsense-mediated mRNA decay pathway (NMD) that controls mRNA stability and functions in the prevention of the synthesis of truncated proteins. ResultsHere we show that NMD pathway components UPF3B and UPF1 are down-regulated during differentiation of neural stem cells into neurons. Using tethered function assays we found that UPF3B missense mutations described in families with neurodevelopmental disorders reduced the activity of UPF3B protein in NMD. In neural stem cells, UPF3B protein was detected in the cytoplasm and in the nucleus. Similarly in neurons, UPF3B protein was detected in neurites, the somatic cytoplasm and in the nucleus. In both cell types nuclear UPF3B protein was enriched in the nucleolus. Using GFP tagged UPF3B proteins we found that the missense mutations did not affect the cellular localisation. Expression of missense mutant UPF3B disturbed neuronal differentiation and reduced the complexity of the branching of neurites. Neuronal differentiation was similarly affected in the presence of the NMD inhibitor Amlexanox. The expression of mutant UPF3B proteins lead to a subtle increase in mRNA levels of selected NMD targets. ConclusionsTogether our findings indicate that, despite the down-regulation of NMD factors, functional NMD is critical for neuronal differentiation. We propose that the neurodevelopmental phenotype of UPF3B missense mutation is caused by impairment of NMD function altering neuronal differentiation.",
keywords = "autism, schizophrenia, X-linked intellectual disability, nonsense-mediated mRNA decay, UPF3B mutation, UPF1, tethered function assay, qPCR, Arhgap24, Atf4, protein localisation",
author = "Tahani Alrahbeni and Francesca Sartor and Jihan Anderson and Zosia Miedzybrodzka and Colin McCaig and Berndt Muller",
note = "Acknowledgments We thank Fred H Gage (Salk Institute, La Jolla, CA, USA) for HCN-A94 cells and Niels Gehring (University of Cologne, Germany) for constructs. We gratefully acknowledge Tenovus Scotland (Project Grant G11-06), Moonlight Prowl (FS) and the Saudi Arabian Ministry of Higher Education via King Abdullah Program for Scholarships for support (TA). JA is supported by a PhD studentship from Medical Research Scotland (PhD-654-2012) and Dundee Cell Products.",
year = "2015",
month = "5",
day = "27",
doi = "10.1186/s13041-015-0122-1",
language = "English",
volume = "8",
journal = "Molecular brain",
issn = "1756-6606",
publisher = "BioMed Central",

}

TY - JOUR

T1 - Full UPF3B function is critical for neuronal differentiation of neural stem cells

AU - Alrahbeni, Tahani

AU - Sartor, Francesca

AU - Anderson, Jihan

AU - Miedzybrodzka, Zosia

AU - McCaig, Colin

AU - Muller, Berndt

N1 - Acknowledgments We thank Fred H Gage (Salk Institute, La Jolla, CA, USA) for HCN-A94 cells and Niels Gehring (University of Cologne, Germany) for constructs. We gratefully acknowledge Tenovus Scotland (Project Grant G11-06), Moonlight Prowl (FS) and the Saudi Arabian Ministry of Higher Education via King Abdullah Program for Scholarships for support (TA). JA is supported by a PhD studentship from Medical Research Scotland (PhD-654-2012) and Dundee Cell Products.

PY - 2015/5/27

Y1 - 2015/5/27

N2 - BackgroundMutation in the UPF3B gene on chromosome X is implicated in neurodevelopmental disorders including X-linked intellectual disability, autism and schizophrenia. The protein UPF3B is involved in the nonsense-mediated mRNA decay pathway (NMD) that controls mRNA stability and functions in the prevention of the synthesis of truncated proteins. ResultsHere we show that NMD pathway components UPF3B and UPF1 are down-regulated during differentiation of neural stem cells into neurons. Using tethered function assays we found that UPF3B missense mutations described in families with neurodevelopmental disorders reduced the activity of UPF3B protein in NMD. In neural stem cells, UPF3B protein was detected in the cytoplasm and in the nucleus. Similarly in neurons, UPF3B protein was detected in neurites, the somatic cytoplasm and in the nucleus. In both cell types nuclear UPF3B protein was enriched in the nucleolus. Using GFP tagged UPF3B proteins we found that the missense mutations did not affect the cellular localisation. Expression of missense mutant UPF3B disturbed neuronal differentiation and reduced the complexity of the branching of neurites. Neuronal differentiation was similarly affected in the presence of the NMD inhibitor Amlexanox. The expression of mutant UPF3B proteins lead to a subtle increase in mRNA levels of selected NMD targets. ConclusionsTogether our findings indicate that, despite the down-regulation of NMD factors, functional NMD is critical for neuronal differentiation. We propose that the neurodevelopmental phenotype of UPF3B missense mutation is caused by impairment of NMD function altering neuronal differentiation.

AB - BackgroundMutation in the UPF3B gene on chromosome X is implicated in neurodevelopmental disorders including X-linked intellectual disability, autism and schizophrenia. The protein UPF3B is involved in the nonsense-mediated mRNA decay pathway (NMD) that controls mRNA stability and functions in the prevention of the synthesis of truncated proteins. ResultsHere we show that NMD pathway components UPF3B and UPF1 are down-regulated during differentiation of neural stem cells into neurons. Using tethered function assays we found that UPF3B missense mutations described in families with neurodevelopmental disorders reduced the activity of UPF3B protein in NMD. In neural stem cells, UPF3B protein was detected in the cytoplasm and in the nucleus. Similarly in neurons, UPF3B protein was detected in neurites, the somatic cytoplasm and in the nucleus. In both cell types nuclear UPF3B protein was enriched in the nucleolus. Using GFP tagged UPF3B proteins we found that the missense mutations did not affect the cellular localisation. Expression of missense mutant UPF3B disturbed neuronal differentiation and reduced the complexity of the branching of neurites. Neuronal differentiation was similarly affected in the presence of the NMD inhibitor Amlexanox. The expression of mutant UPF3B proteins lead to a subtle increase in mRNA levels of selected NMD targets. ConclusionsTogether our findings indicate that, despite the down-regulation of NMD factors, functional NMD is critical for neuronal differentiation. We propose that the neurodevelopmental phenotype of UPF3B missense mutation is caused by impairment of NMD function altering neuronal differentiation.

KW - autism

KW - schizophrenia

KW - X-linked intellectual disability

KW - nonsense-mediated mRNA decay

KW - UPF3B mutation

KW - UPF1

KW - tethered function assay

KW - qPCR

KW - Arhgap24

KW - Atf4

KW - protein localisation

U2 - 10.1186/s13041-015-0122-1

DO - 10.1186/s13041-015-0122-1

M3 - Article

VL - 8

JO - Molecular brain

JF - Molecular brain

SN - 1756-6606

M1 - 33

ER -