PUBLICATION

Spliceosomal protein eftud2 mutation leads to p53-dependent apoptosis in zebrafish neural progenitors

Authors

Lei, L., Yan, S.Y., Yang, R., Chen, J.Y., Li, Y., Bu, Y., Chang, N., Zhou, Q., Zhu, X., Li, C.Y., Xiong, J.W.

ID

ZDB-PUB-161203-16

Date

2017

Source

Nucleic acids research 45(6): 3422-3436 (Journal)

Registered Authors

Xiong, Jing-Wei

Keywords

none

Datasets

GEO:GSE78106

MeSH Terms

Animals
Apoptosis*
Brain/abnormalities
Cloning, Molecular
Exons
Introns
Mutation
Neural Stem Cells/cytology*
Neurogenesis/genetics*
Neurons/cytology
Nonsense Mediated mRNA Decay
Peptide Elongation Factors/genetics*
RNA Splicing
RNA Splicing Factors/genetics*
Spinal Cord/abnormalities
Transcriptome
Tumor Suppressor Protein p53/metabolism
Zebrafish/genetics
Zebrafish/growth & development
Zebrafish Proteins/genetics*
Zebrafish Proteins/metabolism

(all 21)

PubMed

27899647 Full text @ Nucleic Acids Res.

Abstract

Haploinsufficiency of EFTUD2 (Elongation Factor Tu GTP Binding Domain Containing 2) is linked to human mandibulofacial dysostosis, Guion-Almeida type (MFDGA), but the underlying cellular and molecular mechanisms remain to be addressed. We report here the isolation, cloning and functional analysis of the mutated eftud2 (snu114) in a novel neuronal mutant fn10a in zebrafish. This mutant displayed abnormal brain development with evident neuronal apoptosis while the development of other organs appeared less affected. Positional cloning revealed a nonsense mutation such that the mutant eftud2 mRNA encoded a truncated Eftud2 protein and was subjected to nonsense-mediated decay. Disruption of eftud2 led to increased apoptosis and mitosis of neural progenitors while it had little effect on differentiated neurons. Further RNA-seq and functional analyses revealed a transcriptome-wide RNA splicing deficiency and a large amount of intron-retaining and exon-skipping transcripts, which resulted in inadequate nonsense-mediated RNA decay and activation of the p53 pathway in fn10a mutants. Therefore, our study has established that eftud2 functions in RNA splicing during neural development and provides a suitable zebrafish model for studying the molecular pathology of the neurological disease MFDGA.

Genes / Markers

Marker	Marker Type	Name
aldoca	GENE	aldolase C, fructose-bisphosphate, a
atxn1b	GENE	ataxin 1b
baxa	GENE	BCL2 associated X, apoptosis regulator a
bbc3	GENE	BCL2 binding component 3
casp8	GENE	caspase 8, apoptosis-related cysteine peptidase
ccnb1	GENE	cyclin B1
cdkn1a	GENE	cyclin dependent kinase inhibitor 1A
diabloa	GENE	diablo, IAP-binding mitochondrial protein a
eftud2	GENE	elongation factor Tu GTP binding domain containing 2
foxa3	GENE	forkhead box A3

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Figures

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Expression

Gene	Antibody	Fish	Conditions	Stage	Qualifier	Anatomy	Assay	Figure
atxn1b		eftud2^fn10a/fn10a (TL)	standard conditions	Prim-25		whole organism	RTPCR	Fig. 6
atxn1b		TL + MO1-upf1	standard conditions	Prim-25		whole organism	RTPCR	Fig. 6
baxa		eftud2^fn10a/fn10a (TL)	standard conditions	Prim-25		whole organism	RTPCR	Fig. 7
bbc3		eftud2^fn10a/fn10a (TL)	standard conditions	Prim-25		whole organism	RTPCR	Fig. 7
casp8		eftud2^fn10a/fn10a (TL)	standard conditions	Prim-25		whole organism	RTPCR	Fig. 7
diabloa		eftud2^fn10a/fn10a (TL)	standard conditions	Prim-25		whole organism	RTPCR	Fig. 7
eftud2	Ab1-eftud2	eftud2^fn10a/fn10a (TL)	standard conditions	Prim-25	Not Detected	whole organism	WB	Fig. 2
eftud2		TL	standard conditions	1-cell	Not Detected	anatomical structure	ISH	Fig. 2
eftud2		TL	standard conditions	Prim-25		brain	ISH	Fig. 2
eftud2		TL	standard conditions	Prim-5		brain	ISH	Fig. 2

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
TL + MO1-eftud2	standard conditions	Prim-25	whole organism tp53 expression increased amount, abnormal	Fig. 7
TL + MO1-upf1	standard conditions	Prim-25	mRNA splicing, via spliceosome decreased occurrence, abnormal	Fig. 6
TL + MO1-upf1	standard conditions	Prim-25	nuclear-transcribed mRNA catabolic process, nonsense-mediated decay increased process quality, abnormal	Fig. 6
TL + MO1-upf1	standard conditions	Prim-25	whole organism atxn1b expression increased amount, abnormal	Fig. 6
TL + MO1-upf1	standard conditions	Prim-25	whole organism mmp30 expression increased amount, abnormal	Fig. 6
eftud2^fn10a/fn10a (TL)	standard conditions	Prim-25	brain apoptotic process increased occurrence, abnormal	Fig. 7
eftud2^fn10a/fn10a (TL)	standard conditions	Prim-25	brain apoptotic process increased occurrence, abnormal	Fig. 3
eftud2^fn10a/fn10a (TL)	standard conditions	Prim-25	brain cell population proliferation increased occurrence, abnormal	Fig. 3
eftud2^fn10a/fn10a (TL)	standard conditions	Prim-25	brain morphology, abnormal	Fig. 1
eftud2^fn10a/fn10a (TL)	standard conditions	Prim-25	brain neuron map2 expression decreased amount, abnormal	Fig. 1

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Mutations / Transgenics

Allele	Construct	Type	Affected Genomic Region
fn10a		Point Mutation	eftud2

Human Disease / Model

Human Disease	Fish	Conditions	Evidence
mandibulofacial dysostosis, Guion-Almeida type	eftud2^fn10a/fn10a (TL)	standard conditions	TAS
Treacher Collins syndrome	eftud2^fn10a/fn10a (TL)	standard conditions	TAS

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Sequence Targeting Reagents

Target	Reagent	Reagent Type
eftud2	MO1-eftud2	MRPHLNO
tp53	MO4-tp53	MRPHLNO
upf1	MO1-upf1	MRPHLNO

Fish

Fish
eftud2^fn10a/fn10a + MO4-tp53 (TL)
eftud2^fn10a/fn10a (TL)
TL
TL + MO1-eftud2
TL + MO1-upf1

Antibodies

Orthology

Gene	Orthology
eftud2

Engineered Foreign Genes

Mapping

Lei et al., 2017 ZDB-PUB-161203-16 PMID:27899647

Spliceosomal protein eftud2 mutation leads to p53-dependent apoptosis in zebrafish neural progenitors

Lei et al., 2017

ZDB-PUB-161203-16
PMID:27899647