PUBLICATION

Zebrafish rbm8a and magoh mutants reveal EJC developmental functions and new 3'UTR intron-containing NMD targets

Authors

Gangras, P., Gallagher, T.L., Parthun, M.A., Yi, Z., Patton, R.D., Tietz, K.T., Deans, N.C., Bundschuh, R., Amacher, S.L., Singh, G.

ID

ZDB-PUB-200606-31

Date

2020

Source

PLoS Genetics 16: e1008830 (Journal)

Registered Authors

Amacher, Sharon, Gallagher, Thomas, Tietz, Kiel

Keywords

none

MeSH Terms

Zebrafish/genetics
Zebrafish/growth & development
Axons/physiology
Humans
RNA-Binding Proteins/genetics
RNA-Binding Proteins/metabolism*
Neuronal Outgrowth/genetics
Exons/genetics
Mutation
Gene Expression Regulation, Developmental*
Introns/genetics
RNA, Messenger/metabolism
RNA-Seq
Codon, Terminator
Embryo, Nonmammalian
Sequence Alignment
Datasets as Topic
Homozygote
3' Untranslated Regions/genetics
Nuclear Proteins/genetics
Forkhead Transcription Factors/metabolism*
Nonsense Mediated mRNA Decay/genetics*
Animals
Mice
Zebrafish Proteins/metabolism*
Peptide Chain Termination, Translational
Muscle, Skeletal/innervation
Mutagenesis
Animals, Genetically Modified
Neurogenesis/genetics*
Gene Regulatory Networks/genetics
Up-Regulation

(all 32)

PubMed

32502192 Full text @ PLoS Genet.

Abstract

Many post-transcriptional mechanisms operate via mRNA 3'UTRs to regulate protein expression, and such controls are crucial for development. We show that homozygous mutations in two zebrafish exon junction complex (EJC) core genes rbm8a and magoh leads to muscle disorganization, neural cell death, and motor neuron outgrowth defects, as well as dysregulation of mRNAs subjected to nonsense-mediated mRNA decay (NMD) due to translation termination ≥ 50 nts upstream of the last exon-exon junction. Intriguingly, we find that EJC-dependent NMD also regulates a subset of transcripts that contain 3'UTR introns (3'UI) < 50 nts downstream of a stop codon. Some transcripts containing such stop codon-proximal 3'UI are also NMD-sensitive in cultured human cells and mouse embryonic stem cells. We identify 167 genes that contain a conserved proximal 3'UI in zebrafish, mouse and humans. foxo3b is one such proximal 3'UI-containing gene that is upregulated in zebrafish EJC mutant embryos, at both mRNA and protein levels, and loss of foxo3b function in EJC mutant embryos significantly rescues motor axon growth defects. These data are consistent with EJC-dependent NMD regulating foxo3b mRNA to control protein expression during zebrafish development. Our work shows that the EJC is critical for normal zebrafish development and suggests that proximal 3'UIs may serve gene regulatory function in vertebrates.

Genes / Markers

Figures

Show all Figures

Expression

Phenotype

Fish	Conditions	Stage	Phenotype	Figure
WT	chemical treatment by environment: inhibitor	Prim-5	whole organism atxn1b expression increased amount, abnormal	Fig 6
WT	chemical treatment by environment: inhibitor	Prim-5	whole organism cdkn1ba expression increased amount, abnormal	Fig 6
WT	chemical treatment by environment: inhibitor	Prim-5	whole organism eif4a2 expression increased amount, abnormal	Fig 6
WT	chemical treatment by environment: inhibitor	Prim-5	whole organism foxo3b expression increased amount, abnormal	Fig 6
WT	chemical treatment by environment: inhibitor	Prim-5	whole organism mob4 expression amount, ameliorated	Fig 6
WT	chemical treatment by environment: inhibitor	Prim-5	whole organism phlda2 expression increased amount, abnormal	Fig 6
magoh^oz37/oz37	standard conditions	14-19 somites	muscle contraction decreased occurrence, abnormal	Fig 3
magoh^oz37/oz37	standard conditions	14-19 somites	whole organism paralysed, abnormal	Fig 3
magoh^oz37/oz37	standard conditions	20-25 somites	whole organism atxn1b expression increased amount, abnormal	Fig 5
magoh^oz37/oz37	standard conditions	20-25 somites	whole organism eif4a2 expression increased amount, abnormal	Fig 5

1 - 10 of 52

Show

Mutations / Transgenics

Allele	Type	Affected Genomic Region
ihb404	Insertion	foxo3b
oz36	Small Deletion	rbm8a
oz37	Small Deletion	magoh

Human Disease / Model

Sequence Targeting Reagents

Target	Reagent	Reagent Type
magoh	CRISPR1-magoh	CRISPR
rbm8a	CRISPR1-rbm8a	CRISPR
upf1	MO2-upf1	MRPHLNO

Fish

Fish
magoh^oz37/oz37
magoh^oz37/oz37; foxo3b^{ihb404/ihb404}
magoh^oz37/oz37; foxo3b^ihb404/+
rbm8a^oz36/oz36
WT

Antibodies

Orthology

Engineered Foreign Genes

Mapping

Gangras et al., 2020 ZDB-PUB-200606-31 PMID:32502192

Zebrafish rbm8a and magoh mutants reveal EJC developmental functions and new 3'UTR intron-containing NMD targets

Gangras et al., 2020

ZDB-PUB-200606-31
PMID:32502192