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

• DOI: 10.1371/journal.pgen.1008830
• Publication Date: 2020-06-05T17:54:07Z
• Authors: Pooja Gangras; Thomas L. Gallagher; Michael A. Parthun; Zhongxia Yi; Robert D. Patton; Kiel T. Tietz; Natalie C. Deans; Ralf Bundschuh; Sharon L. Amacher; Guramrit Singh
• Keywords: 0301 basic medicine; Embryo, Nonmammalian; Neurogenesis; Neuronal Outgrowth; Datasets as Topic; QH426-470; Animals, Genetically Modified; Mice; 03 medical and health sciences; Genetics; Animals; Humans; Gene Regulatory Networks; Muscle, Skeletal; 3' Untranslated Regions; 0303 health sciences; Homozygote; Gene Expression Regulation, Developmental; Forkhead Transcription Factors; Exons; Axons; Introns; 3. Good health; Mutagenesis; Mutation; Codon, Terminator; Research Article

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.