PUBLICATION

Spliceosomal components protect embryonic neurons from R-loop-mediated DNA damage and apoptosis

Authors

Sorrells, S., Nik, S., Casey, M., Cameron, R.C., Truong, H., Toruno, C., Gulfo, M., Lowe, A., Jette, C., Stewart, R.A., Bowman, T.V.

ID

ZDB-PUB-180209-5

Date

2018

Source

Disease models & mechanisms 11(2): (Journal)

Registered Authors

Bowman, Teresa, Jette, Cicely A., Sorrells, Shelly, Stewart, Rodney A., Toruno, Cristhian, Truong, Harold

Keywords

Apoptosis, Neurons, R-loops, Radiation, Splicing, Zebrafish

MeSH Terms

Spliceosomes/metabolism*
Neurons/cytology*
Neurons/metabolism*
Neurons/radiation effects
Tumor Suppressor Protein p53/metabolism
Cytoprotection*/radiation effects
RNA Splicing/genetics
RNA Splicing/radiation effects
Zebrafish Proteins/metabolism
Zebrafish/embryology
Zebrafish/genetics*
Radiation Tolerance/genetics
Radiation Tolerance/radiation effects
DNA Breaks, Double-Stranded
Radiation, Ionizing
Genes, Essential
DNA Damage*
Nucleic Acid Conformation*
Mutation/genetics
Apoptosis*/radiation effects
Animals

(all 21)

PubMed

29419415 Full text @ Dis. Model. Mech.

Abstract

RNA splicing factors are essential for the viability of all eukaryotic cells; however, in metazoans some cell types are exquisitely sensitive to disruption of splicing factors. Neuronal cells represent one such cell type, and defects in RNA splicing factors can lead to neurodegenerative diseases. The basis for this tissue selectivity is not well understood owing to difficulties in analyzing the consequences of splicing factor defects in whole-animal systems. Here, we use zebrafish mutants to show that loss of spliceosomal components, including splicing factor 3b, subunit 1 (sf3b1), causes increased DNA double-strand breaks and apoptosis in embryonic neurons. Moreover, these mutants show a concomitant accumulation of R-loops, which are non-canonical nucleic acid structures that promote genomic instability. Dampening R-loop formation by conditional induction of ribonuclease H1 in sf3b1 mutants reduced neuronal DNA damage and apoptosis. These findings show that splicing factor dysfunction leads to R-loop accumulation and DNA damage that sensitizes embryonic neurons to apoptosis. Our results suggest that diseases associated with splicing factor mutations could be susceptible to treatments that modulate R-loop levels.

Genes / Markers

Figures

Show all Figures

Expression

Gene	Fish	Conditions	Stage	Anatomy	Assay	Figure
tp53	hspa8^hi138Tg (AB)	standard conditions	Prim-15	whole organism	WB	Fig. 2
tp53	plrg1^hi3174aTg (AB)	standard conditions	Prim-15	whole organism	WB	Fig. 2
tp53	sf3b1^{hi3394aTg/hi3394aTg} (AB)	standard conditions	Prim-15	whole organism	WB	Fig. 2
tp53	yju2^{zd1000/zd1000} (AB)	standard conditions	Prim-15	whole organism	WB	Fig. 2

1 - 4 of 4

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
AB + MO1-plrg1	electromagnetic radiation	Prim-5	apoptotic process increased occurrence, abnormal	Fig. 1
AB + MO1-plrg1	electromagnetic radiation	Prim-5	whole organism increased sensitivity to irradiation, abnormal	Fig. 1
hspa8^hi138Tg (AB)	standard conditions	Prim-15	whole organism tp53 expression increased amount, abnormal	Fig. 2
plrg1^hi3174aTg (AB)	standard conditions	Prim-15	whole organism tp53 expression increased amount, abnormal	Fig. 2
plrg1^hi3174aTg (AB)	standard conditions	Prim-15	whole organism ab2-h2afx labeling increased amount, abnormal	Fig. 2
sf3b1^{hi3394aTg/hi3394aTg} (AB)	standard conditions	Prim-5	apoptotic process increased occurrence, abnormal	Fig. 4
sf3b1^{hi3394aTg/hi3394aTg} (AB)	standard conditions	Prim-5	whole organism ab7-h2afx labeling increased amount, abnormal	Fig. 4
sf3b1^{hi3394aTg/hi3394aTg} (AB)	standard conditions	Prim-15	whole organism tp53 expression increased amount, abnormal	Fig. 2
sf3b1^{hi3394aTg/hi3394aTg} (AB)	standard conditions	Prim-15	whole organism ab2-h2afx labeling increased amount, abnormal	Fig. 2
sf3b1^{hi3394aTg/hi3394aTg} (AB)	electromagnetic radiation	Prim-5	apoptotic process increased occurrence, abnormal	Fig. 1

1 - 10 of 31

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Mutations / Transgenics

Human Disease / Model

Sequence Targeting Reagents

Target	Reagent	Reagent Type
plrg1	MO1-plrg1	MRPHLNO

Fish

Fish
AB + MO1-plrg1
hspa8^hi138Tg (AB)
plrg1^hi3174aTg (AB)
sf3b1^{hi3394aTg/hi3394aTg} (AB)
sf3b1^{hi3394aTg/hi3394aTg}; zd1009Tg (AB)
sfpq^{hi1779Tg/hi1779Tg} (AB)
tp53^zdf1/zdf1; plrg1^hi3174aTg (AB)
tp53^zdf1/zdf1; sf3b1^{hi3394aTg/hi3394aTg} (AB)
tp53^zdf1/zdf1; yju2^{zd1000/zd1000} (AB)
txnl4a^{zd1006/zd1006} (AB)

1 - 10 of 11

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Antibodies

Orthology

Engineered Foreign Genes

Marker	Marker Type	Name
GFP	EFG	GFP

Mapping

Sorrells et al., 2018 ZDB-PUB-180209-5 PMID:29419415

Spliceosomal components protect embryonic neurons from R-loop-mediated DNA damage and apoptosis

Sorrells et al., 2018

ZDB-PUB-180209-5
PMID:29419415