PUBLICATION

Loss of zebrafish Smyd1a interferes with myofibrillar integrity without triggering the misfolded myosin response

Authors

Paone, C., Rudeck, S., Etard, C., Strähle, U., Rottbauer, W., Just, S.

ID

ZDB-PUB-180115-1

Date

2018

Source

Biochemical and Biophysical Research Communications 496(2): 339-345 (Journal)

Registered Authors

Etard, Christelle, Just, Steffen, Rottbauer, Wolfgang, Rudeck, Steven, Strähle, Uwe

Keywords

CRISPR/Cas9, Cardiomyocyte, Danio rerio, Misfolded myosin response, Skeletal muscle, Smyd1, mBop

MeSH Terms

Gene Expression Regulation, Developmental*
Animals, Genetically Modified
Morpholinos/genetics
Morpholinos/metabolism
Embryo, Nonmammalian
Muscle, Skeletal/metabolism*
Muscle, Skeletal/pathology
Histone-Lysine N-Methyltransferase/antagonists & inhibitors
Histone-Lysine N-Methyltransferase/deficiency
Histone-Lysine N-Methyltransferase/genetics*
HSP90 Heat-Shock Proteins/genetics
HSP90 Heat-Shock Proteins/metabolism
Green Fluorescent Proteins/genetics
Green Fluorescent Proteins/metabolism
Animals
Sarcomeres/metabolism*
Sarcomeres/pathology
Molecular Chaperones/genetics
Molecular Chaperones/metabolism
Gene Editing
Protein Folding
Myocytes, Cardiac/metabolism*
Myocytes, Cardiac/pathology
Humans
Zebrafish/genetics*
Zebrafish/growth & development
Zebrafish/metabolism
Genes, Reporter
Gene Duplication
CRISPR-Cas Systems
Myosins/genetics*
Myosins/metabolism
Protein Isoforms/deficiency
Protein Isoforms/genetics
Zebrafish Proteins/antagonists & inhibitors
Zebrafish Proteins/deficiency
Zebrafish Proteins/genetics*
Zebrafish Proteins/metabolism

(all 38)

PubMed

29331378 Full text @ Biochem. Biophys. Res. Commun.

Abstract

Sarcomeric protein turnover needs to be tightly balanced to assure proper assembly and renewal of sarcomeric units within muscle tissues. The mechanisms regulating these fundamental processes are only poorly understood, but of great clinical importance since many cardiac and skeletal muscle diseases are associated with defective sarcomeric organization. The SET- and MYND domain containing protein 1b (Smyd1b) is known to play a crucial role in myofibrillogenesis by functionally interacting with the myosin chaperones Unc45b and Hsp90α1. In zebrafish, Smyd1b, Unc45b and Hsp90α1 are part of the misfolded myosin response (MMR), a regulatory transcriptional response that is activated by disturbed myosin homeostasis. Genome duplication in zebrafish led to a second smyd1 gene, termed smyd1a. Morpholino- and CRISPR/Cas9-mediated knockdown of smyd1a led to significant perturbations in sarcomere structure resulting in decreased cardiac as well as skeletal muscle function. Similar to Smyd1b, we found Smyd1a to localize to the sarcomeric M-band in skeletal and cardiac muscles. Overexpression of smyd1a efficiently compensated for the loss of Smyd1b in flatline (fla) mutant zebrafish embryos, rescued the myopathic phenotype and suppressed the MMR in Smyd1b-deficient embryos, suggesting overlapping functions of both Smyd1 paralogs. Interestingly, Smyd1a is not transcriptionally activated in Smyd1b-deficient fla mutants, demonstrating lack of genetic compensation despite the functional redundancy of both zebrafish Smyd1 paralogs.

Genes / Markers

Figures

Show all Figures

Expression

Gene	Fish	Conditions	Stage	Anatomy	Assay	Figure
EGFP	ulm100Tg	standard conditions	Long-pec	skeletal muscle M band	IFL	Fig. 1
EGFP	ulm102Tg	standard conditions	Long-pec	skeletal muscle M band	IFL	Fig. 1
hsp90aa1.1	smyd1b^zf340/zf340	standard conditions	Protruding-mouth	skeletal muscle	ISH	Fig. 4
hsp90aa1.1	WT + MO4-smyd1a	standard conditions	Protruding-mouth	skeletal muscle	ISH	Fig. 4
smyd1a	smyd1b^zf340/zf340	standard conditions	Protruding-mouth	skeletal muscle	ISH	Fig. 3
smyd1a	unc45b^sb60/sb60	standard conditions	Protruding-mouth	skeletal muscle	ISH	Fig. 4
smyd1a	WT	standard conditions	Long-pec	fast muscle cell	ISH	Fig. 1
smyd1a	WT	standard conditions	Long-pec	heart	ISH	Fig. 1
smyd1a	WT	standard conditions	Long-pec	heart	RTPCR	Fig. 1
smyd1a	WT	standard conditions	Protruding-mouth	skeletal muscle	ISH	Fig. 3

1 - 10 of 16

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
WT + CRISPR2-smyd1a	standard conditions	Long-pec	skeletal muscle refractivity, abnormal	Fig. 2
WT + CRISPR2-smyd1a	standard conditions	Long-pec	thigmotaxis disrupted, abnormal	Fig. S1
WT + CRISPR2-smyd1a	standard conditions	Long-pec	thigmotaxis disrupted, abnormal	Fig. 2
WT + MO4-smyd1a	standard conditions	Prim-5	swimming process quality, normal	Fig. S1
WT + MO4-smyd1a	standard conditions	Long-pec	heart decreased functionality, abnormal	Fig. S1
WT + MO4-smyd1a	standard conditions	Long-pec	pericardium edematous, abnormal	Fig. 2
WT + MO4-smyd1a	standard conditions	Long-pec	skeletal muscle refractivity, abnormal	Fig. 2
WT + MO4-smyd1a	standard conditions	Long-pec	thigmotaxis disrupted, abnormal	Fig. 2
WT + MO5-smyd1a	standard conditions	Prim-5	swimming process quality, normal	Fig. S1
WT + MO5-smyd1a	standard conditions	Long-pec	heart decreased functionality, abnormal	Fig. S1

1 - 10 of 21

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Mutations / Transgenics

Allele	Construct	Type	Affected Genomic Region
sb60		Point Mutation	unc45b
ulm100Tg	Tg(unc45b:smyd1b-EGFP)	Transgenic Insertion
ulm102Tg	Tg(unc45b:smyd1a-EGFP)	Transgenic Insertion
zf340		Point Mutation	smyd1b

1 - 4 of 4

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Human Disease / Model

Sequence Targeting Reagents

Target	Reagent	Reagent Type
hsp90aa1.1	CRISPR1-hsp90aa1.1	CRISPR
smyd1a	CRISPR2-smyd1a	CRISPR
smyd1a	MO4-smyd1a	MRPHLNO
smyd1a	MO5-smyd1a	MRPHLNO

Fish

Fish
smyd1b^zf340/zf340
ulm100Tg
ulm102Tg
unc45b^sb60/sb60
WT
WT + CRISPR1-hsp90aa1.1
WT + CRISPR2-smyd1a
WT + MO4-smyd1a
WT + MO5-smyd1a

Antibodies

Orthology

Engineered Foreign Genes

Marker	Marker Type	Name
EGFP	EFG	EGFP

Mapping

Paone et al., 2018 ZDB-PUB-180115-1 PMID:29331378

Loss of zebrafish Smyd1a interferes with myofibrillar integrity without triggering the misfolded myosin response

Paone et al., 2018

ZDB-PUB-180115-1
PMID:29331378