PUBLICATION

Proteolysis regulates cardiomyocyte maturation and tissue integration

Authors

Fukuda, R., Gunawan, F., Beisaw, A., Jimenez-Amilburu, V., Maischein, H.M., Kostin, S., Kawakami, K., Stainier, D.Y.

ID

ZDB-PUB-170218-5

Date

2017

Source

Nature communications 8: 14495 (Journal)

Registered Authors

Kawakami, Koichi, Maischein, Hans-Martin, Stainier, Didier

Keywords

Cell biology, Developmental biology

Datasets

GEO:GSE85770

MeSH Terms

Rats
Mutation/genetics
Myocytes, Cardiac/metabolism*
Myocytes, Cardiac/pathology
Zebrafish/genetics
Zebrafish/growth & development*
Zebrafish/metabolism*
Organogenesis*
Animals
Mice
Proteolysis*
Cell Dedifferentiation
Animals, Newborn
Base Sequence
Heart Defects, Congenital/metabolism
Heart Defects, Congenital/pathology

(all 16)

PubMed

28211472 Full text @ Nat. Commun.

Abstract

Tissue integrity is critical for organ formation and function. During heart development, cardiomyocytes differentiate and integrate to form a coherent tissue that contracts synchronously. However, the molecular mechanisms regulating cardiac tissue integrity are poorly understood. Here we show that proteolysis, via the E3 ubiquitin ligase ASB2, regulates cardiomyocyte maturation and tissue integrity. Cardiomyocytes in asb2b zebrafish mutants fail to terminally differentiate, resulting in reduced cardiac contractility and output. Mosaic analyses reveal a cell-autonomous requirement for Asb2b in cardiomyocytes for their integration as asb2b mutant cardiomyocytes are unable to meld into wild-type myocardial tissue. In vitro and in vivo data indicate that ASB2 negatively regulates TCF3, a bHLH transcription factor. TCF3 must be degraded for cardiomyocyte maturation, as TCF3 gain-of-function causes a number of phenotypes associated with cardiomyocyte dedifferentiation. Overall, our results show that proteolysis has an important role in cardiomyocyte maturation and the formation of a coherent myocardial tissue.

Genes / Markers

Figures

Show all Figures

Expression

Gene	Fish	Conditions	Stage	Anatomy	Assay	Figure
asb2b	WT	standard conditions	Prim-5	heart tube	ISH	Fig. 1
asb2b	WT	standard conditions	Prim-5	somite	ISH	Fig. 1
EGFP	asb2b^bns33/bns33; bns103Tg; sd11Tg	standard conditions	Long-pec	cardiac muscle cell basal side	IFL	Fig. 2
EGFP	asb2b^bns33/bns33; s883Tg	standard conditions	Long-pec	cardiac muscle cell plasma membrane	IFL	Fig. 2
EGFP	asb2b^bns33/bns33; sd10Tg; sd11Tg	standard conditions	Long-pec	cardiac muscle cell Z disc	IFL	Fig. 2
EGFP	bns103Tg; sd11Tg	standard conditions	Long-pec	cardiac muscle cell apical side	IFL	Fig. 2
EGFP	bns106Tg; sd11Tg	standard conditions	Long-pec	cardiac muscle cell nucleus	IFL	Fig. 4
EGFP	bns107Tg; m1230Tg; sd11Tg	standard conditions	Long-pec	cardiac muscle cell	IFL	Fig. 1
EGFP	bns142Tg; s883Tg	standard conditions	Long-pec	cardiac muscle cell plasma membrane	IFL	Fig. 4
EGFP	bns142Tg; s974Tg	standard conditions	Long-pec	cardiac muscle cell myofilament	IFL	Fig. 4

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
asb2b^bns33/bns33	standard conditions	Long-pec	heart id2a expression increased amount, abnormal	Fig. 4
asb2b^bns33/bns33	standard conditions	Long-pec	heart mef2cb expression increased amount, abnormal	Fig. 4
asb2b^bns33/bns33	standard conditions	Long-pec	heart socs3b expression increased amount, abnormal	Fig. 4
asb2b^bns33/bns33	standard conditions	Long-pec	pericardium edematous, abnormal	Fig. 1
asb2b^bns33/bns33; bns103Tg; sd11Tg	standard conditions	Long-pec	cardiac muscle cell basal side EGFP expression mislocalised, abnormal	Fig. 2
asb2b^bns33/bns33; bns142Tg; s883Tg	standard conditions	Day 5	cardiac muscle cell plasma membrane has extra parts of type cardiac muscle cell cell projection, ameliorated	Fig. 4
asb2b^bns33/bns33; bns142Tg; s883Tg	standard conditions	Day 5	cardiac muscle cell shape, ameliorated	Fig. 4
asb2b^bns33/bns33; bns142Tg; s974Tg	standard conditions	Day 5	cardiac muscle cell myofilament disorganized, ameliorated	Fig. 4
asb2b^bns33/bns33; s883Tg	standard conditions	Long-pec	cardiac muscle cell plasma membrane has extra parts of type cardiac muscle cell cell projection, abnormal	Fig. 2
asb2b^bns33/bns33; s883Tg	standard conditions	Long-pec	cardiac muscle cell shape, abnormal	Fig. 2

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Mutations / Transgenics

Allele	Construct	Type	Affected Genomic Region
bns33		Small Deletion	asb2b
bns78Tg	Tg(myl7:cdh2-Tomato)	Transgenic Insertion
bns103Tg	Tg(myl7:EGFP-podxl)	Transgenic Insertion
bns106Tg	TgFOS(asb2b:GFP-asb2b)	Transgenic Insertion
bns107Tg	TgFOS(asb2b:GFP-GAL4FF)	Transgenic Insertion
bns141Tg	Tg(myl7:LIFEACT-Tomato)	Transgenic Insertion
bns142Tg	Tg(myl7:EGFP-tcf3b-2A-Tomato)	Transgenic Insertion
bns143Tg	Tg(myl7:id2b-2A-Tomato)	Transgenic Insertion
bns144		Insertion	asb2a.1
f2Tg	Tg(-5.1myl7:DsRed2-NLS)	Transgenic Insertion

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Human Disease / Model

Sequence Targeting Reagents

Target	Reagent	Reagent Type
asb2a.1	CRISPR1-asb2a.1	CRISPR
asb2b	CRISPR1-asb2b	CRISPR

Fish

Fish
asb2b^bns33/bns33
asb2b^bns33/bns33; bns103Tg; sd11Tg
asb2b^bns33/bns33; bns142Tg; s883Tg
asb2b^bns33/bns33; bns142Tg; s974Tg
asb2b^bns33/bns33; s883Tg
asb2b^bns33/bns33; s974Tg
asb2b^bns33/bns33; s974Tg; sd11Tg
asb2b^bns33/bns33; sd10Tg; sd11Tg
bns103Tg; sd11Tg
bns106Tg; sd11Tg

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Antibodies

Name	Type	Antigen Genes	Isotypes	Host Organism
Ab3-cdh2	polyclonal		IgG	Rabbit
Ab-MF20	monoclonal		IgG2b	Mouse

Orthology

Engineered Foreign Genes

Marker	Marker Type	Name
DsRed2	EFG	DsRed2
EGFP	EFG	EGFP
GAL4FF	EFG	GAL4FF
GCaMP	EFG	GCaMP
GFP	EFG	GFP
mKate2	EFG	mKate2
Tomato	EFG	Tomato

Mapping

Fukuda et al., 2017 ZDB-PUB-170218-5 PMID:28211472

Proteolysis regulates cardiomyocyte maturation and tissue integration

Fukuda et al., 2017

ZDB-PUB-170218-5
PMID:28211472