PUBLICATION

The cerebral cavernous malformation proteins CCM2L and CCM2 prevent the activation of the MAP kinase MEKK3

Authors

Cullere, X., Plovie, E., Bennett, P.M., MacRae, C.A., Mayadas, T.N.

ID

ZDB-PUB-151106-1

Date

2015

Source

Proceedings of the National Academy of Sciences of the United States of America 112(46): 14284-9 (Journal)

Registered Authors

Keywords

MAP kinase, cerebral cavernous malformation, endothelium, expression, signaling

MeSH Terms

Multiprotein Complexes/genetics
Multiprotein Complexes/metabolism
Animals
Zebrafish/embryology*
Zebrafish/genetics
Mitogen-Activated Protein Kinase 7/genetics
Mitogen-Activated Protein Kinase 7/metabolism
Endothelial Cells/cytology
Endothelial Cells/metabolism
Cardiomegaly/embryology
Cardiomegaly/genetics
Transcription, Genetic/physiology*
MAP Kinase Signaling System/physiology*
MAP Kinase Kinase Kinase 3/genetics
MAP Kinase Kinase Kinase 3/metabolism*
Enzyme Activation/physiology
Gene Knockdown Techniques
Carrier Proteins/genetics
Carrier Proteins/metabolism*
Zebrafish Proteins/genetics
Zebrafish Proteins/metabolism*

(all 21)

PubMed

26540726 Full text @ Proc. Natl. Acad. Sci. USA

Abstract

Three genes, CCM1, CCM2, and CCM3, interact genetically and biochemically and are mutated in cerebral cavernous malformations (CCM). A recently described member of this CCM family of proteins, CCM2-like (CCM2L), has high homology to CCM2. Here we show that its relative expression in different tissues differs from that of CCM2 and, unlike CCM2, the expression of CCM2L in endothelial cells is regulated by density, flow, and statins. In vitro, both CCM2L and CCM2 bind MEKK3 in a complex with CCM1. Both CCM2L and CCM2 interfere with MEKK3 activation and its ability to phosphorylate MEK5, a downstream target. The in vivo relevance of this regulation was investigated in zebrafish. A knockdown of ccm2l and ccm2 in zebrafish leads to a more severe "big heart" and circulation defects compared with loss of function of ccm2 alone, and also leads to substantial body axis abnormalities. Silencing of mekk3 rescues the big heart and body axis phenotype, suggesting cross-talk between the CCM proteins and MEKK3 in vivo. In endothelial cells, CCM2 deletion leads to activation of ERK5 and a transcriptional program that are downstream of MEKK3. These findings suggest that CCM2L and CCM2 cooperate to regulate the activity of MEKK3.

Genes / Markers

Figures

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Expression

Phenotype

Fish	Conditions	Stage	Phenotype	Figure
AB/TU + MO1-map3k3 + MO3-ccm2l	standard conditions	Long-pec	heart increased size, ameliorated	Fig. 3
AB/TU + MO1-map3k3 + MO3-ccm2l	standard conditions	Long-pec	whole organism anterior-posterior axis deformed, ameliorated	Fig. 3
AB/TU + MO4-ccm2l	standard conditions	Long-pec	heart increased size, abnormal	Fig. S2
ccm2^m201/+ + MO1-map3k3 + MO3-ccm2l	standard conditions	Long-pec	heart increased size, ameliorated	Fig. 3
ccm2^m201/+ + MO1-map3k3 + MO3-ccm2l	standard conditions	Long-pec	whole organism anterior-posterior axis deformed, ameliorated	Fig. 3
ccm2^m201/+ + MO1-tnnt2a	standard conditions	Long-pec	heart increased size, abnormal	Fig. S2
ccm2^m201/+ + MO1-tnnt2a	standard conditions	Long-pec	whole organism anterior-posterior axis morphology, normal	Fig. S2
ccm2^m201/+ + MO3-ccm2l	standard conditions	Long-pec	heart increased size, abnormal	Fig. 3
ccm2^m201/+ + MO3-ccm2l	standard conditions	Long-pec	heart non-functional, abnormal	Fig. 3
ccm2^m201/+ + MO3-ccm2l	standard conditions	Long-pec	whole organism anterior-posterior axis deformed, abnormal	Fig. 3

1 - 10 of 23

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Mutations / Transgenics

Allele	Construct	Type	Affected Genomic Region
la116Tg	Tg(kdrl:GFP)	Transgenic Insertion
m201		Point Mutation	ccm2
m775		Point Mutation	krit1

Human Disease / Model

Sequence Targeting Reagents

Target	Reagent	Reagent Type
ccm2l	MO3-ccm2l	MRPHLNO
ccm2l	MO4-ccm2l	MRPHLNO
map3k3	MO1-map3k3	MRPHLNO
tnnt2a	MO1-tnnt2a	MRPHLNO

Fish

Fish
AB/TU + MO1-map3k3 + MO3-ccm2l
AB/TU + MO4-ccm2l
ccm2^m201/+ + MO1-map3k3 + MO3-ccm2l
ccm2^m201/+ + MO1-tnnt2a
ccm2^m201/+ + MO3-ccm2l
ccm2^m201/+ + MO4-ccm2l
krit1^m775/+ + MO3-ccm2l
la116Tg + MO3-ccm2l

1 - 8 of 8

Show

Antibodies

Orthology

Gene	Orthology
ccm2l

Engineered Foreign Genes

Marker	Marker Type	Name
GFP	EFG	GFP

Mapping

Cullere et al., 2015 ZDB-PUB-151106-1 PMID:26540726

The cerebral cavernous malformation proteins CCM2L and CCM2 prevent the activation of the MAP kinase MEKK3

Cullere et al., 2015

ZDB-PUB-151106-1
PMID:26540726