Myoferlin regulates vascular endothelial growth factor receptor-2 stability and function.
Myoferlin and dysferlin are members of the ferlin family of membrane proteins. Recent studies have shown that mutation or genetic disruption of myoferlin or dysferlin promotes muscular dystrophy-related phenotypes in mice, which are the result of impaired plasma membrane integrity. However, no biological functions have been ascribed to myoferlin in non-muscle tissues. Herein, using a proteomic analysis of endothelial cell (EC) caveolae/lipid raft microdomains we identified myoferlin in these domains and show that myoferlin is highly expressed in ECs and vascular tissues. The loss of myoferlin results in lack of proliferation, migration, and nitric oxide (NO) release in response to vascular endothelial growth factor (VEGF). Western blotting and surface biotinylation experiments show that loss of myoferlin reduces the expression level and autophosphorylation of VEGF receptor-2 (VEGFR-2) in native ECs. In a reconstituted cell system, transfection of myoferlin increases VEGFR-2 membrane expression and autophosphorylation in response to VEGF. In vivo, VEGFR-2 levels and VEGF-induced permeability are impaired in myoferlin-deficient mice. Mechanistically, myoferlin forms a complex with dynamin-2 and VEGFR-2, which prevents CBL-dependent VEGFR-2 polyubiquitination and proteasomal degradation. These data are the first to report novel biological activities for myoferlin and reveal the role of membrane integrity to VEGF signaling.
Bernatchez
Pascal N
PN
Department of Pharmacology and Vascular Biology & Transplantation Program, Boyer Center for Molecular Medicine, Yale University School of Medicine, New Haven, Connecticut 06536, USA.
Acevedo
Lisette
L
Fernandez-Hernando
Carlos
C
Murata
Takahisa
T
Chalouni
Cecile
C
Kim
Jiae
J
Erdjument-Bromage
Hediye
H
Shah
Vijay
V
Gratton
Jean-Philippe
JP
McNally
Elizabeth M
EM
Tempst
Paul
P
Sessa
William C
WC
D000818:Animals; D002135:Calcium-Binding Proteins; D002199:Capillary Permeability / Q000187:drug effects / Q000235:genetics; D021941:Caveolae / Q000378:metabolism* / Q000473:pathology; D002460:Cell Line; D002465:Cell Movement / Q000187:drug effects / Q000235:genetics; D049109:Cell Proliferation / Q000187:drug effects; D034285:Dynamin II / Q000235:genetics / Q000378:metabolism; D000073939:Dysferlin; D042783:Endothelial Cells / Q000378:metabolism* / Q000473:pathology; D017353:Gene Deletion; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D018345:Mice, Knockout; D009124:Muscle Proteins / Q000235:genetics / Q000378:metabolism*; D009137:Muscular Dystrophy, Animal / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D009569:Nitric Oxide / Q000235:genetics / Q000378:metabolism; D009928:Organ Specificity / Q000187:drug effects / Q000235:genetics; D010641:Phenotype; D010766:Phosphorylation / Q000187:drug effects; D046988:Proteasome Endopeptidase Complex / Q000235:genetics / Q000378:metabolism; D011499:Protein Processing, Post-Translational / Q000187:drug effects / Q000235:genetics; D040901:Proteomics; D050721:Proto-Oncogene Proteins c-cbl / Q000235:genetics / Q000378:metabolism; D025801:Ubiquitin / Q000235:genetics / Q000378:metabolism; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism / Q000494:pharmacology; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000235:genetics / Q000378:metabolism*
D002135:Calcium-Binding Proteins; C114839:DYSF protein, human; C497505:Dysf protein, mouse; D000073939:Dysferlin; C415584:MYOF protein, human; D008565:Membrane Proteins; D009124:Muscle Proteins; D025801:Ubiquitin; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; C521810:myoferlin protein, mouse; C467480:vascular endothelial growth factor A, mouse; D009569:Nitric Oxide; D050721:Proto-Oncogene Proteins c-cbl; D040301:Vascular Endothelial Growth Factor Receptor-2; D046988:Proteasome Endopeptidase Complex; D034285:Dynamin II; C067419:CBL protein, human; C493849:Cbl protein, mouse