ERG activity is regulated by endothelial FAK coupling with TRIM25/USP9x in vascular patterning
File(s)dev200528.pdf (6.7 MB)
Accepted version
Author(s)
Type
Journal Article
Abstract
Precise vascular patterning is critical for normal growth and development. The ERG transcription factor drives Delta like ligand 4 (DLL4)/Notch signalling and is thought to act as pivotal regulators of endothelial cell (EC) dynamics and developmental angiogenesis. However, molecular regulation of ERG activity remains obscure. Using a series of EC specific Focal Adhesion Kinase (FAK)-knockout (KO) and point-mutant FAK-knockin mice, we show that loss of ECFAK, its kinase activity or phosphorylation at FAK-Y397, but not FAK-Y861, reduces ERG and DLL4 expression levels together with concomitant aberrations in vascular patterning. Rapid Immunoprecipitation Mass Spectrometry of Endogenous Proteins identified that endothelial nuclear-FAK interacts with the de-ubiquitinase USP9x and the ubiquitin ligase TRIM25 enzymes. Further in silico analysis corroborates that ERG interacts with USP9x and TRIM25. Moreover, ERG levels are reduced in FAKKO ECs via a ubiquitin-mediated post-translational modification programme involving USP9x and TRIM25. Re-expression of ERG in vivo and in vitro rescues the aberrant vessel sprouting defects observed in the absence of ECFAK. Our findings identify ECFAK as a regulator of retinal vascular patterning by controlling ERG protein degradation via TRIM25/USP9x.
Date Issued
2022-06-20
Date Acceptance
2022-04-29
Citation
Development, 2022, 149 (13)
ISSN
0950-1991
Publisher
Company of Biologists Ltd
Journal / Book Title
Development
Volume
149
Issue
13
Copyright Statement
© 2022 The Author(s). Published by The Company of Biologists Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.
License URL
Sponsor
British Heart Foundation
British Heart Foundation
Identifier
https://www.ncbi.nlm.nih.gov/pubmed/35723257
PII: 275750
Grant Number
PG/17/33/32990
RG/11/17/29256
Subjects
Endothelial cell signaling
Retinal angiogenesis
Vascular patterning regulation
Publication Status
Published online
Coverage Spatial
England
Date Publish Online
2022-06-20