ERG activity is regulated by endothelial FAK coupling with TRIM25/USP9x in vascular patterning

D'Amico, G. et al. (2022) ERG activity is regulated by endothelial FAK coupling with TRIM25/USP9x in vascular patterning. Development, 149(13), dev200528. (doi: 10.1242/dev.200528) (PMID:35723257) (PMCID:PMC9340553)

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.

• Item Type: Articles
• Additional Information: GD’A was funded by Barts Charity (472/2174). IF, NB, JG-E were funded by Cancer Research UK (A18673). HK and PM were funded by (Breast Cancer Now as part of Programme Funding to the Breast Cancer Now Toby Robins Research Centre (CTR-QR14-007). We acknowledge NHS funding to the NIHR Biomedical Research Centre. EM, and JW are supported by (CRUK-Barts Cancer centre funds). LR was supported by MRC funds MR/V009621/1 and Cancer Research UK (A18673). MSA and FKM are funded and supported by a Medical Research Council Career Development Award (MR/P009417/1) and a Barts Charity grant (MGU0346) to FKM, and a Barry Read PhD studentship to MSA. RG and YW were supported by CRUK grant: A27224. KH-D is HEFCE funded and has had a Cancer Research UK programme grant (C82181/A12007) and recently (DRCNPG-May21\100004) which now supports LR. The laboratory was also supported by funds from Barts Charity (MGU0487) and the CRUK City of London Major Centre. AB-C Wellcome Trust (Multiuser Equipment 208402/Z/17). A Serrels, Cancer Research UK (grant number C39669/A25919). A Squire employed by the University Clinic Essen. B Serrels was HEFCE supported. GMB, British Heart Foundation Project Grant PG/17/33/32990. AMR, British Heart Foundation Programme Grant RG/17/4/32662.
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: Serrels, Dr Bryan
• Authors: D'Amico, G., Fernandez, I., Gómez-Escudero, J., Kim, H., Maniati, E., Azman, M. S., Mardakheh, F. K., Serrels, B., Serrels, A., Parsons, M., Squire, A., Birdsey, G. M., Randi, A. M., Bolado-Carrancio, A., Gangeswaran, R., Reynolds, L. E., Bodrug, N., Wang, Y., Wang, J., Meier, P., and Hodivala-Dilke, K. M.
• College/School: College of Medical Veterinary and Life Sciences > School of Cancer Sciences
• Journal Name: Development
• Publisher: The Company of Biologists
• ISSN: 1477-9129
• ISSN (Online): 1477-9129
• Published Online: 20 June 2022
• Copyright Holders: Copyright © 2022 The Company of Biologists
• First Published: First published in Development 149(13): dev200528
• Publisher Policy: Reproduced under a Creative Commons License