Differential role for rapid proteostasis in Rho GTPase-mediated control of quiescent endothelial integrity

Fabienne Podieh, Roos Wensveen, Max C. Overboom, Lotte Abbas, Jisca Majolée, Peter L. Hordijk*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review


Endothelial monolayer permeability is regulated by actin dynamics and vesicular traffic. Recently, ubiquitination was also implicated in the integrity of quiescent endothelium, as it differentially controls the localization and stability of adhesion and signaling proteins. However, the more general effect of fast protein turnover on endothelial integrity is not clear. Here, we found that inhibition of E1 ubiquitin ligases induces a rapid, reversible loss of integrity in quiescent, primary human endothelial monolayers, accompanied by increased F-actin stress fibers and the formation of intercellular gaps. Concomitantly, total protein and activity of the actin-regulating GTPase RhoB, but not its close homolog RhoA, increase ∼10-fold in 5 to 8 h. We determined that the depletion of RhoB, but not of RhoA, the inhibition of actin contractility, and the inhibition of protein synthesis all significantly rescue the loss of cell–cell contact induced by E1 ligase inhibition. Collectively, our data suggest that in quiescent human endothelial cells, the continuous and fast turnover of short-lived proteins that negatively regulate cell–cell contact is essential to preserve monolayer integrity.
Original languageEnglish
Article number104593
JournalJournal of Biological Chemistry
Issue number4
Publication statusPublished - 1 Apr 2023

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