RhoGTPases control endothelial cell (EC) migration, adhesion, and barrier formation. Whereas the relevance of RhoA for endothelial barrier function is widely accepted, the role of the RhoA homologue RhoB is poorly defined. RhoB and RhoA are 85% identical, but RhoB's subcellular localization and half-life are uniquely different. Here, we studied the role of ubiquitination for the function and stability of RhoB in primary human ECs. We show that the K63 polyubiquitination at lysine 162 and 181 of RhoB targets the protein to lysosomes. Moreover, we identified the RING E3 ligase complex Cullin-3-Rbx1-KCTD10 as key modulator of endothelial barrier integrity via its regulation of the ubiquitination, localization, and activity of RhoB. In conclusion, our data show that ubiquitination controls the subcellular localization and lysosomal degradation of RhoB and thereby regulates the stability of the endothelial barrier through control of RhoBmediated EC contraction.

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Persistent URL dx.doi.org/10.1083/jcb.201606055, hdl.handle.net/1765/105058
Journal The Journal of Cell Biology
Rights no subscription
Citation
Kovačević, I, Sakaue, T. (Tomohisa), Majolee, J, Pronk, M.C. (Manon C.), Maekawa, M, Geerts, D, … Hordijk, P.L. (2018). The Cullin-3-Rbx1-KCTD10 complex controls endothelial barrier function via K63 ubiquitination of RhoB. The Journal of Cell Biology, 217(3), 1015–1032. doi:10.1083/jcb.201606055