RhoA effectors ROCK and SLK/LOK oppositely regulate VE-cadherin mobility at endothelial junctions
Abstract
Cellular contractile forces regulate VE-cadherin dynamics, adhesion, and endothelial junction integrity. RhoA controls these forces through two parallel effector kinases: ROCK, which activates non-muscle myosin II motors, and SLK/LOK, which promote actin-membrane attachment through phosphorylation of ezrin, radixin, moesin proteins (ERM). How these two RhoA effector pathways differentially affect VE-cadherin dynamics at junctions is not known. Using single-molecule tracking of Halo-tagged VE-cadherin, we obtained a detailed characterization of VE-cadherin diffusive behavior inside and outside junctional regions in cultured human umbilical vein endothelial cells (HUVEC). VE-cadherin was substantially less mobile and more confined within junctions than in non-junctional membrane. Inhibiting ROCK with Y-27632 increased VE-cadherin mobility both inside and outside junctions, consistent with RhoA/ROCK-driven contractility restraining cadherin diffusion globally. Unexpectedly, inhibiting SLK/LOK produced the opposite effect, selectively reducing VE-cadherin mobility within junctions with no comparable change outside them. This effect was accompanied by an increase in reticular junctions, suggesting that SLK/LOK inhibition alters VE-cadherin mobility indirectly, through a change in junctional architecture, rather than through a direct effect of actin-membrane coupling on VE-cadherin itself. These findings identify SLK/LOK as a regulator of VE-cadherin dynamics acting in parallel to ROCK downstream of RhoA and point to junctional architecture as a previously unappreciated determinant of VE-cadherin mobility at endothelial cell-cell contacts.
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BibTeX
@misc{nelson2026rhoa,
title = {{RhoA effectors ROCK and SLK/LOK oppositely regulate VE-cadherin mobility at endothelial junctions}},
author = {Tyler Nelson and Qiyao Lin and Zoya Quddoos and Henry Gatica-Gutierrez and Sofia Vargas-Hernandez and Arnold Hayer and Anna-Karin Gustavsson},
howpublished = {bioRxiv},
year = {2026},
month = oct,
doi = {10.64898/2026.10.28.762575},
url = {https://www.biorxiv.org/content/10.64898/2026.10.28.762575v1},
note = {CC BY-NC-ND},
}
RIS
TY - UNPB
TI - RhoA effectors ROCK and SLK/LOK oppositely regulate VE-cadherin mobility at endothelial junctions
AU - Tyler Nelson
AU - Qiyao Lin
AU - Zoya Quddoos
AU - Henry Gatica-Gutierrez
AU - Sofia Vargas-Hernandez
AU - Arnold Hayer
AU - Anna-Karin Gustavsson
PY - 2026
DA - 2026/10/28
PB - bioRxiv
DO - 10.64898/2026.10.28.762575
UR - https://www.biorxiv.org/content/10.64898/2026.10.28.762575v1
AB - Cellular contractile forces regulate VE-cadherin dynamics, adhesion, and endothelial junction integrity. RhoA controls these forces through two parallel effector kinases: ROCK, which activates non-muscle myosin II motors, and SLK/LOK, which promote actin-membrane attachment through phosphorylation of ezrin, radixin, moesin proteins (ERM). How these two RhoA effector pathways differentially affect VE-cadherin dynamics at junctions is not known. Using single-molecule tracking of Halo-tagged VE-cadherin, we obtained a detailed characterization of VE-cadherin diffusive behavior inside and outside junctional regions in cultured human umbilical vein endothelial cells (HUVEC). VE-cadherin was substantially less mobile and more confined within junctions than in non-junctional membrane. Inhibiting ROCK with Y-27632 increased VE-cadherin mobility both inside and outside junctions, consistent with RhoA/ROCK-driven contractility restraining cadherin diffusion globally. Unexpectedly, inhibiting SLK/LOK produced the opposite effect, selectively reducing VE-cadherin mobility within junctions with no comparable change outside them. This effect was accompanied by an increase in reticular junctions, suggesting that SLK/LOK inhibition alters VE-cadherin mobility indirectly, through a change in junctional architecture, rather than through a direct effect of actin-membrane coupling on VE-cadherin itself. These findings identify SLK/LOK as a regulator of VE-cadherin dynamics acting in parallel to ROCK downstream of RhoA and point to junctional architecture as a previously unappreciated determinant of VE-cadherin mobility at endothelial cell-cell contacts.
SN - 2692-8205
ER -