A mechanism of Rap1-induced stabilization of endothelial cell--cell junctions.

Abstract:

:Activation of Rap1 small GTPases stabilizes cell--cell junctions, and this activity requires Krev Interaction Trapped gene 1 (KRIT1). Loss of KRIT1 disrupts cardiovascular development and causes autosomal dominant familial cerebral cavernous malformations. Here we report that native KRIT1 protein binds the effector loop of Rap1A but not H-Ras in a GTP-dependent manner, establishing that it is an authentic Rap1-specific effector. By modeling the KRIT1-Rap1 interface we designed a well-folded KRIT1 mutant that exhibited a ~40-fold-reduced affinity for Rap1A and maintained other KRIT1-binding functions. Direct binding of KRIT1 to Rap1 stabilized endothelial cell-cell junctions in vitro and was required for cardiovascular development in vivo. Mechanistically, Rap1 binding released KRIT1 from microtubules, enabling it to locate to cell--cell junctions, where it suppressed Rho kinase signaling and stabilized the junctions. These studies establish that the direct physical interaction of Rap1 with KRIT1 enables the translocation of microtubule-sequestered KRIT1 to junctions, thereby supporting junctional integrity and cardiovascular development.

journal_name

Mol Biol Cell

authors

Liu JJ,Stockton RA,Gingras AR,Ablooglu AJ,Han J,Bobkov AA,Ginsberg MH

doi

10.1091/mbc.E11-02-0157

subject

Has Abstract

pub_date

2011-07-15 00:00:00

pages

2509-19

issue

14

eissn

1059-1524

issn

1939-4586

pii

mbc.E11-02-0157

journal_volume

22

pub_type

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