Paper Citing NAMD - Abstract
Liu, Jian J.; Stockton, Rebecca A.; Gingras, Alexandre R.; Ablooglu, Ararat J.; Han, Jaewon; Bobkov, Andrey A.; Ginsberg, Mark H.
A mechanism of Rap1-induced stabilization of endothelial cell-cell junctions
MOLECULAR BIOLOGY OF THE CELL, 22:2509-2519, JUL 15 2011
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 similar to 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.
