Paracingulin recruits CAMSAP3 to tight junctions and regulates microtubule and polarized epithelial organization.

Paracingulin (CGNL1) is recruited to tight junctions (TJ) by ZO-1 and to adherens junctions (AJ) by PLEKHA7. PLEKHA7 was reported to bind to the microtubule minus-end binding protein CAMSAP3, to tether microtubules to the AJ. Here we show that the KO of CGNL1, but not of PLEKHA7, results in the loss of junctional CAMSAP3 and its redistribution to a cytoplasmic pool both in cultured epithelial cells in vitro and mouse intestinal epithelium in vivo. In agreement, GST pulldown analyses show that CGNL1 but not PLEKHA7 interacts strongly with CAMSAP3, and the interaction is mediated by their respective coiled-coil regions. Ultrastructure-expansion microscopy shows that CAMSAP3-capped microtubules are tethered to junctions by the ZO-1-associated pool of CGNL1. The KO of CGNL1 results in disorganized cytoplasmic microtubules and irregular nuclei alignment in mouse intestinal epithelial cells, altered cyst morphogenesis in cultured kidney epithelial cells, and disrupted planar apical microtubules in mammary epithelial cells. Together, these results uncover new functions of CGNL1 in recruiting CAMSAP3 to junctions and regulating microtubule cytoskeleton organization and epithelial cell architecture.