CHARACTERIZING A ROLE FOR NOS1AP IN CELLULAR MECHANOTRANSDUCTION

Abstract

In a screen for proteins that interact with the phosphotyrosine binding (PTB) domain of NOS1AP we identified the b-integrin family of proteins. Fibronectin stimulated mouse embryonic fibroblasts (MEFs) from NOS1AP-/- mice showed a significant increase in nuclear blebbing compared to controls. This defect could be rescued with the addition of the Rho Kinase inhibitor Y27632. Further characterization of MEFs lacking NOS1AP revealed a decrease in E-cadherin expression and an increase in N-cadherin expression. To further dissection of the role for NOS1AP in integrin dependent signalling, we turned to the RAW264.7 osteoclast-like cell line. He we found differentiated osteoclast-like cells showed an increase in NOS1AP expression. Using NOS1AP isoform specific antibodies revealed differential subcellular localization in differentiated osteoclast-like cells. Taken together, our data demonstrates that NOS1AP functions in integrin dependent signalling and that the different NOS1AP isoforms may contribute to the differentiation of osteoclasts.