Role Of Nucleoporins In Modulating Differentiation In Embryonal Carcinoma Cells

Abstract

NTERA-2 cl.D1 (NT2/D1) is an embryonal carcinoma cell line that differentiates into a neuronal phenotype upon induction with retinoic acid (RA). Cellular differentiation incorporates various stimuli and involves multiple layers of regulation in determining cell fate. In NT2/D1 cells, the nucleoporin Nup93, regulates the expression of the HOXA gene cluster during differentiation. Nucleoporins are channel proteins embedded in the nuclear envelope and are canonically involved in nucleocytoplasmic transport of RNA and proteins. However, recent studies implicate their roles in gene regulation and cell fate determination. Here we investigated the role of the Nup93-subcomplex proteins - Nup93, Nup188 and Nup205 in differentiation. We show that Nup93 subcomplex proteins regulate the expression of pluripotency and lineage genes. Furthermore, induction of differentiation upon RA treatment reveals a differential role of Nup93 and Nup188 on differentiation. Here we show a novel role for Nup205 in the formation of cell aggregates in embryonal carcinoma cells. We also showed that RA treatment in NT2/D1 differentially altered lamin protein levels, and examined lamin function during differentiation. In addition, preliminary data substantiates the contribution of mechanosignaling in differentiation. Taken together our data reveals a regulatory role for the Nup93 subcomplex in modulating neuronal differentiation in NT2/D1 cells.