The role of vimentin in the tumor marker Nup88-dependent multinucleated phenotype

Abstract

BackgroundNucleoporin Nup88, a component of nuclear pore complexes, is known to be overexpressed in several types of tumor tissue. The overexpression of Nup88 has been reported to promote the early step of tumorigenesis by inducing multinuclei in both HeLa cells and a mouse model. However, the molecular basis of how Nup88 leads to a multinucleated phenotype remains unclear because of a lack of information concerning its binding partners. In this study, we characterize a novel interaction between Nup88 and vimentin. We also examine the involvement of vimentin in the Nup88-dependent multinucleated phenotype.MethodsCells overexpressing tagged versions of Nup88, vimentin and their truncations were used in this study. Coprecipitation and GST-pulldown assays were carried out to analyze protein-protein interactions. Vimentin knockdown by siRNA was performed to examine the functional role of the Nup88-vimentin interaction in cells. The phosphorylation status of vimentin was analyzed by immunoblotting using an antibody specific for its phosphorylation site.ResultsVimentin was identified as a Nup88 interacting partner, although it did not bind to other nucleoporins, such as Nup50, Nup214, and Nup358, in HeLa cell lysates. The N-terminal 541 amino acid residues of Nup88 was found to be responsible for its interaction with vimentin. Recombinant GST-tagged Nup88 bound to recombinant vimentin in a GST-pulldown assay. Although overexpression of Nup88 in HeLa cells was observed mainly at the nuclear rim and in the cytoplasm, colocalization with vimentin was only partially detected at or around the nuclear rim. Disruption of the Nup88-vimentin interaction by vimentin specific siRNA transfection suppressed the Nup88-dependent multinucleated phenotype. An excess amount of Nup88 in cell lysates inhibited the dephosphorylation of a serine residue (Ser83) within the vimentin N-terminal region even in the absence and presence of an exogenous phosphatase. The N-terminal 96 amino acid residues of vimentin interacted with both full-length and the N-terminal 541 residues of Nup88.ConclusionsNup88 can affect the phosphorylation status of vimentin, which may contribute to the Nup88-dependent multinucleated phenotype through changing the organization of vimentin.

A flexible network of vimentin intermediate filaments promotes migration of amoeboid cancer cells through confined environments

“…Consistent with previous reports, our measurements of stiffness found that Vimentin protects the cell against compression (29). Vimentin may be particularly important for protecting the nucleus from compressive force, as several adaptor proteins have been reported to connect it to the nuclear envelope (30)(31)(32). Accordingly, Vimentin has been recently reported to protect the nucleus from rupture and DNA damage in confinement (33,34).…”

A flexible network of Vimentin intermediate filaments promotes the migration of amoeboid cancer cells through confined environments

“…Consistent with previous reports, our measurements of stiffness found that Vimentin protects the cell against compression (24). Vimentin may be particularly important for protecting the nucleus from compressive force, as several adaptor proteins have been reported to connect it to the nuclear envelope (25)(26)(27). Accordingly, Vimentin has been recently reported to protect the nucleus from rupture and DNA damage in confinement (28,29).…”

Long non‐coding RNA MILNR1 retards colorectal cancer growth by inhibiting c‐Myc

“…Nucleoporin 88 (NUP88) is a component of nucleoporins, which is upregulated in tumor tissues including CRC [8]. It is reported that NUP88 interacts with vimentin and protects its serine residue (Ser83) from dephosphorylation, thereby promoting cell proliferation [9]. Another form of vimentin phosphorylation, namely the phosphorylation at Ser39, results in the development of cell migration and tumor metastasis [10].…”