Nestin Promotes the Phosphorylation-dependent Disassembly of Vimentin Intermediate Filaments During Mitosis

Department of Cell and Molecular Biology, Northwestern University, 303 East Chicago Avenue, Chicago, IL 60611, USA.
Molecular Biology of the Cell (Impact Factor: 4.47). 05/2003; 14(4):1468-78. DOI: 10.1091/mbc.E02-08-0545
Source: PubMed


The expression of the intermediate filament (IF) protein nestin is closely associated with rapidly proliferating progenitor cells during neurogenesis and myogenesis, but little is known about its function. In this study, we examine the effects of nestin expression on the assembly state of vimentin IFs in nestin-free cells. Nestin is introduced by transient transfection and is positively correlated with the disassembly of vimentin IFs into nonfilamentous aggregates or particles in mitotic but not interphase cells. This nestin-mediated disassembly of IFs is dependent on the phosphorylation of vimentin by the maturation/M-phase-promoting factor at ser-55 in the amino-terminal head domain. In addition, the disassembly of vimentin IFs during mitosis appears to be a unique feature of nestin-expressing cell types. Furthermore, when the expression of nestin is downregulated by the nestin-specific small interfering RNA in nestin-expressing cells, vimentin IFs remain assembled throughout all stages of mitosis. Previous studies suggest that nonfilamentous vimentin particles are IF precursors and can be transported rapidly between different cytoplasmic compartments along microtubule tracks. On the basis of these observations, we speculate that nestin may play a role in the trafficking and distribution of IF proteins and potentially other cellular factors to daughter cells during progenitor cell division.

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Available from: Harald Herrmann, Jan 29, 2015
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    • "Nestin interacts with other IF proteins including vimentin, desmin, and synemin to form heterodimers and mixed polymers [9] [10] [11] [12]. Vimentin is a major partner for nestin, and nestin regulates the reorganization, disassembly , and phosphorylation of vimentin during mitosis [13]. Copolymers of nestin and vimentin serve as scaffolds for glucocorticoid receptor molecules [14]. "
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    ABSTRACT: Nestin, a class VI intermediate filament, was first described as a neuronal stem/progenitor cell marker. We previously reported that knockdown of nestin expression in human glioblastoma cells suppresses cell proliferation, migration, and invasion. In the present study, we examined the effect of nestin on stemness, and identified molecules involved in modulating nestin function in glioblastoma cells. Nestin expression was shown to be higher in high-grade gliomas than in low-grade gliomas. Furthermore, compared with control cells, nestin short hairpin RNA (shRNA)-transfected glioblastoma cells exhibited reduced sphere formation, decreased expression of NANOG, N-cadherin, CD133, and Oct-4, and decreased tumor size in vivo. To examine the proteins regulated by nestin in glioblastomas, we carried out two-dimensional electrophoresis using nestin shRNA-transfected glioblastoma cells. As a result, nestin shRNA-transfected glioblastoma cells exhibited a decrease in the level of phosphorylation of heat shock cognate 71 kDa protein (HSC71; gene HSPA8). From immunoprecipitation experiments, we demonstrated the direct binding of nestin, HSC71, and cyclin D1 in vitro. Overexpression of nestin in glioblastoma cells increased cell growth, sphere formation, and cell invasion. Transfection with HSC71 siRNA restored nestin expression and cell behavior; therefore, HSC71 knockdown will interfere with enhanced tumorigenic properties of glioblastoma cells that ectopically overexpress nestin. We have demonstrated that HSC71 and nestin regulate each other's expression levels or patterns, and that cyclin D1 is located downstream of nestin and HSC71. In conclusion, nestin regulates stemness, cell growth, and invasion in glioblastoma cells through the alteration of HSC71. Inhibition of nestin and HSC71 may thus be a useful molecular target in the treatment of glioblastomas. Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.
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    • "We cannot exclude that there are other factors which we did not investigate that could have caused the increase of soluble vimentin in DBP-treated Sertoli cells, but phosphorylation of Ser 83 is one of the contributing factors. Previous studies have showed that phosphorylation of the Nterminal non--helical domain of vimentin was involved in the assembly/disassembly of vimentin filaments545556. Residue 83 of vimentin was one of the phosphorylation sites which resulted in an increase in the distance between head domains. "
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    ABSTRACT: Dibutyl phthalate (DBP) is a peroxisome proliferator which can lead to germ cell loss from Sertoli cells. Collapse of vimentin filaments occur in Sertoli cells after DBP exposure. Peroxisome proliferator activated receptor α (PPARα) is a key receptor which could be activated by DBP. The role of PPARα in this process was investigated. Results showed that, PPARα was activated in DBP-exposed Sertoli cells, GW6471 inhibited the activity of PPARα, phosphorylation level of vimentin and concentration of soluble vimentin was higher in DBP-treated Sertoli cells than GW6471 + DBP-treated cells. These results suggest that PPARα directly or indirectly mediated phosphorylation of vimentin on Ser 83, and PPARα may play an important role in regulating the reorganization of vimentin filaments during exposure of Sertoli cells to DBP.
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    • "Recent studies have revealed vimentin as highly dynamic polymer, with three types of structures (granules, squiggles, and filaments) coexisting in the cell. Particularly vimentin IF varies in the form of granules and squiggles and have an important role in the assembly and disassembly of IF networks [51]. Vimentin is involved in many important physiological functions, such as signal transduction, gene regulation and distributions of organelles [52]. "
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    ABSTRACT: Clostridium botulinum C3 exoenzyme (C3) selectively inactivates RhoA/B/C GTPases by ADP-ribosylation. Based on this substrate specificity C3 is a well-established tool in cell biology. C3 is taken up by eukaryotic cells although lacking an uptake and translocation domain. Based on different approaches vimentin was identified as membranous C3-interaction partner by mass spectrometry. Vimentin in fact was partly localized at the outer surface of hippocampal HT22 cells and J744A.1 macrophages. Domain analysis identified the rod domain as binding partner of C3. Vimentin was also involved in uptake of C3 as shown by knock down of vimentin in HT22 and J774A.1 cells. The involvement of vimentin in uptake of C3 was further supported by the findings that the vimentin disruptor acrylamide blocked uptake of C3. Vimentin is not only a major organizing element of the intermediate filament network but is also involved in both binding and uptake of C3 exoenzyme.
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