Journal of Cell Science, Vol 109, Issue 4 817-826, Copyright © 1996 by Company of Biologists

JOURNAL ARTICLES

The relative roles of specific N- and C-terminal phosphorylation sites in the disassembly of intermediate filament in mitotic BHK-21 cells

YH Chou, P Opal, RA Quinlan and RD Goldman
Department of Cell and Molecular Biology, Northwestern University Medical School, Chicago, IL 60611, USA.

Previously we identified p34cdc2 as one of two protein kinases mediating the hyperphosphorylation and disassembly of vimentin in mitotic BHK-21 cells. In this paper, we identify the second kinase as a 37 kDa protein. This p37 protein kinase phosphorylates vimentin on two adjacent residues (thr-457 and ser-458) which are located in the C-terminal non-alpha-helical domain. Contrary to the p34cdc2 mediated N-terminal phosphorylation (at ser-55) which can disassemble vimentin intermediate filaments (IF) in vitro, p37 protein kinase phosphorylates vimentin-IF without obviously affecting its structure in vitro. We have further examined the in vivo role(s) of vimentin phosphorylation in the disassembly of the IF network in mitotic BHK cells by transient transfection assays. In untransfected BHK cells, the interphase vimentin IF networks are disassembled into non-filamentous aggregates when cells enter mitosis. Transfection of cells with vimentin cDNA lacking the p34cdc2 phosphorylation site (ser55:ala) effectively prevents mitotic cells from disassembling their IF. In contrast, apparently normal disassembly takes place in cells transfected with cDNA containing mutated p37 kinase phosphorylation sites (thr457:ala/ser458:ala). Transfection of cells with vimentin cDNAs lacking both the N- and C-terminal phosphorylation sites yields a phenotype indistinguishable from that obtained with the single N-terminal mutant. Taken together, our results demonstrate that the site-specific phosphorylation of the N-terminal domain, but not the C-terminal domain of vimentin plays an important role in determining the state of IF polymerization and supramolecular organization in mitotic cells.
CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    Twitter    What's this?

This article has been cited by other articles:

				Y.-H. Chou, W.-L. Kuo, M. R. Rosner, W.-J. Tang, and R. D. Goldman Structural changes in intermediate filament networks alter the activity of insulin-degrading enzyme FASEB J, November 1, 2009; 23(11): 3734 - 3742. [Abstract] [Full Text] [PDF]

				A. Bocquet, R. Berges, R. Frank, P. Robert, A. C. Peterson, and J. Eyer Neurofilaments Bind Tubulin and Modulate Its Polymerization J. Neurosci., September 2, 2009; 29(35): 11043 - 11054. [Abstract] [Full Text] [PDF]

				A. Zamoner, K. P. Barreto, D. W. Filho, F. Sell, V. M. Woehl, F. C. R. Guma, R. Pessoa-Pureur, and F. R. M. B. Silva Propylthiouracil-induced congenital hypothyroidism upregulates vimentin phosphorylation and depletes antioxidant defenses in immature rat testis J. Mol. Endocrinol., March 1, 2008; 40(3): 125 - 135. [Abstract] [Full Text] [PDF]

				S. Woll, R. Windoffer, and R. E. Leube p38 MAPK-dependent shaping of the keratin cytoskeleton in cultured cells J. Cell Biol., June 21, 2007; 177(5): 795 - 807. [Abstract] [Full Text] [PDF]

				E. R. Kuczmarski and R. D. Goldman Purification of Bovine Lens and Bacterially Expressed Human Vimentin Cold Spring Harb Protoc, February 1, 2007; 2007(3): pdb.prot4675 - pdb.prot4675. [Abstract] [Full Text]

				S. Ermakova, B. Y. Choi, H. S. Choi, B. S. Kang, A. M. Bode, and Z. Dong The Intermediate Filament Protein Vimentin Is a New Target for Epigallocatechin Gallate J. Biol. Chem., April 29, 2005; 280(17): 16882 - 16890. [Abstract] [Full Text] [PDF]

				J. E. Eriksson, T. He, A. V. Trejo-Skalli, A.-S. Harmala-Brasken, J. Hellman, Y.-H. Chou, and R. D. Goldman Specific in vivo phosphorylation sites determine the assembly dynamics of vimentin intermediate filaments J. Cell Sci., February 22, 2004; 117(6): 919 - 932. [Abstract] [Full Text] [PDF]

				Y.-H. Chou, S. Khuon, H. Herrmann, and R. D. Goldman Nestin Promotes the Phosphorylation-dependent Disassembly of Vimentin Intermediate Filaments During Mitosis Mol. Biol. Cell, April 1, 2003; 14(4): 1468 - 1478. [Abstract] [Full Text] [PDF]

				M. Al-Maghrebi, H. Brule, M. Padkina, C. Allen, W. M. Holmes, and Z. E. Zehner The 3' untranslated region of human vimentin mRNA interacts with protein complexes containing eEF-1{gamma} and HAX-1 Nucleic Acids Res., December 1, 2002; 30(23): 5017 - 5028. [Abstract] [Full Text] [PDF]

				K. H. Yoon, M. Yoon, R. D. Moir, S. Khuon, F. W. Flitney, and R. D. Goldman Insights into the Dynamic Properties of Keratin Intermediate Filaments in Living Epithelial Cells J. Cell Biol., April 30, 2001; 153(3): 503 - 516. [Abstract] [Full Text] [PDF]

				Y.-H. Chou and R. D. Goldman Intermediate Filaments on the Move J. Cell Biol., August 7, 2000; 150(3): F101 - F106. [Full Text] [PDF]

				Y. Yasui, M. Amano, K.-i. Nagata, N. Inagaki, H. Nakamura, H. Saya, K. Kaibuchi, and M. Inagaki Roles of Rho-associated Kinase in Cytokinesis; Mutations in Rho-associated Kinase Phosphorylation Sites Impair Cytokinetic Segregation of Glial Filaments J. Cell Biol., November 30, 1998; 143(5): 1249 - 1258. [Abstract] [Full Text] [PDF]

				W.-C. Sin, X.-Q. Chen, T. Leung, and L. Lim RhoA-Binding Kinase alpha  Translocation Is Facilitated by the Collapse of the Vimentin Intermediate Filament Network Mol. Cell. Biol., November 1, 1998; 18(11): 6325 - 6339. [Abstract] [Full Text] [PDF]

				M. Yoon, R. D. Moir, V. Prahlad, and R. D. Goldman Motile Properties of Vimentin Intermediate Filament Networks in Living Cells J. Cell Biol., October 5, 1998; 143(1): 147 - 157. [Abstract] [Full Text] [PDF]

				V. Prahlad, M. Yoon, R. D. Moir, R. D. Vale, and R. D. Goldman Rapid Movements of Vimentin on Microtubule Tracks: Kinesin-dependent Assembly of Intermediate Filament Networks J. Cell Biol., October 5, 1998; 143(1): 159 - 170. [Abstract] [Full Text] [PDF]

				S. Clement, P. T. Velasco, S. N. P. Murthy, J. H. Wilson, T. J. Lukas, R. D. Goldman, and L. Lorand The Intermediate Filament Protein, Vimentin, in the Lens Is a Target for Cross-linking by Transglutaminase J. Biol. Chem., March 27, 1998; 273(13): 7604 - 7609. [Abstract] [Full Text] [PDF]

				P. Vilalta, L Zhang, and S. Hamm-Alvarez A novel taxol-induced vimentin phosphorylation and stabilization revealed by studies on stable microtubules and vimentin intermediate filaments J. Cell Sci., January 7, 1998; 111(13): 1841 - 1852. [Abstract] [PDF]

				C. M. Sahlgren, A. Mikhailov, J. Hellman, Y.-H. Chou, U. Lendahl, R. D. Goldman, and J. E. Eriksson Mitotic Reorganization of the Intermediate Filament Protein Nestin Involves Phosphorylation by cdc2 Kinase J. Biol. Chem., May 4, 2001; 276(19): 16456 - 16463. [Abstract] [Full Text] [PDF]