QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

Vol. 15, Issue 3, 990-1002, March 2004

This Article Full Text Full Text (PDF) Supplemental Material All Versions of this Article: E03-09-0687v1 15/3/990    most recent Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Werner, N. S. Articles by Magin, T. M. Search for Related Content PubMed PubMed Citation Articles by Werner, N. S. Articles by Magin, T. M.

Epidermolysis Bullosa Simplex-Type Mutations Alter the Dynamics of the Keratin Cytoskeleton and Reveal a Contribution of Actin to the Transport of Keratin Subunits

Nicola Susann Werner ^*, Reinhard Windoffer , Pavel Strnad , Christine Grund , Rudolf Eberhard Leube , and Thomas Michael Magin ^* 

^* Institut fuer Physiologische Chemie, Abteilung fuer Zellbiochemie, Universitaetsklinikum Bonn, 53115 Bonn, Germany; Division of Cell Biology, German Cancer Research Center, 69120 Heidelberg, Germany; and Department of Anatomy, Johannes Gutenberg-University, D-55128 Mainz, Germany

Submitted September 24, 2003; Revised October 20, 2003; Accepted October 31, 2003
Monitoring Editor: Richard Hynes

Dominant keratin mutations cause epidermolysis bullosa simplex by transforming keratin (K) filaments into aggregates. As a first step toward understanding the properties of mutant keratins in vivo, we stably transfected epithelial cells with an enhanced yellow fluorescent protein-tagged K14R₁₂₅C mutant. K14R₁₂₅C became localized as aggregates in the cell periphery and incorporated into perinuclear keratin filaments. Unexpectedly, keratin aggregates were in dynamic equilibrium with soluble subunits at a half-life time of <15 min, whereas filaments were extremely static. Therefore, this dominant-negative mutation acts by altering cytoskeletal dynamics and solubility. Unlike previously postulated, the dominance of mutations is limited and strictly depends on the ratio of mutant to wild-type protein. In support, K14R₁₂₅C-specific RNA interference experiments resulted in a rapid disintegration of aggregates and restored normal filaments. Most importantly, live cell inhibitor studies revealed that the granules are transported from the cell periphery inwards in an actin-, but not microtubule-based manner. The peripheral granule zone may define a region in which keratin precursors are incorporated into existing filaments. Collectively, our data have uncovered the transient nature of keratin aggregates in cells and offer a rationale for the treatment of epidermolysis bullosa simplex by using short interfering RNAs.

Abbreviations used: EBS, epidermolysis bullosa simplex; EYFP, enhanced yellow fluorescent protein; IF, intermediate filament; K, keratin; MF, microfilament; MT, microtubule; mut, mutant; wt, wild type.

Online version of this article contains video and supplementary material for some figures. Online version is available at www.molbiolcell.org.

Corresponding author. E-mail address: t.magin{at}uni-bonn.de.

This article has been cited by other articles:

				S. Kim and P. A. Coulombe Intermediate filament scaffolds fulfill mechanical, organizational, and signaling functions in the cytoplasm Genes & Dev., July 1, 2007; 21(13): 1581 - 1597. [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]

				H. A. Long, V. Boczonadi, L. McInroy, M. Goldberg, and A. Maatta Periplakin-dependent re-organisation of keratin cytoskeleton and loss of collective migration in keratin-8-downregulated epithelial sheets J. Cell Sci., December 15, 2006; 119(24): 5147 - 5159. [Abstract] [Full Text] [PDF]

				R. P HICKERSON, F. J. D SMITH, W. H IRWIN MCLEAN, M. LANDTHALER, R. E LEUBE, and R. L KASPAR SiRNA-Mediated Selective Inhibition of Mutant Keratin mRNAs Responsible for the Skin Disorder Pachyonychia Congenita Ann. N.Y. Acad. Sci., October 1, 2006; 1082(1): 56 - 61. [Abstract] [Full Text] [PDF]

				S. Osmanagic-Myers, M. Gregor, G. Walko, G. Burgstaller, S. Reipert, and G. Wiche Plectin-controlled keratin cytoarchitecture affects MAP kinases involved in cellular stress response and migration J. Cell Biol., August 14, 2006; 174(4): 557 - 568. [Abstract] [Full Text] [PDF]

				R. Windoffer, A. Kolsch, S. Woll, and R. E. Leube Focal adhesions are hotspots for keratin filament precursor formation J. Cell Biol., May 8, 2006; 173(3): 341 - 348. [Abstract] [Full Text] [PDF]

				M. Nishizawa, I. Izawa, A. Inoko, Y. Hayashi, K.-i. Nagata, T. Yokoyama, J. Usukura, and M. Inagaki Identification of trichoplein, a novel keratin filament-binding protein J. Cell Sci., March 1, 2005; 118(5): 1081 - 1090. [Abstract] [Full Text] [PDF]

				L.-H. Gu and P. A. Coulombe Defining the Properties of the Nonhelical Tail Domain in Type II Keratin 5: Insight from a Bullous Disease-causing Mutation Mol. Biol. Cell, March 1, 2005; 16(3): 1427 - 1438. [Abstract] [Full Text] [PDF]

				M. B. Omary, P. A. Coulombe, and W.H. I. McLean Intermediate Filament Proteins and Their Associated Diseases N. Engl. J. Med., November 11, 2004; 351(20): 2087 - 2100. [Full Text] [PDF]