Epidermolysis Bullosa Simplex-Type Mutations Alter the Dynamics of ...

5 downloads 0 Views 2MB Size Report
Bar, 10 μm. (B) Projection of the first 30 min from Movie 1 into a single picture depicting the area of aggregate turn- over. (C) Time space diagram derived from a.
Molecular Biology of the Cell Vol. 15, 990 –1002, March 2004

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