Khurana Bharat, Khurana Taruna, Khaire Nandkumar, Noegel Angelika A
Center for Biochemistry, Medical Faculty, University of Cologne, Joseph-Stelzmann-Strasse 52, D-50931 Cologne, Germany.
EMBO J. 2002 Oct 15;21(20):5331-42. doi: 10.1093/emboj/cdf550.
LimC and LimD are two novel LIM proteins of Dictyostelium, which are comprised of double and single LIM domains, respectively. Green fluorescent protein-fused LimC and LimD proteins preferentially accumulate at areas of the cell cortex where they co-localize with actin and associate transiently with cytoskeleton-dependent dynamic structures like phagosomes, macropinosomes and pseudopods. Furthermore, both LimC and LimD interact directly with F-actin in vitro. Mutant cells that lack either LimC or LimD, or both, exhibit normal growth. They are, however, significantly impaired in growth under stress conditions and are highly sensitive to osmotic shock, suggesting that LimC and LimD contribute towards the maintenance of cortical strength. Moreover, we noted an altered morphology and F-actin distribution in LimD(-) and LimC(-)/D(-) mutants, and changes in chemotactic motility associated with an increased pseudopod formation. Our results reveal both unique and overlapping roles for LimC and LimD, and suggest that both act directly on the actin cytoskeleton and provide rigidity to the cortex.
LimC和LimD是盘基网柄菌的两种新型LIM蛋白,分别由双LIM结构域和单LIM结构域组成。绿色荧光蛋白融合的LimC和LimD蛋白优先聚集在细胞皮层区域,在那里它们与肌动蛋白共定位,并与吞噬体、巨胞饮体和伪足等细胞骨架依赖性动态结构短暂结合。此外,LimC和LimD在体外都能直接与F-肌动蛋白相互作用。缺乏LimC或LimD或两者都缺乏的突变细胞表现出正常生长。然而,它们在应激条件下的生长显著受损,并且对渗透压休克高度敏感,这表明LimC和LimD有助于维持皮层强度。此外,我们注意到LimD(-)和LimC(-)/D(-)突变体的形态和F-肌动蛋白分布发生了改变,并且趋化运动性的变化与伪足形成增加有关。我们的结果揭示了LimC和LimD独特和重叠的作用,并表明两者都直接作用于肌动蛋白细胞骨架并为皮层提供刚性。