Konno Akihiro, Kirby Martha, Anderson Stacie A, Schwartzberg Pamela L, Candotti Fabio
Genetics and Molecular Biology Branch, National Human Genome Research Institute, National Institutes of Health, 49 Convent Drive, Bethesday, MD 20892, USA.
Int Immunol. 2007 Feb;19(2):185-92. doi: 10.1093/intimm/dxl135. Epub 2007 Jan 6.
The Wiskott-Aldrich syndrome protein (WASP) is a key molecule for transduction of extracellular signals that induce a variety of critical biological events involving actin cytoskeleton rearrangement. Among the cellular partners of WASP, the Wiskott-Aldrich syndrome protein-interacting protein (WIP) has been speculated to play a critical role in the pathophysiology of Wiskott-Aldrich syndrome since WASP mutation hot spots map to the WIP-binding region. The notion that WIP promotes WASP function, however, conflicts with evidence that WIP inhibits WASP-mediated actin polymerization and IL-2 production and suggests a complex regulation of WASP function by WIP. Here we show that WASP gene transfer results in high WASP expression only when WIP is concomitantly expressed in K562 cells. Furthermore, WIP-knockdown experiments demonstrated that T cells with reduced WIP expression show a concordant reduction of WASP levels. Mapping studies using WIP mutants showed that the minimal WIP region able to rescue WASP expression in WIP-knockdown cells was the WASP-binding domain. However, expression of such a minimal domain of WIP failed to rescue WASP-dependent, nuclear factor of activated T-cells-mediated IL-2 transcriptional activity. These results demonstrate that expression of WIP is necessary for functional WASP expression in human cells and provide a new paradigm for understanding the function of these two molecules.
威斯科特-奥尔德里奇综合征蛋白(WASP)是转导细胞外信号的关键分子,这些信号可诱导多种涉及肌动蛋白细胞骨架重排的关键生物学事件。在WASP的细胞伴侣中,威斯科特-奥尔德里奇综合征蛋白相互作用蛋白(WIP)被推测在威斯科特-奥尔德里奇综合征的病理生理学中起关键作用,因为WASP突变热点映射到WIP结合区域。然而,WIP促进WASP功能的观点与WIP抑制WASP介导的肌动蛋白聚合和白细胞介素-2产生的证据相矛盾,这表明WIP对WASP功能有复杂的调节作用。在这里,我们表明,只有当WIP在K562细胞中同时表达时,WASP基因转移才会导致WASP高表达。此外,WIP敲低实验表明,WIP表达降低的T细胞中WASP水平也相应降低。使用WIP突变体的定位研究表明,能够挽救WIP敲低细胞中WASP表达的最小WIP区域是WASP结合域。然而,这种最小的WIP结构域的表达未能挽救WASP依赖性、活化T细胞核因子介导的白细胞介素-2转录活性。这些结果表明,WIP的表达对于人类细胞中功能性WASP的表达是必要的,并为理解这两种分子的功能提供了一个新的范例。