Tyrosine phosphorylation of WIP releases bound WASP and impairs podosome assembly in macrophages
Tyrosine phosphorylation of WIP releases bound WASP and impairs podosome assembly in macrophages
复制标题
WIP 的酪氨酸磷酸化释放结合的 WASP 并损害巨噬细胞中的足小体组装
作者：
V. Vijayakumar;J. Monypenny;X. Chen;L. Machesky;S. Lilla;A. Thrasher;I. Antón;Y. Calle;G. Jones
摘要 足体是含有整合素的粘附结构，常见于单核细胞谱系的迁移白细胞中。足体的肌动蛋白细胞骨架组织基于 WASP 和 Arp2/3 介导的机制。 WASP 还与第二种蛋白质 WIP（也称为 WIPF1）相关，并且它们共定位于足小体核心中。在这里，我们首次报道 WIP 可以在酪氨酸残基上磷酸化，并且 WIP 的酪氨酸磷酸化是 WIP-WASP 复合物中释放 WASP 的触发因素。通过使用敲除方法和 WIP 磷酸模拟物的表达，我们发现在没有 WIP-WASP 结合的情况下，细胞 WASP 会迅速降解，导致足体破坏和细胞无法降解底层基质。在没有酪氨酸磷酸化的情况下，WIP-WASP 复合体保持完整，足体寿命延长。候选激酶的筛选和基于抑制剂的测定确定了布鲁顿酪氨酸激酶 (Btk) 作为 WIP 酪氨酸磷酸化的调节剂。我们得出结论，WIP 的酪氨酸磷酸化是 WASP 稳定性的关键调节因子，并且具有肌动蛋白成核促进因子的功能。
ABSTRACT Podosomes are integrin-containing adhesion structures commonly found in migrating leukocytes of the monocytic lineage. The actin cytoskeletal organisation of podosomes is based on a WASP- and Arp2/3-mediated mechanism. WASP also associates with a second protein, WIP (also known as WIPF1), and they co-localise in podosome cores. Here, we report for the first time that WIP can be phosphorylated on tyrosine residues and that tyrosine phosphorylation of WIP is a trigger for release of WASP from the WIP–WASP complex. Using a knockdown approach together with expression of WIP phosphomimics, we show that in the absence of WIP–WASP binding, cellular WASP is rapidly degraded, leading to disruption of podosomes and a failure of cells to degrade an underlying matrix. In the absence of tyrosine phosphorylation, the WIP–WASP complex remains intact and podosome lifetimes are extended. A screen of candidate kinases and inhibitor-based assays identified Bruton's tyrosine kinase (Btk) as a regulator of WIP tyrosine phosphorylation. We conclude that tyrosine phosphorylation of WIP is a crucial regulator of WASP stability and function as an actin-nucleation-promoting factor.
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影响因子：
1.6
作者：
Wu B;Wang Z;Lin N;Yan X;Lv Z;Ying Z;Ye Z
通讯作者：
Ye Z
DOI：
10.1073/pnas.94.26.14671
发表时间：
1997-12-23
影响因子：
11.1
作者：
Ramesh, N;Ant贸n, IM;Geha, RS
通讯作者：
Geha, RS
影响因子：
20.3
作者：
Tai, Yu-Tzu;Chang, Betty Y.;Anderson, Kenneth C.
通讯作者：
Anderson, Kenneth C.
影响因子：
4
作者：
Brandvold, Kristoffer R.;Steffey, Michael E.;Fox, Christel C.;Soellner, Matthew B.
通讯作者：
Soellner, Matthew B.
影响因子：
20.3
作者：
Cougoule, Celine;Le Cabec, Veronique;Maridonneau-Parini, Isabelle
通讯作者：
Maridonneau-Parini, Isabelle