Gryaznova Tetyana, Kropyvko Sergii, Burdyniuk Mariia, Gubar Olga, Kryklyva Valentyna, Tsyba Liudmyla, Rynditch Alla
Institute of Molecular Biology and Genetics, 150 Zabolotnogo Street, Kyiv 03680, Ukraine.
Institute of Molecular Biology and Genetics, 150 Zabolotnogo Street, Kyiv 03680, Ukraine.
Cell Signal. 2015 Jul;27(7):1499-508. doi: 10.1016/j.cellsig.2015.03.006. Epub 2015 Mar 19.
Invasive cancer cells form actin-rich membrane protrusions called invadopodia that degrade extracellular matrix and facilitate cell invasion and metastasis. WIP (WASP-interacting protein) together with N-WASP (neural Wiskott-Aldrich syndrome protein) are localized in invadopodia and play a crucial role in their formation. Here we show that WIP interacts with endocytic adaptor proteins of the intersectin (ITSN) family, ITSN1 and ITSN2. The interaction is mediated by the SH3 domains of ITSNs and the middle part of the WIP proline-rich motifs. We have also demonstrated that ITSN1, WIP and N-WASP can form a complex in cells. Endogenous ITSN1 and ITSN2 are located in invasive protrusions of MDA-MB-231 breast cancer cell line. Moreover, data from immunofluorescent analysis revealed co-localization of ITSN1 and WIP at sites of invadopodia formation and in clathrin-coated pits. Together, these findings provide insights into the molecular mechanisms of invadopodia formation and identify ITSNs as scaffold proteins involved in this process.
侵袭性癌细胞形成富含肌动蛋白的膜突出结构，称为侵袭伪足，其可降解细胞外基质并促进细胞侵袭和转移。WIP（WASP相互作用蛋白）与N-WASP（神经威斯科特-奥尔德里奇综合征蛋白）共同定位于侵袭伪足中，并在其形成过程中发挥关键作用。在此，我们表明WIP与相交蛋白（ITSN）家族的内吞衔接蛋白ITSN1和ITSN2相互作用。这种相互作用由ITSNs的SH3结构域和WIP富含脯氨酸基序的中间部分介导。我们还证明了ITSN1、WIP和N-WASP可在细胞中形成复合物。内源性ITSN1和ITSN2位于MDA-MB-231乳腺癌细胞系的侵袭性突出结构中。此外，免疫荧光分析数据显示ITSN1和WIP在侵袭伪足形成部位和网格蛋白包被小窝中共定位。这些发现共同为侵袭伪足形成的分子机制提供了见解，并将ITSNs鉴定为参与这一过程的支架蛋白。
