Conway James R W, Joshi Omkar, Kaivola Jasmin, Follain Gautier, Gounis Michalis, Kühl David, Ivaska Johanna
Turku Bioscience Centre, University of Turku and Åbo Akademi University, Turku, Finland.
Department of Biochemistry and Developmental Biology, Faculty of Medicine, University of Helsinki, Helsinki, Finland.
Nat Cell Biol. 2025 May 26. doi: 10.1038/s41556-025-01663-4.
Integrins provide an essential bridge between cancer cells and the extracellular matrix, playing a central role in every stage of disease progression. Despite the recognized importance of integrin phosphorylation in several biological processes, the regulatory mechanisms and their relevance remained elusive. Here we engineer a fluorescence resonance energy transfer biosensor for integrin β1 phosphorylation, screening 96 protein tyrosine phosphatases and identifying Shp2 and PTP-PEST as negative regulators to address this gap. Mutation of the integrin NPxY(783/795) sites revealed the importance of integrin phosphorylation for efficient cancer cell invasion, further supported by inhibition of the identified integrin phosphorylation regulators Shp2 and Src kinase. Using proteomics approaches, we uncovered Cofilin as a component of the phosphorylated integrin-Dok1 complex and linked this axis to effective invadopodia formation, a process supporting breast cancer invasion. These data further implicate dynamic modulation of integrin β1 phosphorylation at NPxY sites at different stages of metastatic dissemination.
整合素在癌细胞与细胞外基质之间提供了至关重要的桥梁，在疾病进展的各个阶段都发挥着核心作用。尽管整合素磷酸化在多个生物学过程中的重要性已得到认可，但其调控机制及其相关性仍不清楚。在此，我们设计了一种用于整合素β1磷酸化的荧光共振能量转移生物传感器，筛选了96种蛋白质酪氨酸磷酸酶，并确定Shp2和PTP-PEST为负调控因子，以填补这一空白。整合素NPxY(783/795)位点的突变揭示了整合素磷酸化对癌细胞有效侵袭的重要性，对已鉴定的整合素磷酸化调节因子Shp2和Src激酶的抑制进一步支持了这一点。通过蛋白质组学方法，我们发现丝切蛋白是磷酸化整合素-Dok1复合物的一个组成部分，并将该轴与有效的侵袭伪足形成联系起来，侵袭伪足形成是支持乳腺癌侵袭的一个过程。这些数据进一步表明，在转移扩散的不同阶段，整合素β1在NPxY位点的磷酸化存在动态调节。
