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Regulation of sarcoma cell migration, invasion and invadopodia formation by AFAP1L1 through a phosphotyrosine-dependent pathway

Oncogene volume 35, pages 2098–2111 (2016)Cite this article

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Abstract

Invasion and metastasis are controlled by the invadopodia, which delivers matrix-degrading enzymes to the invasion interface permitting cancer cell penetration and spread into healthy tissue. We have identified a novel pathway that directs Lyn/Src family tyrosine kinase signals to the invadopodia to regulate sarcoma cell invasion via the molecule AFAP-1-like-1 (AFAP1L1), a new member of the AFAP (actin filament-associated protein) family. We show that AFAP1L1 can transform cells, promote migration and co-expression with active Lyn profoundly influences cell morphology and movement. AFAP1L1 intersects several invadopodia pathway components through its multiple domains and motifs, including the following (i) pleckstrin homology domains that bind phospholipids generated at the plasma membrane by phosphoinositide 3-kinase, (ii) a direct filamentous-actin binding domain and (iii) phospho-tyrosine motifs (pY136 and pY566) that specifically bind Vav2 and Nck2 SH2 domains, respectively. These phosphotyrosine motifs are essential for AFAP1L1-mediated cytoskeleton regulation. Through its interaction with Vav2, AFAP1L1 regulates Rac activity and downstream control of PAK1/2/3 (p21-activated kinases) phosphorylation of myosin light chain (MLC) kinase and MLC2. AFAP1L1 interaction with Nck2 recruits actin-nucleating complexes. Significantly, in osteosarcoma cell lines, knockdown of AFAP1L1 inhibits phosphorylated MLC2 recruitment to filamentous-actin structures, disrupts invadopodia formation, cell attachment, migration and invasion. These data define a novel pathway that directs Lyn/Src family tyrosine kinase signals to sarcoma cell invadopodia through specific recruitment of Vav2 and Nck2 to phosphorylated AFAP1L1, to control cell migration and invasion.

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Abbreviations

SFK:

Src family kinase

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Acknowledgements

We thank Janice Lam, Matt Lee, Rebecca Shapiro, Morgane Davies, Irma Larma and Kevin Li for technical assistance. This work was supported by grants from the National Health and Medical Research Council (513714 and 634352), the Medical Research Foundation of Royal Perth Hospital and the Cancer Council of Western Australia. EI received support from the Cancer Council of Western Australia, The Harry Perkins Institute of Medical Research, Sock-it-to-Sarcoma and the Hollywood Private Hospital Research Foundation.

Author contributions

DJMcC and SRT contributed equally to the manuscript and designed, supported and performed experiments, and analyzed data. TSK, AL, CL, JS and NK designed and performed experiments, and analyzed data. MP designed and analyzed experiments, and contributed to writing the manuscript. EI designed and supported the research, designed and undertook experiments, analyzed data and contributed to writing the manuscript.

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  1. S R Tie and D J McCarthy: These authors contributed equally to this work.

Authors and Affiliations

  1. Cell Signalling Group, Harry Perkins Institute of Medical Research and Centre for Medical Research, University of Western Australia, Nedlands, Western Australia, Australia

    S R Tie, D J McCarthy, T S Kendrick, A Louw, C Le, J Satiaputra, N Kucera & E Ingley

  2. Royal Perth Hospital Medical Research Foundation, Royal Perth Hospital, Perth, Western Australia, Australia

    A Louw

  3. Harry Perkins Institute of Medical Research and Centre for Medical Research, University of Western Australia, Perth, Western Australia, Australia

    M Phillips

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Tie, S., McCarthy, D., Kendrick, T. et al. Regulation of sarcoma cell migration, invasion and invadopodia formation by AFAP1L1 through a phosphotyrosine-dependent pathway. Oncogene 35, 2098–2111 (2016). https://doi.org/10.1038/onc.2015.272

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