title: Regulation of sarcoma cell migration, invasion and invadopodia formation by AFAP1L1 through a phosphotyrosine-dependent pathway.
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.
authors.affiliation: Cell Signalling Group, Harry Perkins Institute of Medical Research and Centre for Medical Research, University of Western Australia, Nedlands, Western Australia, Australia.
authors.forename: S R
authors.initials: SR
authors.lastname: Tie
authors.affiliation: Cell Signalling Group, Harry Perkins Institute of Medical Research and Centre for Medical Research, University of Western Australia, Nedlands, Western Australia, Australia.
authors.forename: D J
authors.initials: DJ
authors.lastname: McCarthy
authors.affiliation: Cell Signalling Group, Harry Perkins Institute of Medical Research and Centre for Medical Research, University of Western Australia, Nedlands, Western Australia, Australia.
authors.forename: T S
authors.initials: TS
authors.lastname: Kendrick
authors.affiliation: Cell Signalling Group, Harry Perkins Institute of Medical Research and Centre for Medical Research, University of Western Australia, Nedlands, Western Australia, Australia.|Royal Perth Hospital Medical Research Foundation, Royal Perth Hospital, Perth, Western Australia, Australia.
authors.forename: A
authors.initials: A
authors.lastname: Louw
authors.affiliation: Cell Signalling Group, Harry Perkins Institute of Medical Research and Centre for Medical Research, University of Western Australia, Nedlands, Western Australia, Australia.
authors.forename: C
authors.initials: C
authors.lastname: Le
authors.affiliation: Cell Signalling Group, Harry Perkins Institute of Medical Research and Centre for Medical Research, University of Western Australia, Nedlands, Western Australia, Australia.
authors.forename: J
authors.initials: J
authors.lastname: Satiaputra
authors.affiliation: Cell Signalling Group, Harry Perkins Institute of Medical Research and Centre for Medical Research, University of Western Australia, Nedlands, Western Australia, Australia.
authors.forename: N
authors.initials: N
authors.lastname: Kucera
authors.affiliation: Harry Perkins Institute of Medical Research and Centre for Medical Research, University of Western Australia, Perth, Western Australia, Australia.
authors.forename: M
authors.initials: M
authors.lastname: Phillips
authors.affiliation: Cell Signalling Group, Harry Perkins Institute of Medical Research and Centre for Medical Research, University of Western Australia, Nedlands, Western Australia, Australia.
authors.forename: E
authors.identifier: http://orcid.org/0000-0002-8112-9134
authors.initials: E
authors.lastname: Ingley
mesh_terms: D000199:Actins / Q000378:metabolism; D048868:Adaptor Proteins, Signal Transducing / Q000502:physiology*; D000818:Animals; D002455:Cell Division; D045744:Cell Line, Tumor; D006801:Humans; D051379:Mice; D008840:Microfilament Proteins / Q000502:physiology*; D009361:Neoplasm Invasiveness; D009362:Neoplasm Metastasis; D010766:Phosphorylation; D019000:Phosphotyrosine / Q000378:metabolism*; D012509:Sarcoma / Q000378:metabolism / Q000473:pathology*
chemical_list: C559217:AFAP1L1 protein, human; D000199:Actins; D048868:Adaptor Proteins, Signal Transducing; D008840:Microfilament Proteins; D019000:Phosphotyrosine