title: Human enhancer of filamentation 1-induced colorectal cancer cell migration: Role of serine phosphorylation and interaction with the breast cancer anti-estrogen resistance 3 protein.
abstract: Human enhancer of filamentation 1 (HEF1) is a member of the p130Cas family of docking proteins involved in integrin-mediated cytoskeleton reorganization associated with cell migration. Elevated expression of HEF1 promotes invasion and metastasis in multiple cancer cell types. To date, little is known on its role in CRC tumor progression. HEF1 is phosphorylated on several Ser/Thr residues but the effects of these post-translational modifications on the functions of HEF1 are poorly understood. In this manuscript, we investigated the role of HEF1 in migration of colorectal adeno-carcinoma cells. First, we showed that overexpression of HEF1 in colo-carcinoma cell line HCT116 increases cell migration. Moreover, in these cells, HEF1 increases Src-mediated phosphorylation of FAK on Tyr-861 and 925. We then showed that HEF1 mutation on Ser-369 enhances HEF1-induced migration and FAK phosphorylation as a result of protein stabilization. We also, for the first time characterized a functional mutation of HEF1 on Arg-367 which mimics the effect of Ser-369 to Ala mutation. Finally through mass spectrometry experiments, we identified BCAR3 as an essential interactor and mediator of HEF1-induced migration. We demonstrated that single amino acid mutations that prevent formation of the HEF1-BCAR3 complex impair HEF1-mediated migration. Therefore, amino-acid substitutions that impede Ser-369 phosphorylation stabilize HEF1 which increases the migration of CRC cells and this latter effect requires the interaction of HEF1 with the NSP family adaptor protein BCAR3. Collectively, these data reveal the importance of HEF1 expression level in cancer cell motility and then support the utilization of HEF1 as a biomarker of tumor progression.
authors: INSERM U749, Institut Gustave Roussy, Université Paris-sud, Villejuif 94800, France; Department of Biochemistry and Microbiology, Faculty of Pharmacy, Tichrine University, Latakia, Syria.
authors: Rama
authors: R
authors: Ibrahim
authors: INSERM U1004, Laboratoire de biochimie, Hopital Paul Brousse, Université Paris-sud, Villejuif 94800, France.
authors: Antoinette
authors: A
authors: Lemoine
authors: INSERM U749, Institut Gustave Roussy, Université Paris-sud, Villejuif 94800, France.
authors: Jacques
authors: J
authors: Bertoglio
authors: INSERM U749, Institut Gustave Roussy, Université Paris-sud, Villejuif 94800, France. Electronic address: joel.raingeaud@gustaveroussy.fr.
authors: Joël
authors: J
authors: Raingeaud
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000502:physiology*; D002465:Cell Movement; D020662:Guanine Nucleotide Exchange Factors; D045325:HCT116 Cells; D006801:Humans; D010750:Phosphoproteins / Q000502:physiology*; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational
chemical_list: D048868:Adaptor Proteins, Signal Transducing; C118588:BCAR3 protein, human; D020662:Guanine Nucleotide Exchange Factors; C099942:NEDD9 protein, human; D010750:Phosphoproteins
keywords: BCAR3; Colorectal cancer; Focal adhesion kinase; Nedd9; Phosphorylation