RESUMO
Tankyrase, a PARP that promotes telomere elongation and Wnt/ß-catenin signaling, has various binding partners, suggesting that it has as-yet unidentified functions. Here, we report that the tankyrase-binding protein TNKS1BP1 regulates actin cytoskeleton and cancer cell invasion, which is closely associated with cancer progression. TNKS1BP1 colocalized with actin filaments and negatively regulated cell invasion. In TNKS1BP1-depleted cells, actin filament dynamics, focal adhesion, and lamellipodia ruffling were increased with activation of the ROCK/LIMK/cofilin pathway. TNKS1BP1 bound the actin-capping protein CapZA2. TNKS1BP1 depletion dissociated CapZA2 from the cytoskeleton, leading to cofilin phosphorylation and enhanced cell invasion. Tankyrase overexpression increased cofilin phosphorylation, dissociated CapZA2 from cytoskeleton, and enhanced cell invasion in a PARP activity-dependent manner. In clinical samples of pancreatic cancer, TNKS1BP1 expression was reduced in invasive regions. We propose that the tankyrase-TNKS1BP1 axis constitutes a posttranslational modulator of cell invasion whose aberration promotes cancer malignancy. Cancer Res; 77(9); 2328-38. ©2017 AACR.
Assuntos
Citoesqueleto de Actina/genética , Invasividade Neoplásica/genética , Neoplasias Pancreáticas/genética , Proteína 1 de Ligação a Repetições Teloméricas/genética , Citoesqueleto de Actina/patologia , Fatores de Despolimerização de Actina/genética , Proteína de Capeamento de Actina CapZ/genética , Adesão Celular/genética , Linhagem Celular Tumoral , Feminino , Adesões Focais/genética , Regulação Neoplásica da Expressão Gênica , Humanos , Quinases Lim/genética , Masculino , Invasividade Neoplásica/patologia , Neoplasias Pancreáticas/patologia , Fosforilação , Poli(ADP-Ribose) Polimerase-1/genética , Proteína 1 de Ligação a Repetições Teloméricas/biossíntese , Via de Sinalização Wnt , Quinases Associadas a rho/genéticaRESUMO
The expansion of a polyQ repeat within the ataxin-2 protein causes spinocerebellar ataxia type 2 (SCA2). However, neither the precise pathological mechanism nor the physiological functions of ataxin-2 are known. Ataxin-2 contains 47 (S/T)P sequences, which are targeted by proline-directed protein kinases such as the cyclin-dependent kinase 5 (Cdk5). We hypothesized that ataxin-2 is phosphorylated by Cdk5. In fact, phosphorylation of ataxin-2 by Cdk5-p25 was shown using two methods: in vitro(32)P labeling and electrophoretic mobility shift on Phos-tag SDS-PAGE. The fractionation of ataxin-2 into three portions, the N-terminal fragment (NF, amino acids 1-507), the middle fragment (MF, amino acids 508-905), and the C-terminal fragment (CF, amino acids 906-1313) showed that NF and MF were phosphorylated slightly and highly, respectively, by Cdk5-p25 when expressed in COS-7 cells. Cdk5-mediated phosphorylation induced the degradation of NF remarkably and MF moderately. Furthermore, toxic ataxin-2-41Q underwent proteasomal degradation after phosphorylation by Cdk5. These results suggest that Cdk5 controls the abundance of both normal and polyQ-expanded ataxin-2 protein in neurons, which implies that Cdk5 activity is a therapeutic approach for SCA2.