Departamento de Genética y Biología Molecular, Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional, Ciudad de México 07360, Mexico.
Departamento de Bioquímica, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, Ciudad de México 11340, Mexico.
Int J Mol Sci. 2020 Jul 14;21(14):4961. doi: 10.3390/ijms21144961.
Nuclear β-dystroglycan (β-DG) is involved in the maintenance of nuclear architecture and function. Nonetheless, its relevance in defined nuclear processes remains to be determined. In this study we generated a C2C12 cell-based DG-null model using CRISPR-Cas9 technology to provide insights into the role of β-DG on nuclear processes. Since DG-null cells exhibited decreased levels of lamin B1, we aimed to elucidate the contribution of DG to senescence, owing to the central role of lamin B1 in this pathway. Remarkably, the lack of DG enables C2C12 cells to acquire senescent features, including cell-cycle arrest, increased senescence-associated-β-galactosidase activity, heterochromatin loss, aberrant nuclear morphology and nucleolar disruption. We demonstrated that genomic instability is one driving cause of the senescent phenotype in DG-null cells via the activation of a DNA-damage response associated with mitotic failure, as shown by the presence of multipolar mitotic spindles, which in turn induced the formation of micronuclei and γH2AX foci (DNA-damage marker), telomere shortening and p53/p21 upregulation. Altogether, these events might ultimately lead to premature senescence, impeding the replication of the damaged genome. In summary, we present evidence supporting a role for DG in protecting against senescence, through the maintenance of proper lamin B1 expression/localization and proper mitotic spindle organization.
核β-肌营养不良聚糖(β-DG)参与核架构和功能的维持。尽管如此,其在特定核过程中的相关性仍有待确定。在这项研究中,我们使用 CRISPR-Cas9 技术生成了一个基于 C2C12 细胞的 DG 缺失模型,以深入了解β-DG 在核过程中的作用。由于 DG 缺失细胞表现出 lamin B1 水平降低,我们旨在阐明 DG 对衰老的贡献,因为 lamin B1 在该途径中起着核心作用。值得注意的是,缺乏 DG 使 C2C12 细胞能够获得衰老特征,包括细胞周期停滞、衰老相关-β-半乳糖苷酶活性增加、异染色质丢失、核形态异常和核仁破坏。我们证明基因组不稳定性是 DG 缺失细胞衰老表型的一个驱动因素,这是通过与有丝分裂失败相关的 DNA 损伤反应的激活来实现的,这表现为多极有丝分裂纺锤体的存在,进而诱导微核和γH2AX 焦点(DNA 损伤标志物)的形成、端粒缩短和 p53/p21 的上调。总而言之,这些事件最终可能导致过早衰老,从而阻碍受损基因组的复制。总之,我们提供的证据支持 DG 通过维持适当的 lamin B1 表达/定位和适当的有丝分裂纺锤体组织来防止衰老。