Department of Genetics and Molecular Biology, Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional (CINVESTAV), 07360 Mexico City, Mexico.
Cell Biology Department, Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional (CINVESTAV), 07360 Mexico City, Mexico.
Int J Mol Sci. 2020 Aug 19;21(17):5944. doi: 10.3390/ijms21175944.
β-dystroglycan (β-DG) assembles with lamins A/C and B1 and emerin at the nuclear envelope (NE) to maintain proper nuclear architecture and function. To provide insight into the nuclear function of β-DG, we characterized the interaction between β-DG and emerin at the molecular level. Emerin is a major NE protein that regulates multiple nuclear processes and whose deficiency results in Emery-Dreifuss muscular dystrophy (EDMD). Using truncated variants of β-DG and emerin, via a series of in vitro and in vivo binding experiments and a tailored computational analysis, we determined that the β-DG-emerin interaction is mediated at least in part by their respective transmembrane domains (TM). Using surface plasmon resonance assays we showed that emerin binds to β-DG with high affinity (KD in the nanomolar range). Remarkably, the analysis of cells in which DG was knocked out demonstrated that loss of β-DG resulted in a decreased emerin stability and impairment of emerin-mediated processes. β-DG and emerin are reciprocally required for their optimal targeting within the NE, as shown by immunofluorescence, western blotting and immunoprecipitation assays using emerin variants with mutations in the TM domain and B-lymphocytes of a patient with EDMD. In summary, we demonstrated that β-DG plays a role as an emerin interacting partner modulating its stability and function.
β-肌营养不良聚糖(β-DG)与核纤层 A/C 和 B1 以及埃默林在核膜(NE)上组装,以维持适当的核结构和功能。为了深入了解β-DG 的核功能,我们在分子水平上对β-DG 和埃默林之间的相互作用进行了表征。埃默林是一种主要的核膜蛋白,调节多种核过程,其缺乏会导致埃默里-德雷福斯肌营养不良症(EDMD)。通过一系列体外和体内结合实验以及定制的计算分析,使用β-DG 和埃默林的截断变体,我们确定β-DG-埃默林相互作用至少部分由它们各自的跨膜结构域(TM)介导。使用表面等离子体共振分析,我们表明埃默林与β-DG 具有高亲和力(KD 在纳摩尔范围内)结合。值得注意的是,对 DG 敲除的细胞进行的分析表明,β-DG 的缺失导致埃默林稳定性降低,并损害了埃默林介导的过程。β-DG 和埃默林在 NE 内的最佳靶向中是相互需要的,如免疫荧光、Western blot 和使用 TM 结构域突变的埃默林变体以及 EDMD 患者的 B 淋巴细胞进行的免疫沉淀分析所示。总之,我们证明了β-DG 作为埃默林的相互作用伙伴发挥作用,调节其稳定性和功能。