Sorbonne Université, INSERM UMR_S974, Center for Research in Myology, 75013 Paris, France.
Laboratory of Structural Biology and Radiobiology, Institute for Integrative Biology of the Cell (CEA, CNRS, University Paris South), University Paris-Saclay, 91190 Gif-sur-Yvette, France.
Cells. 2019 Jun 10;8(6):570. doi: 10.3390/cells8060570.
Emerin is a nuclear envelope protein that contributes to genome organization and cell mechanics. Through its N-terminal LAP2-emerin-MAN1 (LEM)-domain, emerin interacts with the DNA-binding protein barrier-to-autointegration (BAF). Emerin also binds to members of the linker of the nucleoskeleton and cytoskeleton (LINC) complex. Mutations in the gene encoding emerin are responsible for the majority of cases of X-linked Emery-Dreifuss muscular dystrophy (X-EDMD). Most of these mutations lead to an absence of emerin. A few missense and short deletion mutations in the disordered region of emerin are also associated with X-EDMD. More recently, missense and short deletion mutations P22L, ∆K37 and T43I were discovered in emerin LEM-domain, associated with isolated atrial cardiac defects (ACD). Here we reveal which defects, at both the molecular and cellular levels, are elicited by these LEM-domain mutations. Whereas K37 mutation impaired the correct folding of the LEM-domain, P22L and T43I had no impact on the 3D structure of emerin. Surprisingly, all three mutants bound to BAF, albeit with a weaker affinity in the case of K37. In human myofibroblasts derived from a patient's fibroblasts, emerin ∆K37 was correctly localized at the inner nuclear membrane, but was present at a significantly lower level, indicating that this mutant is abnormally degraded. Moreover, SUN2 was reduced, and these cells were defective in producing actin stress fibers when grown on a stiff substrate and after cyclic stretches. Altogether, our data suggest that the main effect of mutation K37 is to perturb emerin function within the LINC complex in response to mechanical stress.
核膜蛋白emerin 有助于基因组组织和细胞力学。通过其 N 端 LAP2-emerin-MAN1(LEM)结构域,emerin 与 DNA 结合蛋白 barrier-to-autointegration(BAF)相互作用。emerin 还与核骨架和细胞骨架(LINC)复合物的成员结合。编码 emerin 的基因突变是导致大多数 X 连锁 Emery-Dreifuss 肌营养不良症(X-EDMD)的原因。这些突变大多数导致 emerin 缺失。在 emerin 的无序区域也存在少数错义突变和短缺失突变与 X-EDMD 相关。最近,在 emerin LEM 结构域中发现了错义突变和短缺失突变 P22L、∆K37 和 T43I,与孤立性心房心脏缺陷(ACD)相关。在这里,我们揭示了这些 LEM 结构域突变在分子和细胞水平上引起的缺陷。虽然 K37 突变会损害 LEM 结构域的正确折叠,但 P22L 和 T43I 对 emerin 的 3D 结构没有影响。令人惊讶的是,所有三种突变体都与 BAF 结合,尽管 K37 的亲和力较弱。在源自患者成纤维细胞的人肌成纤维细胞中,emerin ∆K37 正确定位于内核膜,但存在水平显著降低,表明该突变体异常降解。此外,SUN2 减少,这些细胞在刚性底物上生长和周期性拉伸后,无法产生肌动蛋白应力纤维。总之,我们的数据表明,突变 K37 的主要影响是在机械应力下扰乱 LINC 复合物内的 emerin 功能。