Zhang Q, Skepper J N, Yang F, Davies J D, Hegyi L, Roberts R G, Weissberg P L, Ellis J A, Shanahan C M
Department of Medicine, Division of Cardiovascular Medicine, University of Cambridge, Box 110, Addenbrooke's Hospital, Hills Road, Cambridge, CB2 2QQ, UK.
J Cell Sci. 2001 Dec;114(Pt 24):4485-98. doi: 10.1242/jcs.114.24.4485.
In search of vascular smooth muscle cell differentiation markers, we identified two genes encoding members of a new family of type II integral membrane proteins. Both are ubiquitously expressed, and tissue-specific alternative mRNA initiation and splicing generate at least two major isoforms of each protein, with the smaller isoforms being truncated at the N-terminus. We have named these proteins nesprin-1 and -2 for nuclear envelope spectrin repeat, as they are characterized by the presence of multiple, clustered spectrin repeats, bipartite nuclear localization sequences and a conserved C-terminal, single transmembrane domain. Transient transfection of EGFP-fusion expression constructs demonstrated their localization to the nuclear membrane with a novel C-terminal, TM-domain-containing sequence essential for perinuclear localization. Using antibodies to nesprin-1, we documented its colocalization with LAP1, emerin and lamins at the nuclear envelope, and immunogold labeling confirmed its presence at the nuclear envelope and in the nucleus where it colocalized with heterochromatin. Nesprin-1 is developmentally regulated in both smooth and skeletal muscle and is re-localized from the nuclear envelope to the nucleus and cytoplasm during C2C12 myoblast differentiation. These data and structural analogies with other proteins suggest that nesprins may function as 'dystrophins of the nucleus' to maintain nuclear organization and structural integrity.
为寻找血管平滑肌细胞分化标志物,我们鉴定出两个基因,它们编码一个新的II型整合膜蛋白家族的成员。二者均在全身表达,且组织特异性的可变mRNA起始和剪接产生了每种蛋白质的至少两种主要异构体,较小的异构体在N端被截短。我们将这些蛋白质命名为nesprin-1和nesprin-2,即核膜血影蛋白重复序列蛋白,因为它们的特征是存在多个成簇的血影蛋白重复序列、双分型核定位序列以及保守的C端单跨膜结构域。EGFP融合表达构建体的瞬时转染表明,它们定位于核膜,其C端含TM结构域的新序列对核周定位至关重要。使用针对nesprin-1的抗体,我们证明了它与LAP1、emerin和核纤层蛋白在核膜处共定位,免疫金标记证实了它在核膜以及与异染色质共定位的细胞核中的存在。Nesprin-1在平滑肌和骨骼肌中均受到发育调控,并且在C2C12成肌细胞分化过程中从核膜重新定位于细胞核和细胞质。这些数据以及与其他蛋白质的结构相似性表明,nesprins可能作为“细胞核的肌营养不良蛋白”发挥作用,以维持核组织和结构完整性。