Distinct functional domains in nesprin-1α and nesprin-2β bind directly to emerin and both interactions are disrupted in X-linked Emery–Dreifuss muscular dystrophy

• Published in: Experimental cell research Vol. 313; no. 13; pp. 2845 - 2857
• Main Authors: Wheeler, Matthew A., Davies, John D., Zhang, Qiuping, Emerson, Lindsay J., Hunt, James, Shanahan, Catherine M., Ellis, Juliet A.
• Format: Journal Article
• Language: English
• Published: Elsevier Inc 01-08-2007
• Subjects: Emerin; Emery–Dreifuss muscular dystrophy; Nesprin-1 and -2; Emerin; Emery–Dreifuss muscular dystrophy; Nesprin-1 and -2
• ISSN: 0014-4827; 1090-2422
• DOI: 10.1016/j.yexcr.2007.03.025

Emerin and specific isoforms of nesprin-1 and -2 are nuclear membrane proteins which are binding partners in multi-protein complexes spanning the nuclear envelope. We report here the characterisation of the residues both in emerin and in nesprin-1α and -2β which are involved in their interaction and show that emerin requires nesprin-1 or -2 to retain it at the nuclear membrane. Using several protein–protein interaction methods, we show that residues 368 to 627 of nesprin-1α and residues 126 to 219 of nesprin-2β, which show high homology to one another, both mediate binding to emerin residues 140–176. This region has previously been implicated in binding to F-actin, β-catenin and lamin A/C suggesting that it is critical for emerin function. Confirmation that these protein domains interact in vivo was shown using GFP-dominant negative assays. Exogenous expression of either of these nesprin fragments in mouse myoblast C2C12 cells displaced endogenous emerin from the nuclear envelope and reduced the targeting of newly synthesised emerin. Furthermore, we are the first to report that emerin mutations which give rise to X-linked Emery–Dreifuss muscular dystrophy, disrupt binding to both nesprin-1α and -2β isoforms, further indicating a role of nesprins in the pathology of Emery–Dreifuss muscular dystrophy.