Multiple Isoforms of Nesprin1 Are Integral Components of Ciliary Rootlets

SYNE1 (synaptic nuclear envelope 1) encodes multiple isoforms of Nesprin1 (nuclear envelope spectrin 1) that associate with the nuclear envelope (NE) through a C-terminal KASH (Klarsicht/Anc1/Syne homology) domain (Figure 1A) [1–4]. This domain interacts directly with the SUN (Sad1/Unc84) domain of Sun proteins [5–7], a family of transmembrane proteins of the inner nuclear membrane (INM) [8, 9], to form the so-called LINC complexes (linkers of the nucleoskeleton and cytoskeleton) that span the entire NE and mediate nuclear positioning [10–12]. In a stark departure from this classical depiction of Nesprin1 in the context of the NE, we report here that rootletin recruits Nesprin1α at the ciliary rootlets of photoreceptors and identify asymmetric NE aggregates of Nesprin1α and Sun2 that dock filaments of rootletin at the nuclear surface. In NIH 3T3 cells, we show that recombinant rootletin filaments also dock to the NE through the specific recruitment of an ∼600-kDa endogenous isoform of Nesprin1 (Nes1600kDa) and of Sun2. In agreement with the association of Nesprin1α with photoreceptor ciliary rootlets and the functional interaction between rootletin and Nesprin1 in fibroblasts, we demonstrate that multiple isoforms of Nesprin1 are integral components of ciliary rootlets of multiciliated ependymal and tracheal cells. Together, these data provide a novel functional paradigm for Nesprin1 at ciliary rootlets and suggest that the wide spectrum of human pathologies linked to truncating mutations of SYNE1 [13–15] may originate in part from ciliary defects. •Nesprin1 localizes to ciliary rootlets of photoreceptors and multiciliated cells•Nesprin1 docks rootletin filaments at the nuclear surface•Rootletin filaments induce the aggregation of SUN2/Nesprin1 LINC complexes•Loss of function of ciliary Nesprin1 may underlie multisystemic human pathologies Potter et al. report a novel interaction between Nesprin1 and rootletin that underlies the localization of multiple isoforms of Nesprin1 to ciliary rootlets and the docking of rootletin filaments at the nuclear surface by Sun2/Nesprin1 LINC complexes. This new paradigm may explain how mutations of SYNE1 underlie multisystemic human pathologies.