The J domain of sacsin disrupts intermediate filament assembly

Abstract

Autosomal Recessive Spastic Ataxia of the Charlevoix Saguenay (ARSACS), is caused by loss of function mutations in the SACS gene, which encodes sacsin, a giant protein of 520 kDa. A key feature of the absence of sacsin in cells is the formation of abnormal bundles of intermediate filaments (IF) including neurofilaments (NF) in neurons and vimentin IF in fibroblasts, suggesting a role of sacsin in IF homeostasis. Sacsin contains a J domain (SacsJ) homologous to Hsp40, that can interact with Hsp70 chaperones. The SacsJ domain resolved NF bundles in cultured Sacs-/- neurons, however, its mechanism is still unclear. Here, we focused on the role of SacsJ in NF assembly. We report that the SacsJ domain directly interacts with NF proteins in vitro to disassemble NFL filaments, and to inhibit their initial assembly, in the absence of Hsp70. We generated a cell-penetrating peptide derived from this domain, SacsJ-myc-TAT, which was efficient in disassembling both vimentin IF and NF in cultured fibroblasts and Sacs+/+ motor neurons as well as NF bundles in cultured Sacs-/- motor neurons. Whereas a normal NF network was restored in Sacs-/- neurons treated with the SacsJ peptide, there was some loss of IF networks in Sacs+/+ fibroblasts or neurons. These results suggest that SacsJ is a key regulator of NF and IF networks in cells, with implications for its therapeutic use.

Genetics of Autosomal Recessive Spastic Ataxia of Charlevoix-Saguenay (ARSACS) and Role of Sacsin in Neurodegeneration

“…In Sacs–/– motor neurons, the NF bundles were resolved, and the NF network was restored [ 21 ]. These data suggest that the J-domain could impact the regulation of IF-NF assembly and disassembly directly [ 3 , 20 , 31 ].…”

“…Cellular and animal model studies were investigated to define the pathways related to sacsin dysfunctions. Girard et al (2012) found that fibroblasts from ARSACS patients presented abnormal mitochondrial functions [ 31 ]. Later, Pilliod et al suggested that monitoring mitochondrial abnormalities could be a possible diagnostic biomarker in ARSACS patients [ 35 , 36 ].…”