Trends in Understanding the Pathological Roles of TDP-43 and FUS Proteins

Antisense oligonucleotide silencing of FUS expression as a therapeutic approach in amyotrophic lateral sclerosis

“…Like TDP-43, MATR3 and hnRNP A1, to which it is structurally and functionally related, FUS is one of several RNA-binding proteins (RBPs) that have been implicated in ALS. It is a ubiquitously expressed, predominantly nuclear protein that functions in DNA repair and several aspects of RNA metabolism, including transcription, pre-mRNA splicing, mRNA transport, stability and translation as well as the processing of microRNAs and other non-coding RNAs 9 . To date, more than 50 different FUS mutations have been identified in patients with ALS 10 , which together account for approximately 4% of familial cases and fewer than 2% of patients with sporadic ALS 10 , 11 .…”

Unraveling the toxic effects mediated by the neurodegenerative disease–associated S375G mutation of TDP-43 and its S375E phosphomimetic variant

“…Production of the TDP-43 C-terminal regions with the WT sequence and carrying the S375G and S375E mutants has been carried out as previously described (100). Briefly, CTD constructs with an N-terminal hexa histidine tag were expressed in Escherichia coli BL21 star cells in M9 media supplemented with 15 NH 4 Cl and 13 C-glucose and purified using nickelnitrilotriacetic acid agarose beads that were washed with denaturing buffer (20 mM Tris-Cl, 500 mM NaCl, 10 mM imidazole, 1 mM DTT, 8 M urea, pH 8.0), with a final elution using an imidazole gradient (26). The purified polypeptides were applied into a PD-10 desalting column pre-equilibrated with 1 mM DAc to obtain a final concentration of 10 μM and at pH = 4.0.…”

“…In TDP-43 proteinopathies, the protein is often aggregating in the cellular cytoplasm (although nuclear aggregates can also occur) and undergoes different post-translational modifications (PTMs), such as ubiquitination, phosphorylation, acetylation, sumoylation, and cleavage, to yield C-terminal fragments ( 13 ). Several studies performed on PTMs have highlighted their ability to regulate the interaction profile of TDP-43 with RNA and protein substrates and act as an important quality control checkpoint ( 14 ).…”

Parkin beyond Parkinson’s Disease—A Functional Meaning of Parkin Downregulation in TDP-43 Proteinopathies

“…TAR DNA-Binding Protein-43 (TDP-43) was first identified in 2001 as a protein able to bind the HIV-1 TAR binding sequence [ 61 ], and in 2006, as the main component of aggregates found in the brains of patients with ALS and FTLD [ 11 , 19 ] ( Figure 1 ). Recently, many reviews have been focused on elucidating the role of this protein in disease and normal development, and, for this reason, the reader is referred to these works for a more detailed description [ 10 , 62 , 63 , 64 , 65 ]. Briefly, TDP-43 belongs to the class of heterogeneous ribonucleoproteins (hnRNPs) that have been long referred to as the “RNA histones”.…”