RRC ID 52780
Author Kitamura A, Nakayama Y, Shibasaki A, Taki A, Yuno S, Takeda K, Yahara M, Tanabe N, Kinjo M.
Title Interaction of RNA with a C-terminal fragment of the amyotrophic lateral sclerosis-associated TDP43 reduces cytotoxicity.
Journal Sci Rep
Abstract A hallmark of amyotrophic lateral sclerosis (ALS), a devastating neurodegenerative disease, is formation of inclusion bodies (IBs) from misfolded proteins in neuronal cells. TAR RNA/DNA-binding protein 43 kDa (TDP43) is an ALS-causative protein forming IBs in ALS patients. The relation between localization of the IBs and neurotoxicity remains largely unknown. We characterized aggregation of fluorescently tagged TDP43 and its carboxyl-terminal fragments (CTFs) by analytical fluorescence imaging techniques. Quantitative time-lapse analysis in individual live cells showed that fluorescent-protein-tagged TDP43 was cleaved and a 35 kDa TDP43 CTF (TDP35) formed ubiquitin (Ub)-negative cytoplasmic IBs. Although TDP35 formed mildly toxic Ub-negative IBs in the cytoplasm, TDP25, another type of a TDP43 CTF, efficiently formed sufficiently toxic Ub-positive IBs. One- or two-color fluorescence correlation spectroscopy (FCS/FCCS) revealed that coaggregation of TDP25 with TDP43 was initiated by depletion of the RNA that binds to TDP25. Moreover, nuclear localization tagging TDP25 reduced the rate of neuronal cell death. These observations point to the need to elucidate the novel sequestration mechanism and details of the toxicity of the misfolded and aggregation-prone TDP43 CTFs (as well as the RNA binding and nuclear retention) in order to identify possible preventive interventions against ALS.
Volume 6
Pages 19230
Published 2016-1-13
DOI 10.1038/srep19230
PII srep19230
PMID 26757674
PMC PMC4725827
MeSH Amyotrophic Lateral Sclerosis / genetics Amyotrophic Lateral Sclerosis / metabolism* Animals Caspase 3 / metabolism Cell Death Cell Line DNA-Binding Proteins / chemistry DNA-Binding Proteins / metabolism* Humans Inclusion Bodies / metabolism Mice Peptide Fragments / metabolism Protein Aggregates Protein Aggregation, Pathological Protein Binding Protein Folding Protein Interaction Domains and Motifs* Protein Transport Proteolysis RNA / genetics RNA / metabolism*
IF 3.998
Times Cited 29
DNA material CMV:TDP43-GFP (RDB15742) CMV:GFP-TDP25 (RDB15743) CMV:GFP-NLS-TDP25 (RDB15744) CMV:GFP-NLSNP-TDP25 (RDB15745) GFP-NLS NP (RDB15746) mCherry-Ubiquitin (RDB15747).