|A yeast model of FUS/TLS-dependent cytotoxicity.
|Year of Publication
|Ju S, Tardiff DF, Han H, Divya K, Zhong Q, Maquat LE, Bosco DA, Hayward LJ, Brown RH, Lindquist S, Ringe D, Petsko GA
|Amyotrophic Lateral Sclerosis, Cell Nucleus, Cytoplasm, DNA Helicases, Gene Expression Regulation, Mutation, Neurons, Recombinant Fusion Proteins, RNA, Messenger, RNA-Binding Protein FUS, Saccharomyces cerevisiae, Saccharomyces cerevisiae Proteins, Trans-Activators
FUS/TLS is a nucleic acid binding protein that, when mutated, can cause a subset of familial amyotrophic lateral sclerosis (fALS). Although FUS/TLS is normally located predominantly in the nucleus, the pathogenic mutant forms of FUS/TLS traffic to, and form inclusions in, the cytoplasm of affected spinal motor neurons or glia. Here we report a yeast model of human FUS/TLS expression that recapitulates multiple salient features of the pathology of the disease-causing mutant proteins, including nuclear to cytoplasmic translocation, inclusion formation, and cytotoxicity. Protein domain analysis indicates that the carboxyl-terminus of FUS/TLS, where most of the ALS-associated mutations are clustered, is required but not sufficient for the toxicity of the protein. A genome-wide genetic screen using a yeast over-expression library identified five yeast DNA/RNA binding proteins, encoded by the yeast genes ECM32, NAM8, SBP1, SKO1, and VHR1, that rescue the toxicity of human FUS/TLS without changing its expression level, cytoplasmic translocation, or inclusion formation. Furthermore, hUPF1, a human homologue of ECM32, also rescues the toxicity of FUS/TLS in this model, validating the yeast model and implicating a possible insufficiency in RNA processing or the RNA quality control machinery in the mechanism of FUS/TLS mediated toxicity. Examination of the effect of FUS/TLS expression on the decay of selected mRNAs in yeast indicates that the nonsense-mediated decay pathway is probably not the major determinant of either toxicity or suppression.
|PubMed Central ID
|1RC1NS06839 / NS / NINDS NIH HHS / United States
1RC2NS070342-01 / NS / NINDS NIH HHS / United States
NS614192 / NS / NINDS NIH HHS / United States
R01NS050557-05 / NS / NINDS NIH HHS / United States
U01NS05225-03 / NS / NINDS NIH HHS / United States
/ / Howard Hughes Medical Institute / United States