Abstract
Mutations in TARDBP, encoding TAR DNA-binding protein-43 (TDP-43), are associated with TDP-43 proteinopathies, including amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD). We compared wild-type TDP-43 and an ALS-associated mutant TDP-43 in vitro and in vivo. The A315T mutant enhances neurotoxicity and the formation of aberrant TDP-43 species, including protease-resistant fragments. The C terminus of TDP-43 shows sequence similarity to prion proteins. Synthetic peptides flanking residue 315 form amyloid fibrils in vitro and cause neuronal death in primary cultures. These data provide evidence for biochemical similarities between TDP-43 and prion proteins, raising the possibility that TDP-43 derivatives may cause spreading of the disease phenotype among neighboring neurons. Our work also suggests that decreasing the abundance of neurotoxic TDP-43 species, enhancing degradation or clearance of such TDP-43 derivatives and blocking the spread of the disease phenotype may have therapeutic potential for TDP-43 proteinopathies.
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Acknowledgements
We thank F. Baralle, E. Buratti, B. Cui, D. Kuo, N. Jayaram, Y. Li, M. Mishra, S. Perrett, M.-Y. Shen, M.-J. Zhang and T. Siddique for providing invaluable suggestions and reagents and for critical reading of the manuscript. We thank members of the Wu laboratory for stimulating discussions and suggestions. We thank L. Guo and L. Wang for technical assistance and A. Joselin for help in the early stage of the work. W.G. (grant 2009CB825402) and Y.C., H.Y. and Q.X. (grant 2010CB529603) are supported by the Ministry of Science and Technology (MOST) China 973 Project . J.Y.W. is supported by funds from Northwestern University and the Chinese Academy of Science (CAS). Y.Y. and C.W. are supported by CAS and MOST. We also thank the US National Institutes of Health (grant AG13854 to M.M. and E.H.B.) for support.
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J.Y.W., W.G., X.Z., K.F. and E.J.R. designed the study; W.G., Y.C., X.Z., A.K., P.R., X.C., E.J.R., M.Y., L.Z., J.L., M.X., Y.Y., C.W., D.Z., K.F., E.J.R. and J.Y.W. performed the experiments and analyzed the data; H.Y., L.Z., J.L., Y.S., K.F., Q.X. and J.Y.W. supervised the experiments and discussed and analyzed the data; E.H.B. and M.M. provided crucial tissue samples and revised the manuscript; W.G., E.J.R. and J.Y.W. wrote the paper.
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Supplementary Movie 1
Molecular dynamics simulation suggests that the 46mer TDP-43 peptides adopt multiple conformations including collapsed globular conformation at the N-terminal half and extended β-sheet conformation at the C-terminal region, in agreement with the Protscale analyses (Fig. 7). Molecular dynamics simulations were carried out on Wt and A315T mutant TDP-43 synthetic peptide (Q286-Q331) using TINKER, a software tool for molecular design3. The force field used in this molecular dynamics simulation was CHARMM-19, in combination with a statistical potential of mean force DOPE (discrete optimized protein energy)4. Non-bonded forces (electrostatic and van der Waals) were truncated at 5Å. After the TDP peptide was energy minimized and equilibrated for 100-ps with 1-fs time step, ten (peptide dimer) or thirty (monomer) independent 100-ps trajectories were produced using different random initial conditions. The canonical ensemble simulations were kept at a constant temperature of 300K. The amino-termini of both peptides are at the lower right corner, with the carboxyl termini at the upper left corner. The backbones of the wild type and A315T mutant peptides are shown in green and blue ribbons respectively with corresponding Ala315 or Thr315 residues marked in red. (MPG 430 kb)
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Guo, W., Chen, Y., Zhou, X. et al. An ALS-associated mutation affecting TDP-43 enhances protein aggregation, fibril formation and neurotoxicity. Nat Struct Mol Biol 18, 822–830 (2011). https://doi.org/10.1038/nsmb.2053
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DOI: https://doi.org/10.1038/nsmb.2053
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