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Tryptophan 32-mediated SOD1 aggregation is attenuated by pyrimidine-like compounds in living cells

Journal Article


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Abstract


  • Over 160 mutations in superoxide dismutase 1 (SOD1) are associated with familial amyotrophic lateral sclerosis (fALS), where the main pathological feature is deposition of SOD1 into proteinaceous cytoplasmic inclusions. We previously showed that the tryptophan residue at position 32 (W32) mediates the prion-like propagation of SOD1 misfolding in cells, and that a W32S substitution blocks this phenomenon. Here, we used in vitro protein assays to demonstrate that a W32S substitution in SOD1-fALS mutants significantly diminishes their propensity to aggregate whilst paradoxically decreasing protein stability. We also show SOD1-W32S to be resistant to seeded aggregation, despite its high abundance of unfolded protein. A cell-based aggregation assay demonstrates that W32S substitution significantly mitigates inclusion formation. Furthermore, this assay reveals that W32 in SOD1 is necessary for the formation of a competent seed for aggregation under these experimental conditions. Following the observed importance of W32 for aggregation, we established that treatment of living cells with the W32-interacting 5-Fluorouridine (5-FUrd), and its FDA approved analogue 5-Fluorouracil (5-FU), substantially attenuate inclusion formation similarly to W32S substitution. Altogether, we highlight W32 as a significant contributor to SOD1 aggregation, and propose that 5-FUrd and 5-FU present promising lead drug candidates for the treatment of SOD1-associated ALS.

Publication Date


  • 2018

Citation


  • Pokrishevsky, E., McAlary, L., Farrawell, N. E., Zhao, B., Sher, M., Yerbury, J. J. & Cashman, N. R. (2018). Tryptophan 32-mediated SOD1 aggregation is attenuated by pyrimidine-like compounds in living cells. Scientific Reports, 8 (1), 15590-1-15590-12.

Scopus Eid


  • 2-s2.0-85055206467

Ro Full-text Url


  • https://ro.uow.edu.au/cgi/viewcontent.cgi?article=1363&context=smhpapers1

Ro Metadata Url


  • http://ro.uow.edu.au/smhpapers1/356

Start Page


  • 15590-1

End Page


  • 15590-12

Volume


  • 8

Issue


  • 1

Place Of Publication


  • United Kingdom

Abstract


  • Over 160 mutations in superoxide dismutase 1 (SOD1) are associated with familial amyotrophic lateral sclerosis (fALS), where the main pathological feature is deposition of SOD1 into proteinaceous cytoplasmic inclusions. We previously showed that the tryptophan residue at position 32 (W32) mediates the prion-like propagation of SOD1 misfolding in cells, and that a W32S substitution blocks this phenomenon. Here, we used in vitro protein assays to demonstrate that a W32S substitution in SOD1-fALS mutants significantly diminishes their propensity to aggregate whilst paradoxically decreasing protein stability. We also show SOD1-W32S to be resistant to seeded aggregation, despite its high abundance of unfolded protein. A cell-based aggregation assay demonstrates that W32S substitution significantly mitigates inclusion formation. Furthermore, this assay reveals that W32 in SOD1 is necessary for the formation of a competent seed for aggregation under these experimental conditions. Following the observed importance of W32 for aggregation, we established that treatment of living cells with the W32-interacting 5-Fluorouridine (5-FUrd), and its FDA approved analogue 5-Fluorouracil (5-FU), substantially attenuate inclusion formation similarly to W32S substitution. Altogether, we highlight W32 as a significant contributor to SOD1 aggregation, and propose that 5-FUrd and 5-FU present promising lead drug candidates for the treatment of SOD1-associated ALS.

Publication Date


  • 2018

Citation


  • Pokrishevsky, E., McAlary, L., Farrawell, N. E., Zhao, B., Sher, M., Yerbury, J. J. & Cashman, N. R. (2018). Tryptophan 32-mediated SOD1 aggregation is attenuated by pyrimidine-like compounds in living cells. Scientific Reports, 8 (1), 15590-1-15590-12.

Scopus Eid


  • 2-s2.0-85055206467

Ro Full-text Url


  • https://ro.uow.edu.au/cgi/viewcontent.cgi?article=1363&context=smhpapers1

Ro Metadata Url


  • http://ro.uow.edu.au/smhpapers1/356

Start Page


  • 15590-1

End Page


  • 15590-12

Volume


  • 8

Issue


  • 1

Place Of Publication


  • United Kingdom