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Rescue of ALS Protein FUS Toxicity by TAF

Hayden, Elliott
http://rave.ohiolink.edu/etdc/view?acc_num=wright1559680404963737

Abstract Details

2019, Doctor of Philosophy (PhD), Wright State University, Biomedical Sciences PhD.
Amyotrophic Lateral Scleroses (ALS) is a neurodegenerative disease characterized by the degeneration of upper and lower motor neurons in the brain and spinal cord leading to progressive paralysis and ultimately death within 5 years of symptom onset. Only two drugs are approved by the Food and Drug Administration (FDA) for treating ALS that slow the disease progression by about 3 months. Mutations in the gene Fused in Sarcoma (FUS) cause inherited forms of ALS. FUS is a nuclear, multifunctional RNA-binding protein (RBP) involved in multiple RNA metabolic pathways. Mutations in FUS cause mislocalization of the protein from the nucleus to cytoplasm, where it forms inclusion that colocalize with stress granules. Over-expression of human FUS in the budding yeast, Saccharomyces cerevisiae, results in cellular toxicity, cytoplasmic aggregation and localization to stress granules, recapitulating phenotypes of mutant FUS in mammalian models and patients. In recent years, perturbations in RNA metabolism and RNA binding proteins has emerged as an underlying defect in ALS pathogenesis. FUS is one of the first RBPs linked to ALS, and modeling FUS toxicity in yeast will enhance our understanding of how RBPs contribute to neuronal toxicity in ALS. Using a yeast model of FUS toxicity, we designed and completed a genetic screen to identify human genes that suppress FUS induced toxicity. Enrichment analysis of the suppressor genes showed an over representation of genes with RNA binding function and ribonucleoprotein complex localization. A subset of the suppressors physically interact with FUS and colocalize with FUS aggregates. We focused on the FUS suppressor TATA-Box Binding Protein Associated Factor (TAF) for further study since TAF has been found in a complex with FUS and has a similar function and structure to FUS. We tested TAF against three other toxic neurodegenerative disease proteins; TAF suppressed toxicity of FUS and not the others. TAF did not reduce FUS protein expression or re-localize FUS from cytoplasmic foci to the nucleus. TAF and FUS interact with each other and colocalize with p-bodies and stress granules. The C-terminus of TAF is essential for its interaction with FUS. Deletion of TAF C-terminus eliminates its interaction with FUS and its ability to suppress toxicity, indicating that rescue is mediated through the interaction. TAF’s RNA recognition motif (RRM) is critical for rescue since mutation of phenylalanine residues within the RRM eliminates rescue. Wild type (WT) TAF but not TAF with a mutated RRM reduced the number of FUS aggregates. This indicates that TAF, through its RNA binding ability, is able to reduce FUS aggregates and therefore, stress granules. This study establishes that TAF, an interacting partner of FUS, can suppress FUS toxicity without decreasing FUS protein levels or altering cytoplasmic localization. As the suppressive effect of TAF is associated with reduction in the number of FUS stress granule, disassembling pathological stress granules may represent a therapeutic target.
Shulin Ju, Ph.D. (Advisor)
Paula Bubulya, Ph.D. (Committee Member)
Weiwen Long, Ph.D. (Committee Member)
Mark Rich, M.D., Ph.D. (Committee Member)
Quan Zhong, Ph.D. (Committee Member)
162 p.
Molecular Biology
yeast; neurodegeneration; ALS; protein aggregation; amyotrophic lateral sclerosis; stress granule; p-bodies

Recommended Citations

Citations

  • Hayden, E. (2019). Rescue of ALS Protein FUS Toxicity by TAF [Doctoral dissertation, Wright State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=wright1559680404963737

    APA Style (7th edition)

  • Hayden, Elliott. Rescue of ALS Protein FUS Toxicity by TAF. 2019. Wright State University, Doctoral dissertation. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=wright1559680404963737.

    MLA Style (8th edition)

  • Hayden, Elliott. "Rescue of ALS Protein FUS Toxicity by TAF." Doctoral dissertation, Wright State University, 2019. http://rave.ohiolink.edu/etdc/view?acc_num=wright1559680404963737

    Chicago Manual of Style (17th edition)

wright1559680404963737
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© 2019, all rights reserved.
This open access ETD is published by Wright State University and OhioLINK.