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Deconvoluting Stress-Responsive Proteostasis Signaling Pathways for Pharmacologic Activation Using Targeted RNA Sequencing

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    Deconvoluting Stress-Responsive Proteostasis Signaling Pathways for Pharmacologic Activation Using Targeted RNA Sequencing
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    • Julia M. D. Grandjean
      Julia M. D. Grandjean
      Department of Molecular Medicine, The Scripps Research Institute, La Jolla, California 92037, United States
      More by Julia M. D. Grandjean
    • Lars Plate
      Lars Plate
      Department of Molecular Medicine  and  Department of Chemistry, The Scripps Research Institute, La Jolla, California 92037, United States
      More by Lars Plate
      Orcidhttp://orcid.org/0000-0003-4363-6116
    • Richard I. Morimoto
      Richard I. Morimoto
      Department of Molecular Biosciences, Rice Institute for Biomedical Research, Northwestern University, Evanston, Illinois 60208, United States
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    • Michael J. Bollong
      Michael J. Bollong
      Department of Chemistry, The Scripps Research Institute, La Jolla, California 92037, United States
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      Orcidhttp://orcid.org/0000-0001-9439-1476
    • Evan T. Powers
      Evan T. Powers
      Department of Chemistry, The Scripps Research Institute, La Jolla, California 92037, United States
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    • R. Luke Wiseman*
      R. Luke Wiseman
      Department of Molecular Medicine, The Scripps Research Institute, La Jolla, California 92037, United States
      *Department of Molecular Medicine, The Scripps Research Institute, La Jolla, CA 92037. E-mail: [email protected]. Phone: (858) 784-8820.
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      Orcidhttp://orcid.org/0000-0001-9287-6840
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    ACS Chemical Biology

    Cite this: ACS Chem. Biol. 2019, 14, 4, 784–795
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    https://doi.org/10.1021/acschembio.9b00134
    Published March 1, 2019
    Copyright © 2019 American Chemical Society
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    Abstract

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    Cellular proteostasis is maintained by stress-responsive signaling pathways such as the heat shock response (HSR), the oxidative stress response (OSR), and the unfolded protein response (UPR). Activation of these pathways results in the transcriptional upregulation of select subsets of stress-responsive genes that restore proteostasis and adapt cellular physiology to promote recovery following various types of acute insult. The capacity for these pathways to regulate cellular proteostasis makes them attractive therapeutic targets for correcting proteostasis defects associated with diverse diseases. High-throughput screening (HTS) using cell-based reporter assays is highly effective for identifying putative activators of stress-responsive signaling pathways. However, the development of these compounds is hampered by the lack of medium-throughput assays to define compound potency and selectivity for a given pathway. Here, we describe a targeted RNA sequencing (RNAseq) assay that allows cost-effective, medium-throughput screening of stress-responsive signaling pathway activation. We demonstrate that this assay allows deconvolution of stress-responsive signaling activated by chemical genetic or pharmacologic agents. Furthermore, we use this assay to define the selectivity of putative OSR and HSR activating compounds previously identified by HTS. Our results demonstrate the potential for integrating this adaptable targeted RNAseq assay into screening programs focused on developing pharmacologic activators of stress-responsive signaling pathways.

    Copyright © 2019 American Chemical Society

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    Supporting Information

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    The Supporting Information is available free of charge on the ACS Publications website at DOI: 10.1021/acschembio.9b00134.

    • Supplemental experimental procedures, figure legends for Figures S1–S7, Figures S1–S7, supplemental table legends for Tables S1–S5, and Table S3 (PDF)

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    ACS Chemical Biology

    Cite this: ACS Chem. Biol. 2019, 14, 4, 784–795
    Click to copy citationCitation copied!
    https://doi.org/10.1021/acschembio.9b00134
    Published March 1, 2019
    Copyright © 2019 American Chemical Society
    Request reuse permissions

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