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    Identification of a small molecule yeast TORC1 inhibitor with a flow cytometry-based multiplex screen

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    Chen_ACSChemBiol_2012.pdf (2.538Mb)
    Issue Date
    2012-04-20
    Author
    Chen, Jun
    Young, Susan M.
    Allen, Chris
    Seeber, Andrew
    Péli-Gulli, Marie-Pierre
    Panchaud, Nicolas
    Waller, Anna
    Ursu, Oleg
    Yao, Tuanli
    Golden, Jennifer E.
    Strouse, J. Jacob
    Carter, Mark B.
    Kang, Huining
    Bologa, Cristian G.
    Foutz, Terry D.
    Edwards, Bruce S.
    Peterson, Blake R.
    Aubé, Jeffrey
    Werner-Washburne, Margaret
    Loewith, Robbie J.
    De Virgilio, Claudio
    Sklar, Larry A.
    Publisher
    ACS Chem Biol.
    Type
    Article
    Article Version
    Scholarly/refereed, author accepted manuscript
    Rights
    © 2012 American Chemical Society
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    Abstract
    TOR (target of rapamycin) is a serine/threonine kinase, evolutionarily conserved from yeast to human, which functions as a fundamental controller of cell growth. The moderate clinical benefit of rapamycin in mTOR-based therapy of many cancers favors the development of new TOR inhibitors. Here we report a high throughput flow cytometry multiplexed screen using five GFPtagged yeast clones that represent the readouts of four branches of the TORC1 signaling pathway in budding yeast. Each GFP-tagged clone was differentially color-coded and the GFP signal of each clone was measured simultaneously by flow cytometry, which allows rapid prioritization of compounds that likely act through direct modulation of TORC1 or proximal signaling components. A total of 255 compounds were confirmed in dose-response analysis to alter GFP expression in one or more clones. To validate the concept of the high throughput screen, we have characterized CID 3528206, a small molecule most likely to act on TORC1 as it alters GFP expression in all five GFP clones in an analogous manner to rapamycin. We have shown that CID 3528206 inhibited yeast cell growth, and that CID 3528206 inhibited TORC1 activity both in vitro and in vivo with EC50s of 150 nM and 3.9 μM, respectively. The results of microarray analysis and yeast GFP collection screen further support the notion that CID 3528206 and rapamycin modulate similar cellular pathways. Together, these results indicate that the HTS has identified a potentially useful small molecule for further development of TOR inhibitors.
    URI
    http://hdl.handle.net/1808/23523
    DOI
    https://doi.org/10.1021/cb200452r
    Collections
    • Medicinal Chemistry Scholarly Works [242]
    Citation
    Chen, J., Young, S. M., Allen, C., Seeber, A., Péli-Gulli, M.-P., Panchaud, N., … Sklar, L. A. (2012). Identification of a small molecule yeast TORC1 inhibitor with a flow cytometry-based multiplex screen. ACS Chemical Biology, 7(4), 715–722. http://doi.org/10.1021/cb200452r

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    Contact KU ScholarWorks
    785-864-8983
    KU Libraries
    1425 Jayhawk Blvd
    Lawrence, KS 66045
    785-864-8983

    KU Libraries
    1425 Jayhawk Blvd
    Lawrence, KS 66045
    Image Credits
     

     

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