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    A high-throughput screen for chemical inhibitors of exocytic transport in yeast

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    Zhang_Wiley_2010.pdf (982.7Kb)
    Issue Date
    2010-06-14
    Author
    Zhang, Lisha
    Nebane, N. Miranda
    Wennerberg, Krister
    Li, Yujie
    Neubauer, Valerie
    Hobarth, Judith V.
    McKellip, Sara N.
    Rasmussen, Lynn
    Shindo, Nice
    Sosa, Melinda
    Maddry, Joseph A.
    Ananthan, Subramaniam
    Piazza, Gary A.
    White, E. Lucile
    Harsay, Edina
    Publisher
    Wiley
    Type
    Article
    Article Version
    Scholarly/refereed, author accepted manuscript
    Rights
    © 2010 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
    Metadata
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    Abstract
    Most of the components of the membrane and protein traffic machinery were discovered by perturbing their functions, either with bioactive compounds or by mutations. However, the mechanisms responsible for exocytic transport vesicle formation at the Golgi and endosomes are still largely unknown. Both the exocytic traffic routes and the signaling pathways that regulate these routes are highly complex and robust, so that defects can be overcome by alternate pathways or mechanisms. A classical yeast genetic screen designed to account for the robustness of the exocytic pathway identified a novel conserved gene, AVL9, that functions in late exocytic transport. We now describe a chemical-genetic version of the mutant screen, in which we performed a high-throughput phenotypic screen of a large compound library and identified novel small molecule secretory inhibitors. In order to maximize the number and diversity of our hits, the screen was performed in a pdr5Δ snq2Δ mutant background, which lacks two transporters responsible for pleiotropic drug resistance. However, we found that deletion of both transporters reduced the fitness of our screen strain, whereas the pdr5Δ mutation had relatively small effect on growth and was also the more important transporter mutation for conferring sensitivity to our hits. In this and similar chemical-genetic yeast screens, using just a single pump mutation may be sufficient for increasing hit diversity while minimizing the physiological effects of transporter mutations.
    Description
    This is the peer reviewed version of the following article: Zhang, L., Nebane, N. M., Wennerberg, K., Li, Y., Neubauer, V., Hobrath, J. V., … Harsay, E. (2010). A high-throughput screen for chemical inhibitors of exocytic transport in yeast. Chembiochem : A European Journal of Chemical Biology, 11(9), 1291–1301. http://doi.org/10.1002/cbic.200900681, which has been published in final form at doi.org/10.1002/cbic.200900681. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving.
    URI
    http://hdl.handle.net/1808/24012
    DOI
    https://doi.org/10.1002/cbic.200900681
    Collections
    • Molecular Biosciences Scholarly Works [537]
    Citation
    Zhang, L., Nebane, N. M., Wennerberg, K., Li, Y., Neubauer, V., Hobrath, J. V., … Harsay, E. (2010). A high-throughput screen for chemical inhibitors of exocytic transport in yeast. Chembiochem : A European Journal of Chemical Biology, 11(9), 1291–1301. http://doi.org/10.1002/cbic.200900681

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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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