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    • University of Missouri-Columbia
    • Graduate School - MU Theses and Dissertations (MU)
    • Theses and Dissertations (MU)
    • Dissertations (MU)
    • 2006 Dissertations (MU)
    • 2006 MU dissertations - Freely available online
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    Analysis of interactions between the germline RNA helicases (GLHs) and their regulators KGB-1 and CSN-5 in Caenorhabditis elegans

    Orsborn, April Marie, 1978-
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    [PDF] research.pdf (13.93Mb)
    Date
    2006
    Format
    Thesis
    Metadata
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    Abstract
    The Caenorhabditis elegans germline RNA helicases (GLHs) are constitutive components of P granules, non-membranous aggregates of protein and RNA that segregate with the nematode germline. The GLHs are critical for fertility. The novel MAP kinase, KGB-1, interacts with the GLH proteins, and the null kgb-1(um3) strain results in sterile worms at high temperatures; the germlines of these worms contain endomitotic replicating oocytes (EMO). We find that in kgb-1(um3) adults, while GLH-4 levels are similar to wild type, levels of GLH-1 are increased up to seven fold and the morphology of P granules is grossly affected. Binding of KGB-1 to GLH-1 requires a MAP kinase docking site. KGB-1 can phosphorylate GLH-1 using in vitro kinase assays, and GLH-1 is degraded by the proteasome in a KGB-1-dependent manner. In addition, KGB-1 physically associates with CSN-5 (COP9 signalosome subunit 5), another GLH binding partner. RNA interference (RNAi) of csn-5 results in sterile worms with under-proliferated germlines, mirroring the combined glh-1 and glh-4 RNAi phenotype. In contrast, elimination of csn-5 in the kgb-1(um3) background results in significantly more fertile worms than in non-injected kgb-1 worms. Based on these biochemical and genetic interactions, we propose KGB-1 and CSN-5 may oppositely regulate GLH-1, with KGB-1 degrading and CSN-5 protecting GLH-1. This cooperative system could maintain proper GLH-1 levels during normal germline development, as well as prevent excess GLH accumulation during the stressful conditions of high temperature and aging.
    URI
    https://doi.org/10.32469/10355/4499
    https://hdl.handle.net/10355/4499
    Degree
    Ph. D.
    Thesis Department
    Microbiology (Medicine) (MU)
    Rights
    OpenAccess.
    Collections
    • 2006 MU dissertations - Freely available online
    • Molecular Microbiology and Immunology electronic theses and dissertations (MU)

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