Isolation of human mitotic protein phosphatase complexes: identification of a complex between protein phosphatase 1 and the RNA helicase Ddx21

Veerle de Wever, David C. Lloyd, Isha Nasa, Mhairi Nimick, Laura Trinkle-Mulcahy, Robert Gourlay, Nick Morrice, Greg B. G. Moorhead

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    5 Citations (Scopus)

    Abstract

    Metazoan mitosis requires remodelling of sub-cellular structures to ensure proper division of cellular and genetic material. Faults often lead to genomic instability, cell cycle arrests and disease onset. These key structural changes are under tight spatial-temporal and post-translational control, with crucial roles for reversible protein phosphorylation. The phosphoprotein phosphatases PP1 and PP2A are paramount for the timely execution of mitotic entry and exit but their interaction partners and substrates are still largely unresolved. High throughput, mass-spectrometry based studies have limited sensitivity for the detection of low-abundance and transient complexes, a typical feature of many protein phosphatase complexes. Moreover, the limited timeframe during which mitosis takes place reduces the likelihood of identifying mitotic phosphatase complexes in asynchronous cells. Hence, numerous mitotic protein phosphatase complexes still await identification. Here we present a strategy to enrich and identify serine/threonine protein phosphatase complexes at the mitotic spindle. We thus identified a nucleolar RNA helicase, Ddx21/Gu, as a novel, direct PP1 interactor. Furthermore, our results place PP1 within the toposome, a Topoisomerase II alpha (TOPOIIa) containing complex with a key role in mitotic chromatin regulation and cell cycle progression, possibly via regulated protein phosphorylation. This study provides a strategy for the identification of further mitotic PP1 partners and the unravelling of PP1 functions during mitosis.
    Original languageEnglish
    Article numbere39510
    JournalPLoS ONE
    Volume7
    Issue number6
    DOIs
    Publication statusPublished - 2012

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    de Wever, V., Lloyd, D. C., Nasa, I., Nimick, M., Trinkle-Mulcahy, L., Gourlay, R., Morrice, N., & Moorhead, G. B. G. (2012). Isolation of human mitotic protein phosphatase complexes: identification of a complex between protein phosphatase 1 and the RNA helicase Ddx21. PLoS ONE, 7(6), [e39510]. https://doi.org/10.1371/journal.pone.0039510