Identification and characterization of AtI-2, an Arabidopsis homologue of an ancient protein phosphatase 1 (PP1) regulatory subunit

George W. Templeton, Mhairi Nimick, Nicholas Morrice, David Campbell, Marilyn Goudreault, Anne-Claude Gingras, Atsushi Takemiya, Ken-ichiro Shimazaki, Greg B. G. Moorhead

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    Abstract

    PP1 (protein phosphatase 1) is among the most conserved enzymes known, with one or more isoforms present in all sequenced eukaryotic genomes. PP1 dephosphorylates specific serine/threonine phosphoproteins as defined by associated regulatory or targeting subunits. In the present study we performed a PP1-binding screen to find putative PP1 interactors in Arabidopsis thaliana and uncovered a homologue of the ancient PP1 interactor, I-2 (inhibitor-2). Bioinformatic analysis revealed remarkable conservation of three regions of plant I-2 that play key roles in binding to PP1 and regulating its function. The sequence-related properties of plant I-2 were compared across eukaryotes, indicating a lack of I-2 in some species and the emergence points from key motifs during the evolution of this ancient regulator. Biochemical characterization of AtI-2 (Arabidopsis I-2) revealed its ability to inhibit all plant PP1 isoforms and inhibitory dependence requiring the primary interaction motif known as RVXF. Arabidopsis I-2 was shown to be a phosphoprotein in vivo that was enriched in the nucleus. TAP (tandem affinity purification)-tag experiments with plant I-2 showed in vivo association with several Arabidopsis PP1 isoforms and identified other potential I-2 binding proteins.

    Original languageEnglish
    Pages (from-to)73-83
    Number of pages11
    JournalBiochemical Journal
    Volume435
    DOIs
    Publication statusPublished - 1 Apr 2011

    Cite this

    Templeton, G. W., Nimick, M., Morrice, N., Campbell, D., Goudreault, M., Gingras, A-C., Takemiya, A., Shimazaki, K., & Moorhead, G. B. G. (2011). Identification and characterization of AtI-2, an Arabidopsis homologue of an ancient protein phosphatase 1 (PP1) regulatory subunit. Biochemical Journal, 435, 73-83. https://doi.org/10.1042/BJ20101035