Specific interaction with the nuclear transporter importin α2 can modulate paraspeckle protein 1 delivery to nuclear paraspeckles

Andrew T. Major, Cathryn A. Hogarth, Yoichi Miyamoto, Mai A. Sarraj, Catherine L. Smith, Peter Koopman, Yasuyuki Kurihara, David A. Jans, Kate L. Loveland

    • 1 Citations

    Abstract

    Importin (IMP) superfamily members mediate regulated nucleocytoplasmic transport, which is central to key cellular processes. Although individual IMPa proteins exhibit dynamic synthesis and subcellular localization during cellular differentiation, including during spermatogenesis, little is known of how this affects cell fate. To investigate how IMPas control cellular development, we conducted a yeast two-hybrid screen for IMPα2 cargoes in embryonic day 12.5 mouse testis, a site of peak IMPα2 expression coincident with germ-line masculization. We identified paraspeckle protein 1(PSPC1), the original defining component of nuclear paraspeckles, as an IMPα2-binding partner. PSPC1-IMPα2 binding in testis was confirmed in immunoprecipitations and pull downs, and an enzyme-linked immunosorbent assay-based assay demonstrated direct, high-affinity PSPC1 binding to either IMPα2/IMPβ1 or IMPα6/IMPβ1. Coexpression of full-length PSPC1 and IMPα2 in HeLa cells yielded increased PSPC1 localization in nuclear paraspeckles. High-throughput image analysis of >3500 cells indicated IMPα2 levels can directly determine PSPC1-positive nuclear speckle numbers and size; a transport-deficient IMPα2 isoform or small interfering RNA knockdown of IMPα2 each reduced endogenous PSPC1 accumulation in speckles. This first validation of an IMPα2 nuclear import cargo in fetal testis provides novel evidence that PSPC1 delivery to paraspeckles, and consequently paraspeckle function, may be controlled by modulated synthesis of specific IMPs.

    Original languageEnglish
    Pages (from-to)1543-1558
    Number of pages16
    JournalMolecular Biology of the Cell
    Volume26
    Issue number8
    DOIs
    StatePublished - 2015 Apr 15

    Fingerprint

    Karyopherins
    Proteins
    Testis
    Cell Nucleus Active Transport
    Inosine Monophosphate
    Spermatogenesis
    HeLa Cells
    Immunoprecipitation
    Protein Binding
    Germ Cells
    Small Interfering RNA
    Protein Isoforms
    Yeasts
    Enzyme-Linked Immunosorbent Assay

    ASJC Scopus subject areas

    • Molecular Biology
    • Cell Biology

    Cite this

    Major, A. T., Hogarth, C. A., Miyamoto, Y., Sarraj, M. A., Smith, C. L., Koopman, P., ... Loveland, K. L. (2015). Specific interaction with the nuclear transporter importin α2 can modulate paraspeckle protein 1 delivery to nuclear paraspeckles. Molecular Biology of the Cell, 26(8), 1543-1558. DOI: 10.1091/mbc.E14-01-0678

    Specific interaction with the nuclear transporter importin α2 can modulate paraspeckle protein 1 delivery to nuclear paraspeckles. / Major, Andrew T.; Hogarth, Cathryn A.; Miyamoto, Yoichi; Sarraj, Mai A.; Smith, Catherine L.; Koopman, Peter; Kurihara, Yasuyuki; Jans, David A.; Loveland, Kate L.

    In: Molecular Biology of the Cell, Vol. 26, No. 8, 15.04.2015, p. 1543-1558.

    Research output: Contribution to journalArticle

    Major, AT, Hogarth, CA, Miyamoto, Y, Sarraj, MA, Smith, CL, Koopman, P, Kurihara, Y, Jans, DA & Loveland, KL 2015, 'Specific interaction with the nuclear transporter importin α2 can modulate paraspeckle protein 1 delivery to nuclear paraspeckles' Molecular Biology of the Cell, vol 26, no. 8, pp. 1543-1558. DOI: 10.1091/mbc.E14-01-0678
    Major AT, Hogarth CA, Miyamoto Y, Sarraj MA, Smith CL, Koopman P et al. Specific interaction with the nuclear transporter importin α2 can modulate paraspeckle protein 1 delivery to nuclear paraspeckles. Molecular Biology of the Cell. 2015 Apr 15;26(8):1543-1558. Available from, DOI: 10.1091/mbc.E14-01-0678

    Major, Andrew T.; Hogarth, Cathryn A.; Miyamoto, Yoichi; Sarraj, Mai A.; Smith, Catherine L.; Koopman, Peter; Kurihara, Yasuyuki; Jans, David A.; Loveland, Kate L. / Specific interaction with the nuclear transporter importin α2 can modulate paraspeckle protein 1 delivery to nuclear paraspeckles.

    In: Molecular Biology of the Cell, Vol. 26, No. 8, 15.04.2015, p. 1543-1558.

    Research output: Contribution to journalArticle

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