Spatio-temporal composition of the mitotic Chromosomal Passenger Complex detected using in situ proximity ligation assay: Technical Note

Mariaana Vuoriluoto, Leena Laine, Petri Saviranta, Jeroen Pouwels, Marko Kallio

    Research output: Contribution to journalArticlepeer-review

    4 Citations (Scopus)

    Abstract

    Cell division is orchestrated by a complex protein network that aims to maintenance of genomic stability. Visualisation of mitotic protein–protein associations in space and time has been limited due to the lack of proper biochemical and easy‐to‐use imaging tools. Here we report adaptation of the in situ proximity ligation assay (is‐PLA) to study mitotic protein interactions with spatio‐temporal resolution. We examined the composition of the Chromosomal Passenger Complex (CPC) at various mitotic phases and after chemical treatments using is‐PLA with antibodies against the core CPC subunits Aurora B, INCENP, Survivin and Borealin. Our results support the notion that the core CPC functions as a single structural unit at centromeres in early mitosis and at central spindle after the onset of anaphase. Treatment of cells with the Aurora B inhibitor ZM447439 diminished the is‐PLA signals at centromeres suggesting that Aurora B activity contributes to structural maintenance and/or proper subcellular localization of the core CPC. Is‐PLA‐based analysis of interaction between INCENP and Polo‐like kinase 1 (Plk1) proposes that the kinase co‐travels with CPC during late mitosis. The data illustrates both the strengths and limitations of the is‐PLA in the analysis of mitotic macromolecule associations at sub‐organelle level.
    Original languageEnglish
    Pages (from-to)105-111
    Number of pages7
    JournalMolecular Oncology
    Volume5
    Issue number1
    DOIs
    Publication statusPublished - 2011
    MoE publication typeA1 Journal article-refereed

    Keywords

    • In situ proximity ligation assay
    • chromosome passenger complex
    • INCENP
    • Aurora B
    • Plk1

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