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 journalArticleScientificpeer-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

Fingerprint

Ligation
Centromere
Mitosis
Phosphotransferases
Maintenance
Anaphase
Genomic Instability
Cell Division
Proteins
Antibodies

Keywords

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

Cite this

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title = "Spatio-temporal composition of the mitotic Chromosomal Passenger Complex detected using in situ proximity ligation assay: Technical Note",
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.",
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Spatio-temporal composition of the mitotic Chromosomal Passenger Complex detected using in situ proximity ligation assay : Technical Note. / Vuoriluoto, Mariaana; Laine, Leena; Saviranta, Petri; Pouwels, Jeroen; Kallio, Marko.

In: Molecular Oncology, Vol. 5, No. 1, 2011, p. 105-111.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Spatio-temporal composition of the mitotic Chromosomal Passenger Complex detected using in situ proximity ligation assay

T2 - Technical Note

AU - Vuoriluoto, Mariaana

AU - Laine, Leena

AU - Saviranta, Petri

AU - Pouwels, Jeroen

AU - Kallio, Marko

PY - 2011

Y1 - 2011

N2 - 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.

AB - 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.

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