Unknown

Dataset Information

0

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


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.

SUBMITTER: Vuoriluoto M 

PROVIDER: S-EPMC5528271 | BioStudies | 2011-01-01

SECONDARY ACCESSION(S): 10.1016/j.molonc.2010.10.002

REPOSITORIES: biostudies

Similar Datasets

1000-01-01 | S-EPMC1369225 | BioStudies
2009-01-01 | S-EPMC3442196 | BioStudies
2004-01-01 | S-EPMC2172304 | BioStudies
2020-01-01 | S-EPMC7520452 | BioStudies
2017-01-01 | S-EPMC5379950 | BioStudies
1000-01-01 | S-EPMC2613119 | BioStudies
2015-01-01 | S-EPMC4392389 | BioStudies
2019-01-01 | S-EPMC6891087 | BioStudies
2009-01-01 | S-EPMC2655253 | BioStudies
2020-01-01 | S-EPMC6992762 | BioStudies