High-resolution analysis of cell division errors with COIL
The JP group at the University of Edinburgh studies cell division, the essential biological process which ensures genome integrity by equally and identically distributing chromosomes between two daughter cells. Errors in cell division often result in daughter cells with inappropriate chromosome numbers, a condition associated with numerous cancers and birth defects. Although much is known about the basic mechanisms of cell division, structural level mechanistic details of the pathways regulating error-free chromosome segregation are still emerging. In particular, a high-resolution understanding of centromere inheritance and how kinetochores employ dynamic protein interactions to drive chromosome segregation is yet to be obtained.
Dr Maria Alba Abad Fernandez, a postdoc within the JP group, has been studying part of this process – specifically the interaction between the chromosomal passenger complex (CPC) and Shugoshin 1 (Sgo-1), two major regulators of cell division. CPC and Sgo-1 directly interact and need to associate with centromeres to perform their function but the minimal region of Sgo-1 required for their direct binding and how this influences CPC centromere localisation and function were still unknown.
Working with Dr David Kelly within the COIL facility, Dr Abad used cell-based in vivo functional assays and high-resolution microscopy with automated analysis programmes to assess the effect of a number of Sgo-1 mutants on Sgo-1’s ability to bind CPC. Critically, David developed a bespoke ImageJ plugin for the study which enabled Dr Abad to automate the analysis of micrographs and identify the key regions of Sgo-1 required for this interaction in cells. Through this study, Dr Abad discovered that the N-terminus of Sgo-1 is essential for Sgo-1-CPC assembly and that, without this Sgo-1 region, the interaction is disrupted causing perturbed CPC centromere localisation and function, which leads to chromosome missegregation.
The COIL facility houses a range of microscopes capable of performing timelapse imaging at high temporal and spatial resolutions whilst maintaining focus. These can examine specimens from yeast and fungus through to embryos and polymer lattices. The facility staff offer their expertise and guidance to ensure researchers use the correct setup to achieve their desired outcome and are adept at writing bespoke ImageJ plugins to create analysis pipelines. These can assist by speeding up the analysis process or adding functionality to the existing software.
Microscopes capable of performing timelapse imaging at high temporal and spatial resolutions whilst maintaining focus.
Centre for Optical Instrumentation Laboratory
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