In Vitro Kinetochore Assembly.
The kinetochore is the primary site of interaction between chromosomes and microtubules of the mitotic spindle during chromosome segregation. Kinetochores are composed of more than 100 proteins that transiently assemble during mitosis at a single epigenetically defined region on each chromosome, known as the centromere. Kinetochore assembly and activity must be tightly regulated to ensure proper microtubule interaction and faithful chromosome segregation. Kinetochore malfunction can result in chromosome segregation defects leading to aneuploidy and cell death. As such, cell free and reconstituted systems to analyze kinetochore formation and function are invaluable in probing the biochemical activities of kinetochores. In vitro approaches to studying kinetochores have enabled the manipulation of kinetochore protein structure, function, interactions, and regulation that are not possible in cells. Here we outline a cell-free approach for the assembly of centromeres and recruitment of functional kinetochores that enables their manipulation and analysis.
['Miell|Matthew D D|MD|', 'Straight|Aaron F|AF|']
D000818:Animals D059447:Cell Cycle Checkpoints D002457:Cell Extracts D002503:Centromere D000076247:Centromere Protein A D002843:Chromatin D005260:Female D015870:Gene Expression D006657:Histones D018386:Kinetochores D008677:Metaphase D008870:Microtubules D008938:Mitosis D009707:Nucleosomes D009865:Oocytes D055503:Protein Multimerization D008941:Spindle Apparatus D014981:Xenopus
Kinetochore Centromere Chromatin In vitro assembly Xenopus egg extract