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In yeast and metazoa, structural maintenance of chromosome (SMC) complexes play key roles in chromosome segregation, architecture and DNA repair. The main function of the cohesin complex is to hold replicated sister chromatids together until segregation at anaphase, which is dependent on proteolytic cleavage of the cohesin subunit SCC1. Analysis of trypanosomatid genomes showed that the core cohesin and condensin complexes are conserved, but SMC5/6 is absent. To investigate the functional conservation of cohesin in eukaryotes distantly related to yeast and metazoa, we characterized the Trypanosoma brucei SCC1 orthologue. TbSCC1 is expressed prior to DNA synthesis at late G1, remains in the nucleus throughout S- and G2-phases of the cell cycle and disappears at anaphase. Depletion of SCC1 by RNAi or expression of a non-cleavable SCC1 resulted in karyokinesis failure. Using the dominant negative phenotype of non-cleavable SCC1 we investigated checkpoint regulation of cytokinesis in response to mitosis failure at anaphase. In the absence of chromosome segregation, procyclic trypanosomes progressed through cytokinesis to produce one nucleated and one anucleate cell (zoid). In contrast, cytokinesis was incomplete in bloodstream forms, where cleavage was initiated but cells failed to progress to abscission. Kinetoplast duplication was uninterrupted resulting in cells with multiple kinetoplasts and flagella.

Original publication

DOI

10.1111/j.1365-2958.2008.06320.x

Type

Journal article

Journal

Mol Microbiol

Publication Date

08/2008

Volume

69

Pages

666 - 680

Keywords

Animals, Cell Cycle Proteins, Cell Line, Chromosomal Proteins, Non-Histone, Chromosome Segregation, Cytokinesis, Humans, Life Cycle Stages, Molecular Sequence Data, Mutation, Phenotype, Phylogeny, Protein Structure, Tertiary, Protein Subunits, Protozoan Proteins, Sequence Alignment, Trypanosoma brucei brucei