Condensin structures chromosomal DNA through topological links

Abstract

The multisubunit condensin complex is essential for the structural organization of eukaryotic chromosomes during their segregation by the mitotic spindle, but the mechanistic basis for its function is not understood. To address how condensin binds to and structures chromosomes, we have isolated from Saccharomyces cerevisiae cells circular minichromosomes linked to condensin. We find that either linearization of minichromosome DNA or proteolytic opening of the ring-like structure formed through the connection of the two ATPase heads of condensin's structural maintenance of chromosomes (SMC) heterodimer by its kleisin subunit eliminates their association. This suggests that condensin rings encircle chromosomal DNA. We further show that release of condensin from chromosomes by ring opening in dividing cells compromises the partitioning of chromosome regions distal to centromeres. Condensin hence forms topological links within chromatid arms that provide the arms with the structural rigidity necessary for their segregation.

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Figure 1: Linearization releases minichromosome DNA from condensin.
Figure 2: Ring opening by TEV cleavage of Brn1 eliminates condensin function.
Figure 3: Ring opening by Brn1 cleavage releases condensin from minichromosomes.
Figure 4: Ring opening by Brn1 cleavage in vivo releases condensin from chromosomes.
Figure 5: Ring opening by cleavage of Smc4 in vivo releases condensin from chromosomes.
Figure 6: Condensin release by ring opening prevents chromosome arm segregation.
Figure 7: Structuring of chromosomes through topological condensin links.

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Acknowledgements

We thank J. Ellenberg, M. Knop, A.-C. Gavin and all members of the Haering group for advice and discussions, K. Nasmyth (University of Oxford) for strains and plasmids, D. Ivanov for sharing protocols, and the staff of the European Molecular Biology Laboratory (EMBL) Advanced Light Microscopy, Flow Cytometry and Genomics Core facilities for their assistance. This work was supported by funding from EMBL and the German Research Foundation (DFG) Priority Programme 1384 (C.H.H.).

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S.C., J.M. and C.H.H., yeast strain and plasmid generation; S.C., minichromosome, co-immunoprecipitation and ChIP-qPCR experiments; S.C. and J.M., live-cell imaging experiments; C.H.H., chromosome spreads; S.C. and C.H.H., project design and manuscript preparation; C.H.H., project supervision.

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Correspondence to Christian H Haering.

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Supplementary Figures 1–5, Supplementary Tables 1 and 2, and Supplementary Methods (PDF 4948 kb)

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Cuylen, S., Metz, J. & Haering, C. Condensin structures chromosomal DNA through topological links. Nat Struct Mol Biol 18, 894–901 (2011). https://doi.org/10.1038/nsmb.2087

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