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DNA renaturation activity of the SMC complex implicated in chromosome condensation

Abstract

Chromosome condensation occurs in mitosis before the separation of sister chromatids, and requires DNA topoisomerase II (refs 1,2) and a group of proteins called SMCs3,4,5. The resulting condensed chromosomes in metaphase have a complex hierarchical structure6,7. SMCs, the components of condensed chromosomes, are also required for the separation of sister chromatids and gene dosage compensation, and are found in a range of organisms from yeasts to mammals8,9,10,11,12,13. However, the mechanisms by which the SMCs contribute to chromosome condensation are unknown. We have studied chromosomes in fission-yeast SMC mutants cut3-477 and cut14-208 (ref. 9), which remain largely non-condensed during mitosis at the restrictive temperature (36 °C)9. To test their role in DNA condensation, we isolated the proteins Cut3 and Cut14 as an oligomeric complex, and tested their interactions with isolated DNA. The complex efficiently promoted the DNA renaturation reactions (the winding up of single-strand DNAs into double helical DNA) as much as 70-fold more efficiently than RecA14, which is a bacterial protein with similar activity. The activity of the mutant complex was heat sensitive. As DNA winding by renaturation is a potential cause of supercoiling, the SMC complex may be implicated in promoting the higher-order DNA coiling found in condensed chromosomes.

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Figure 1: Identification of the Cut3–Cut14 complex.
Figure 2: The Cut3–Cut14 complex contains DNA renaturation activity.
Figure 3: Heat-sensitive reannealing activity of the mutant Cut3–Cut14 complex.
Figure 4: S1-sensitive sites in chromatin of cut14 mutant.

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Acknowledgements

We thank T. Horii, H. Ogawa and Y. Adachi for help and advice; T. Hirano for data before publication; and T. Hyman for critically reading the manuscript. This work was supported by grants from the Ministry of Education, Science and Culture of Japan, the Japan Science Technology Foundation (CREST), and the Human Frontier Science Promotion Organization. T.S. is the recipient of a Japan Science Promotion Society fellowship.

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Correspondence to Mitsuhiro Yanagida.

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Sutani, T., Yanagida, . DNA renaturation activity of the SMC complex implicated in chromosome condensation. Nature 388, 798–801 (1997). https://doi.org/10.1038/42062

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