DNA renaturation activity of the SMC complex implicated in chromosome condensation

Publisher Website
Google Scholar
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.