Detecting chromatin interactions between and along sister chromatids with SisterC

Nat Methods. 2020 Oct;17(10):1002-1009. doi: 10.1038/s41592-020-0930-9. Epub 2020 Sep 23.

Abstract

Chromosome segregation requires both compaction and disentanglement of sister chromatids. We describe SisterC, a chromosome conformation capture assay that distinguishes interactions between and along identical sister chromatids. SisterC employs 5-bromo-2'-deoxyuridine (BrdU) incorporation during S-phase to label newly replicated strands, followed by Hi-C and then the destruction of 5-bromodeoxyuridine-containing strands via Hoechst/ultraviolet treatment. After sequencing of the remaining intact strands, this allows assignment of Hi-C products as inter- and intra-sister interactions based on the strands that reads are mapped to. We performed SisterC on mitotic Saccharomyces cerevisiae cells. We find precise alignment of sister chromatids at centromeres. Along arms, sister chromatids are less precisely aligned, with inter-sister connections every ~35 kilobase (kb). Inter-sister interactions occur between cohesin binding sites that are often offset by 5 to 25 kb. Along sister chromatids, cohesin results in the formation of loops of up to 50 kb. SisterC allows study of the complex interplay between sister chromatid compaction and their segregation during mitosis.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Chromatids / physiology*
  • Chromatin / physiology*
  • Chromosome Segregation / physiology*
  • DNA Repair
  • DNA Replication
  • Gene Expression Regulation
  • Mitosis / physiology
  • Saccharomyces cerevisiae / metabolism
  • Saccharomyces cerevisiae / physiology

Substances

  • Chromatin