Direct DNA crosslinking with CAP-C uncovers transcription-dependent chromatin organization at high resolution

Nat Biotechnol. 2021 Feb;39(2):225-235. doi: 10.1038/s41587-020-0643-8. Epub 2020 Aug 24.

Abstract

Determining the spatial organization of chromatin in cells mainly relies on crosslinking-based chromosome conformation capture techniques, but resolution and signal-to-noise ratio of these approaches is limited by interference from DNA-bound proteins. Here we introduce chemical-crosslinking assisted proximity capture (CAP-C), a method that uses multifunctional chemical crosslinkers with defined sizes to capture chromatin contacts. CAP-C generates chromatin contact maps at subkilobase (sub-kb) resolution with low background noise. We applied CAP-C to formaldehyde prefixed mouse embryonic stem cells (mESCs) and investigated loop domains (median size of 200 kb) and nonloop domains (median size of 9 kb). Transcription inhibition caused a greater loss of contacts in nonloop domains than loop domains. We uncovered conserved, transcription-state-dependent chromatin compartmentalization at high resolution that is shared from Drosophila to human, and a transcription-initiation-dependent nuclear subcompartment that brings multiple nonloop domains in close proximity. We also showed that CAP-C could be used to detect native chromatin conformation without formaldehyde prefixing.

Publication types

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

MeSH terms

  • Animals
  • CCCTC-Binding Factor / metabolism
  • Chromatin / metabolism*
  • Cross-Linking Reagents / chemistry*
  • DNA (Cytosine-5-)-Methyltransferases / antagonists & inhibitors
  • DNA (Cytosine-5-)-Methyltransferases / metabolism
  • DNA / metabolism*
  • Enzyme Inhibitors / pharmacology
  • Genome
  • Mice
  • Mouse Embryonic Stem Cells / metabolism
  • Nucleic Acid Conformation
  • Promoter Regions, Genetic / genetics
  • Transcription, Genetic*

Substances

  • CCCTC-Binding Factor
  • Chromatin
  • Cross-Linking Reagents
  • Enzyme Inhibitors
  • DNA
  • DNA (Cytosine-5-)-Methyltransferases