Phosphorylated HP1α-Nucleosome Interactions in Phase Separated Environments

J Am Chem Soc. 2023 Nov 8;145(44):23994-24004. doi: 10.1021/jacs.3c06481. Epub 2023 Oct 23.

Abstract

In the nucleus, transcriptionally silent genes are sequestered into heterochromatin compartments comprising nucleosomes decorated with histone H3 Lys9 trimethylation and a protein called HP1α. This protein can form liquid-liquid droplets in vitro and potentially organize heterochromatin through a phase separation mechanism that is promoted by phosphorylation. Elucidating the molecular interactions that drive HP1α phase separation and its consequences on nucleosome structure and dynamics has been challenging due to the viscous and heterogeneous nature of such assemblies. Here, we tackle this problem by a combination of solution and solid-state NMR spectroscopy, which allows us to dissect the interactions of phosphorylated HP1α with nucleosomes in the context of phase separation. Our experiments indicate that phosphorylated human HP1α does not cause any major rearrangements to the nucleosome core, in contrast to the yeast homologue Swi6. Instead, HP1α interacts specifically with the methylated H3 tails and slows the dynamics of the H4 tails. Our results shed light on how phosphorylated HP1α proteins may regulate the heterochromatin landscape, while our approach provides an atomic resolution view of a heterogeneous and dynamic biological system regulated by a complex network of interactions and post-translational modifications.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Chromosomal Proteins, Non-Histone / chemistry
  • Heterochromatin*
  • Histones / chemistry
  • Humans
  • Nucleosomes*
  • Phosphorylation
  • Transcription Factors / metabolism

Substances

  • Nucleosomes
  • Heterochromatin
  • Histones
  • Chromosomal Proteins, Non-Histone
  • Transcription Factors