Native Chromatin Proteomics Reveals a Role for Specific Nucleoporins in Heterochromatin Organization and Maintenance

Nahid Iglesias, Joao A. Paulo, Antonis Tatarakis, Xiaoyi Wang, Amanda L. Edwards, Natarajan V. Bhanu, Benjamin A. Garcia, Wilhelm Haas, Steven P. Gygi, Danesh Moazed

Research output: Contribution to journalArticlepeer-review

31 Scopus citations

Abstract

Spatially and functionally distinct domains of heterochromatin and euchromatin play important roles in the maintenance of chromosome stability and regulation of gene expression, but a comprehensive knowledge of their composition is lacking. Here, we develop a strategy for the isolation of native Schizosaccharomyces pombe heterochromatin and euchromatin fragments and analyze their composition by using quantitative mass spectrometry. The shared and euchromatin-specific proteomes contain proteins involved in DNA and chromatin metabolism and in transcription, respectively. The heterochromatin-specific proteome includes all proteins with known roles in heterochromatin formation and, in addition, is enriched for subsets of nucleoporins and inner nuclear membrane (INM) proteins, which associate with different chromatin domains. While the INM proteins are required for the integrity of the nucleolus, containing ribosomal DNA repeats, the nucleoporins are required for aggregation of heterochromatic foci and epigenetic inheritance. The results provide a comprehensive picture of heterochromatin-associated proteins and suggest a role for specific nucleoporins in heterochromatin function.

Original languageEnglish
Pages (from-to)51-66.e8
JournalMolecular cell
Volume77
Issue number1
DOIs
StatePublished - Jan 2 2020

Keywords

  • Bdf1
  • Clr4
  • H3K9me
  • HP1
  • INM proteins
  • NPC
  • SUV39H
  • Swi6
  • epigenetic inheritance
  • nucleoporins

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