Molecular Cell
Volume 77, Issue 1, 2 January 2020, Pages 51-66.e8
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Article
Native Chromatin Proteomics Reveals a Role for Specific Nucleoporins in Heterochromatin Organization and Maintenance

https://doi.org/10.1016/j.molcel.2019.10.018Get rights and content
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Highlights

  • Strategy for affinity purification of native chromatin fragments

  • Heterochromatin and euchromatin have overlapping and non-overlapping proteomes

  • The heterochromatin proteome includes inner nuclear membrane proteins and nucleoporins

  • Inner-ring nucleoporins are required for clustering and maintenance of heterochromatin

Summary

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.

Keywords

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

Cited by (0)

5

These authors contributed equally

6

Present address: Department of Biomedical Engineering, Duke University, Durham, NC, USA

7

Lead Contact