SIR proteins create compact heterochromatin fibers

Sarah G. Swygert, Subhadip Senapati, Mehmet F. Bolukbasi, Scot A. Wolfe, Stuart Lindsay, Craig L. Peterson

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Heterochromatin is a silenced chromatin region essential for maintaining genomic stability and driving developmental processes. The complicated structure and dynamics of heterochromatin have rendered it difficult to characterize. In budding yeast, heterochromatin assembly requires the SIR proteins—Sir3, believed to be the primary structural component of SIR heterochromatin, and the Sir2–4 complex, responsible for the targeted recruitment of SIR proteins and the deacetylation of lysine 16 of histone H4. Previously, we found that Sir3 binds but does not compact nucleosomal arrays. Here we reconstitute chromatin fibers with the complete complement of SIR proteins and use sedimentation velocity, molecular modeling, and atomic force microscopy to characterize the stoichiometry and conformation of SIR chromatin fibers. In contrast to fibers with Sir3 alone, our results demonstrate that SIR arrays are highly compact. Strikingly, the condensed structure of SIR heterochromatin fibers requires both the integrity of H4K16 and an interaction between Sir3 and Sir4. We propose a model in which a dimer of Sir3 bridges and stabilizes two adjacent nucleosomes, while a Sir2–4 heterotetramer interacts with Sir3 associated with a nucleosomal trimer, driving fiber compaction.

Original languageEnglish (US)
Pages (from-to)12447-12452
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume115
Issue number49
DOIs
StatePublished - Dec 4 2018
Externally publishedYes

Fingerprint

Heterochromatin
Chromatin
Proteins
Saccharomycetales
Nucleosomes
Atomic Force Microscopy
Genomic Instability
Histones
Lysine
Complement System Proteins

Keywords

  • Chromatin
  • Heterochromatin
  • Sedimentation
  • Sir

ASJC Scopus subject areas

  • General

Cite this

SIR proteins create compact heterochromatin fibers. / Swygert, Sarah G.; Senapati, Subhadip; Bolukbasi, Mehmet F.; Wolfe, Scot A.; Lindsay, Stuart; Peterson, Craig L.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 115, No. 49, 04.12.2018, p. 12447-12452.

Research output: Contribution to journalArticle

Swygert, Sarah G. ; Senapati, Subhadip ; Bolukbasi, Mehmet F. ; Wolfe, Scot A. ; Lindsay, Stuart ; Peterson, Craig L. / SIR proteins create compact heterochromatin fibers. In: Proceedings of the National Academy of Sciences of the United States of America. 2018 ; Vol. 115, No. 49. pp. 12447-12452.
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