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Histone H3 K14 acetylation and H2B SUMOylation help regulate binding of the RSC remodeler complex to the nucleosome
Author(s) -
Wilkins Bryan J.,
Jain Neha,
Evans Brian,
Chaudhry Shereen,
Neumann Heinz
Publication year - 2020
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2020.34.s1.00676
Subject(s) - chromatin structure remodeling (rsc) complex , nucleosome , chromatin , histone h3 , microbiology and biotechnology , chemistry , histone , acetylation , sumo protein , chromatin remodeling , biology , dna , biochemistry , ubiquitin , gene
Chromatin remodeler (CR) complexes play crucial roles in regulating chromosomal architectural and act to modify nucleosomal DNA contacts by repositioning nucleosomes. The mechanistic details of the CR family of proteins are of importance because each remodeler complex contributes to unique chromatin structural maintenance. Here, we study the RSC complex of yeast to better characterize its dynamics in the living nucleus. Utilizing an expanded genetic code, we incorporate the photo‐crosslinking amino acid, p‐benzoyl‐L‐phenylalanine (pBPA), site‐specifically into histone proteins in order to monitor non‐histone protein interactions at the nucleosomal surface, in vivo . We map the interface between the RSC motor protein, Sth1, and the nucleosome core particle, revealing that crosslinking efficiency to the H3 N‐terminal tail is dependent upon the acetylation of H3 K14. This modification is also a known binding site of the RSC subunit, Rsc4, suggesting that the remodeler favors binding symmetrical H3 K14ac marks on the same nucleosome. Additionally, we reveal that RSC binding is dependent on SUMOylation of histone H2B. Pairing crosslinking with temporal control of the cell cycle we observe that RSC has differential binding modes as the chromatin architectures alter during compaction and passage into mitosis. All together, this data provides greater mechanistic insight to the functional dynamics of the RSC remodeler under true physiological conditions. Support or Funding Information National Institute Of General Medical Sciences of the National Institutes of Health under [Award Number R15GM124600] German Research Foundation (DFG) [Grants NE1589/5‐1 and 6‐1]