Structural basis of histone H2A–H2B recognition by the essential chaperone FACT

Hondele, Maria; Stuwe, T.; Hassler, Markus; Halbach, Felix; Bowman, Andrew; Zhang, Elisa T.; Nijmeijer, Bianca; Kotthoff, Christiane; Rybin, Vladimir; Amlacher, Stefan; Hurt, Ed; Ladurner, Andreas G.

doi:10.1038/nature12242

Cited by 166 publications

(210 citation statements)

References 41 publications

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“…Nhp6 constitutes an initial step in FACT-dependent nucleosome reorganization that involves DNA bending and altered histone-DNA contacts to allow FACT to substantially change nucleosome structure (Xin et al 2009). Recent data suggest that DNA topology at the nucleosomal entry/exit site might influence FACT activity on nucleosomes (Hondele et al 2013). Reconstituted CENP-A Cse4 nucleosomes have weakened DNA-histone contacts at the DNA entry/exit site compared with H3 nucleosomes (Dechassa et al 2011;Tachiwana et al 2011), which may generate an open chromatin configuration that allows FACT to function on CENP-A Cse4 nucleosomes independent of Nhp6.…”

Section: Discussionmentioning

confidence: 99%

The FACT complex interacts with the E3 ubiquitin ligase Psh1 to prevent ectopic localization of CENP-A

Deyter

Biggins

2014

Genes Dev.

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Centromere identity and its epigenetic maintenance require the incorporation of a histone H3 variant called CENP-A at centromeres. CENP-A mislocalization to ectopic sites may disrupt chromatin-based processes and chromosome segregation, so it is important to uncover the mechanisms by which this variant is exclusively localized to centromeres. Here, we identify a role for the conserved chromatin-modifying complex FACT (facilitates chromatin transcription/transactions) in preventing budding yeast CENP-A Cse4 mislocalization to euchromatin by mediating its proteolysis. The Spt16 subunit of the FACT complex binds to Psh1 (Pob3/Spt16/ histone), an E3 ubiquitin ligase that targets CENP-A Cse4 for degradation. The interaction between Psh1 and Spt16 is critical for both CENP-A Cse4 ubiquitylation and its exclusion from euchromatin. We found that Psh1 cannot efficiently ubiquitylate CENP-A Cse4 nucleosomes in vitro, suggesting that additional factors must facilitate CENP-A Cse4 removal from chromatin in vivo. Consistent with this, a Psh1 mutant that cannot associate with FACT has a reduced interaction with CENP-A Cse4 in vivo. Together, our data identify a previously unknown mechanism to maintain centromere identity and genomic stability through the FACT-mediated degradation of ectopically localized CENP-A Cse4 .

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Section: Discussionmentioning

confidence: 99%

The FACT complex interacts with the E3 ubiquitin ligase Psh1 to prevent ectopic localization of CENP-A

Deyter

Biggins

2014

Genes Dev.

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“…In vivo , the assembly process is catalyzed by histone chaperones, a diverse class of proteins that is involved in histone turnover including transport, transfer, and storage (De Koning et al , 2007). Structural studies of histone chaperones bound to cognate histones have revealed that histone chaperones frequently protect hydrophobic histone interfaces that become solvent‐exposed outside of the context of the nucleosome or neutralize excess positive charge by providing an acidic binding pocket to directly compete with non‐specific electrostatic histone–DNA interactions (English et al , 2006; Zhou et al , 2008, 2011; Cho & Harrison, 2011; Hu et al , 2011; Elsasser et al , 2012; Liu et al , 2012; Hondele et al , 2013; Obri et al , 2014; Huang et al , 2015). However, how chaperones mediate the final deposition onto DNA to assemble nucleosomes remains poorly understood (Loyola & Almouzni, 2004; De Koning et al , 2007; Elsasser & D'Arcy, 2012).…”

Section: Introductionmentioning

confidence: 99%

Structural evidence for Nap1‐dependent H2A–H2B deposition and nucleosome assembly

et al. 2016

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Nap1 is a histone chaperone involved in the nuclear import of H2A–H2B and nucleosome assembly. Here, we report the crystal structure of Nap1 bound to H2A–H2B together with in vitro and in vivo functional studies that elucidate the principles underlying Nap1‐mediated H2A–H2B chaperoning and nucleosome assembly. A Nap1 dimer provides an acidic binding surface and asymmetrically engages a single H2A–H2B heterodimer. Oligomerization of the Nap1–H2A–H2B complex results in burial of surfaces required for deposition of H2A–H2B into nucleosomes. Chromatin immunoprecipitation‐exonuclease (ChIP‐exo) analysis shows that Nap1 is required for H2A–H2B deposition across the genome. Mutants that interfere with Nap1 oligomerization exhibit severe nucleosome assembly defects showing that oligomerization is essential for the chaperone function. These findings establish the molecular basis for Nap1‐mediated H2A–H2B deposition and nucleosome assembly.

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“…FACT was initially shown to be an H2A-H2B chaperone (Orphanides et al 1999;Belotserkovskaya et al 2003), but it also binds to H3-H4, intact nucleosomes, the N-terminal tails of some histones, and DNA (DNA binding is through the HMGB domain of SSRP1 or the separate Nhp6 protein in yeast) (Orphanides et al 1999;Formosa et al 2001;Stuwe et al 2008;Kemble et al 2013). Structures have been determined for several individual domains of FACT (Allain et al 1999;VanDemark et al 2006;Stuwe et al 2008;VanDemark et al 2008;Hondele et al 2013;Kemble et al 2013) and each of these domains contributes to the interactions with nucleosomal components, suggesting that FACT can bind to the different components simultaneously through independent contacts. FACT also binds directly to several transcription and replication factors (Wittmeyer et al 1999;Squazzo et al 2002;Simic et al 2003;Takahata et al 2009a;Formosa 2012).…”

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confidence: 99%

The FACT Histone Chaperone Guides Histone H4 Into Its Nucleosomal Conformation in Saccharomyces cerevisiae

et al. 2013

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The pob3-Q308K mutation alters the small subunit of the Saccharomyces cerevisiae histone/nucleosome chaperone Facilitates Chromatin Transactions (FACT), causing defects in both transcription and DNA replication. We describe histone mutations that suppress some of these defects, providing new insight into the mechanism of FACT activity in vivo. FACT is primarily known for its ability to promote reorganization of nucleosomes into a more open form, but neither the pob3-Q308K mutation nor the compensating histone mutations affect this activity. Instead, purified mutant FACT complexes fail to release from nucleosomes efficiently, and the histone mutations correct this flaw. We confirm that pob3-T252E also suppresses pob3-Q308K and show that combining two suppressor mutations can be detrimental, further demonstrating the importance of balance between association and dissociation for efficient FACT:nucleosome interactions. To explain our results, we propose that histone H4 can adopt multiple conformations, most of which are incompatible with nucleosome assembly. FACT guides H4 to adopt appropriate conformations, and this activity can be enhanced or diminished by mutations in Pob3 or histones. FACT can therefore destabilize nucleosomes by favoring the reorganized state, but it can also promote assembly by tethering histones and DNA together and maintaining them in conformations that promote canonical nucleosome formation.

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Structural basis of histone H2A–H2B recognition by the essential chaperone FACT

Cited by 166 publications

References 41 publications

The FACT complex interacts with the E3 ubiquitin ligase Psh1 to prevent ectopic localization of CENP-A

The FACT complex interacts with the E3 ubiquitin ligase Psh1 to prevent ectopic localization of CENP-A

Structural evidence for Nap1‐dependent H2A–H2B deposition and nucleosome assembly

The FACT Histone Chaperone Guides Histone H4 Into Its Nucleosomal Conformation in Saccharomyces cerevisiae

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