The Carboxyl Terminus of Rtt109 Functions in Chaperone Control of Histone Acetylation

Radovani, Ernest; Cadorin, Matthew D. R.; Shams, Tahireh A.; El‐Rass, Suzan; Karsou, Abdel Rahman; Kim, Hyun Soo; Kurat, Christoph F.; Keogh, Michael‐Christopher; Greenblatt, Jack; Fillingham, Jeffrey

doi:10.1128/ec.00291-12

Cited by 18 publications

(29 citation statements)

References 67 publications

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“…Rtt109 was originally identified for its ability to support the acetylation of H3K56 [ 2 , 42 , 43 ], but our data indicate that this is not its primary or initial site of acetylation under our experimental conditions. We identified one possible difference: the use of histones purified from chicken erythrocytes [ 44 ]. We therefore wanted to determine if these different histone substrates combined with longer incubation time are key to facilitating H3K56 acetylation.…”

Section: Resultsmentioning

confidence: 99%

Utilizing Targeted Mass Spectrometry to Demonstrate Asf1-Dependent Increases in Residue Specificity for Rtt109-Vps75 Mediated Histone Acetylation

et al. 2015

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In Saccharomyces cerevisiae, Rtt109, a lysine acetyltransferase (KAT), associates with a histone chaperone, either Vps75 or Asf1. It has been proposed that these chaperones alter the selectivity of Rtt109 or which residues it preferentially acetylates. In the present study, we utilized a label-free quantitative mass spectrometry-based method to determine the steady-state kinetic parameters of acetylation catalyzed by Rtt109-Vps75 on H3 monomer, H3/H4 tetramer, and H3/H4-Asf1 complex. These results show that among these histone conformations, only H3K9 and H3K23 are significantly acetylated under steady-state conditions and that Asf1 promotes H3/H4 acetylation by Rtt109-Vps75. Asf1 equally increases the Rtt109-Vps75 specificity for both of these residues with a maximum stoichiometry of 1:1 (Asf1 to H3/H4), but does not alter the selectivity between these two residues. These data suggest that the H3/H4-Asf1 complex is a substrate for Rtt109-Vps75 without altering selectivity between residues. The deletion of either Rtt109 or Asf1 in vivo results in the same reduction of H3K9 acetylation, suggesting that Asf1 is required for efficient H3K9 acetylation both in vitro and in vivo. Furthermore, we found that the acetylation preference of Rtt109-Vps75 could be directed to H3K56 when those histones already possess modifications, such as those found on histones purified from chicken erythrocytes. Taken together, Vps75 and Asf1 both enhance Rtt109 acetylation for H3/H4, although via different mechanisms, but have little impact on the residue selectivity. Importantly, these results provide evidence that histone chaperones can work together via interactions with either the enzyme or the substrate to more efficiently acetylate histones.

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Section: Resultsmentioning

confidence: 99%

Utilizing Targeted Mass Spectrometry to Demonstrate Asf1-Dependent Increases in Residue Specificity for Rtt109-Vps75 Mediated Histone Acetylation

et al. 2015

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“…While Rtt109 catalyzes H3K56ac via the Asf1-H3-H4 substrate in vivo , the role of Vps75 appears to relate more to Rtt109-catalyzed acetylation of the H3 tail. Recently, a study involving the C-terminal carboxy domain of Rtt109 has provided evidence that Vps75 may play some role in promoting H3K56ac in vivo (Radovani et al , 2013), observations that demonstrate the complex interactions between Rtt109, histone chaperones and histone substrates.…”

Section: Dna Replication-coupled Nucleosome Assemblymentioning

confidence: 99%

“…Rtt109 and its complexes do not significantly acetylate histone H2A, H2B, or H4 in vitro (Tsubota et al , 2007). In terms of other in vitro substrates, the Rtt109-Vps75 complex is capable of acetylating histone H1 linker under some conditions in vitro , though the functional significance of this ability is unknown (Radovani et al , 2013). …”

Section: A Closer Look At Rtt109 Hats: Molecular Biochemical and Strmentioning

confidence: 99%

Histone-modifying enzymes, histone modifications and histone chaperones in nucleosome assembly: Lessons learned from Rtt109 histone acetyltransferases

Dahlin

Chen

Walters

et al. 2014

Critical Reviews in Biochemistry and Molecular Biology

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During DNA replication, nucleosomes ahead of replication forks are disassembled to accommodate replication machinery. Following DNA replication, nucleosomes are then reassembled onto replicated DNA using both parental and newly synthesized histones. This process, termed DNA replication-coupled nucleosome assembly (RCNA), is critical for maintaining genome integrity and for the propagation of epigenetic information, dysfunctions of which have been implicated in cancers and aging. In recent years, it has been shown that RCNA is carefully orchestrated by a series of histone modifications, histone chaperones and histone-modifying enzymes. Interestingly, many features of RCNA are also found in processes involving DNA replication-independent nucleosome assembly like histone exchange and gene transcription. In yeast, histone H3 lysine K56 acetylation (H3K56ac) is found in newly synthesized histone H3 and is critical for proper nucleosome assembly and for maintaining genomic stability. The histone acetyltransferase (HAT) regulator of Ty1 transposition 109 (Rtt109) is the sole enzyme responsible for H3K56ac in yeast. Much research has centered on this particular histone modification and histone-modifying enzyme. This Critical Review summarizes much of our current understanding of nucleosome assembly and highlights many important insights learned from studying Rtt109 HATs in fungi. We highlight some seminal features in nucleosome assembly conserved in mammalian systems and describe some of the lingering questions in the field. Further studying fungal and mammalian chromatin assembly may have important public health implications, including deeper understandings of human cancers and aging as well as the pursuit of novel anti-fungal therapies.

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“…In yeast, Rtt109 is an acetyltransferase that modifies various H3-H4 lysine residues, including H3 K9, K27, and K56 ( Schneider et al, 2006 ; Fillingham et al, 2008 ; Radovani et al, 2013 ). H3-K56 acetylation is important in fungal pathogenicity, making Rtt109 an attractive antifungal therapeutic target ( Wurtele et al, 2010 ; Dahlin et al, 2014 ).…”

Section: Introductionmentioning

confidence: 99%

Stoichiometry of Rtt109 complexes with Vps75 and histones H3-H4

et al. 2020

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Histone acetylation is one of many posttranslational modifications that affect nucleosome accessibility. Vps75 is a histone chaperone that stimulates Rtt109 acetyltransferase activity toward histones H3-H4 in yeast. In this study, we use sedimentation velocity and light scattering to characterize various Vps75–Rtt109 complexes, both with and without H3-H4. These complexes were previously ill-defined because of protein multivalency and oligomerization. We determine both relative and absolute stoichiometry and define the most pertinent and homogeneous complexes. We show that the Vps75 dimer contains two unequal binding sites for Rtt109, with the weaker binding site being dispensable for H3-H4 acetylation. We further show that the Vps75–Rtt109–(H3-H4) complex is in equilibrium between a 2:1:1 species and a 4:2:2 species. Using a dimerization mutant of H3, we show that this equilibrium is mediated by the four-helix bundle between the two copies of H3. We optimize the purity, yield, and homogeneity of Vps75–Rtt109 complexes and determine optimal conditions for solubility when H3-H4 is added. Our comprehensive biochemical and biophysical approach ultimately defines the large-scale preparation of Vps75–Rtt109–(H3-H4) complexes with precise stoichiometry. This is an essential prerequisite for ongoing high-resolution structural and functional analysis of this important multi-subunit complex.

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The Carboxyl Terminus of Rtt109 Functions in Chaperone Control of Histone Acetylation

Cited by 18 publications

References 67 publications

Utilizing Targeted Mass Spectrometry to Demonstrate Asf1-Dependent Increases in Residue Specificity for Rtt109-Vps75 Mediated Histone Acetylation

Utilizing Targeted Mass Spectrometry to Demonstrate Asf1-Dependent Increases in Residue Specificity for Rtt109-Vps75 Mediated Histone Acetylation

Histone-modifying enzymes, histone modifications and histone chaperones in nucleosome assembly: Lessons learned from Rtt109 histone acetyltransferases

Stoichiometry of Rtt109 complexes with Vps75 and histones H3-H4

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