The FACT Histone Chaperone Guides Histone H4 Into Its Nucleosomal Conformation in <i>Saccharomyces cerevisiae</i>

McCullough, Laura; Poe, B. S.; Connell, Zaily; Hua, Xin; Formosa, Tim

doi:10.1534/genetics.113.153080

Cited by 10 publications

(14 citation statements)

References 54 publications

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“…The FACT complex is composed of Spt16 and Pob3. FACT makes contacts with the H2A/H2B dimer and has also been shown to interact with histones H3/H4, histone tails, and intact nucleosomes in some contexts (10,(12)(13)(14)(15). Spt16 associates with all five subunits of yeast PAF-C as well as casein kinase II (CKII), as determined by qualitative mass spectrometry analysis (16).…”

Section: Transcription Elongation By Rna Polymerase II (Rnapii)mentioning

confidence: 99%

Quantitative Analysis of Dynamic Protein Interactions during Transcription Reveals a Role for Casein Kinase II in Polymerase-associated Factor (PAF) Complex Phosphorylation and Regulation of Histone H2B Monoubiquitylation

Bedard

Dronamraju

Kerschner

et al. 2016

Journal of Biological Chemistry

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Using affinity purification MS approaches, we have identified a novel role for casein kinase II (CKII) in the modification of the polymerase associated factor complex (PAF-C). Our data indicate that the facilitates chromatin transcription complex (FACT) interacts with CKII and may facilitate PAF complex phosphorylation. Posttranslational modification analysis of affinity-isolated PAF-C shows extensive CKII phosphorylation of all five subunits of PAF-C, although CKII subunits were not detected as interacting partners. Consistent with this, recombinant CKII or FACT-associated CKII isolated from cells can phosphorylate PAF-C in vitro, whereas no intrinsic kinase activity was detected in PAF-C samples. Significantly, PAF-C purifications combined with stable isotope labeling in cells (SILAC) quantitation for PAF-C phosphorylation from wild-type and CKII temperature-sensitive strains (cka1⌬ cka2-8) showed that PAF-C phosphorylation at consensus CKII sites is significantly reduced in cka1⌬ cka2-8 strains. Consistent with a role of CKII in FACT and PAF-C function, we show that decreased CKII function in vivo results in decreased levels of histone H2B lysine 123 monoubiquitylation, a modification dependent on FACT and PAF-C. Taken together, our results define a coordinated role of CKII and FACT in the regulation of RNA polymerase II transcription through chromatin via phosphorylation of PAF-C.

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Section: Transcription Elongation By Rna Polymerase II (Rnapii)mentioning

confidence: 99%

Quantitative Analysis of Dynamic Protein Interactions during Transcription Reveals a Role for Casein Kinase II in Polymerase-associated Factor (PAF) Complex Phosphorylation and Regulation of Histone H2B Monoubiquitylation

Bedard

Dronamraju

Kerschner

et al. 2016

Journal of Biological Chemistry

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“…Genetic analysis of FACT has provided significant insight into the features and physiological functions of nucleosome reorganization (VanDemark et al 2008;McCullough et al 2011McCullough et al , 2013. Mutations in different domains of FACT cause distinct defects in vivo and in vitro, and mutations in histones can either compensate for decreased FACT activity or exacerbate the defects, with different FACT alleles producing different profiles of genetic interactions.…”

mentioning

confidence: 99%

The Abundant Histone Chaperones Spt6 and FACT Collaborate to Assemble, Inspect, and Maintain Chromatin Structure in Saccharomyces cerevisiae

McCullough¹,

Connell²,

Petersen³

et al. 2015

Genetics

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Saccharomyces cerevisiae Spt6 protein is a conserved chromatin factor with several distinct functional domains, including a natively unstructured 30-residue N-terminal region that binds competitively with Spn1 or nucleosomes. To uncover physiological roles of these interactions, we isolated histone mutations that suppress defects caused by weakening Spt6:Spn1 binding with the spt6-F249K mutation. The strongest suppressor was H2A-N39K, which perturbs the point of contact between the two H2A-H2B dimers in an assembled nucleosome. Substantial suppression also was observed when the H2A-H2B interface with H3-H4 was altered, and many members of this class of mutations also suppressed a defect in another essential histone chaperone, FACT. Spt6 is best known as an H3-H4 chaperone, but we found that it binds with similar affinity to H2A-H2B or H3-H4. Like FACT, Spt6 is therefore capable of binding each of the individual components of a nucleosome, but unlike FACT, Spt6 did not produce endonuclease-sensitive reorganized nucleosomes and did not displace H2A-H2B dimers from nucleosomes. Spt6 and FACT therefore have distinct activities, but defects can be suppressed by overlapping histone mutations. We also found that Spt6 and FACT together are nearly as abundant as nucleosomes, with 24,000 Spt6 molecules, 42,000 FACT molecules, and 75,000 nucleosomes per cell. Histone mutations that destabilize interfaces within nucleosomes therefore reveal multiple spatial regions that have both common and distinct roles in the functions of these two essential and abundant histone chaperones. We discuss these observations in terms of different potential roles for chaperones in both promoting the assembly of nucleosomes and monitoring their quality.

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“…These chaperones incorporate canonical H3.1/H4 into DNA during replication, and ASF1 also collaborates with HIRA to exchange H3.1 for H3.3 at transcriptionally active sites. Processes of H3/H4 histone loading require a specific conformation that is facilitated by histone chaperones FACT facilitates chromatin transcription 1 or NAP1 nucleosome assembly protein 1 (Bowman et al ., ; McCullough et al ., ). These factors are primarily involved in deposition of the other core histone components (H2A/H2B dimers) (Ishimi et al ., ; Ito et al ., ; Hondele et al ., ).…”

Section: Modulation Of Telomere and Rdna Chromatin By Histone Variantmentioning

confidence: 97%

Chromatin dynamics of plant telomeres and ribosomal genes

2015

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SUMMARYTelomeres and genes encoding 45S ribosomal RNA (rDNA) are frequently located adjacent to each other on eukaryotic chromosomes. Although their primary roles are different, they show striking similarities with respect to their features and additional functions. Both genome domains have remarkably dynamic chromatin structures. Both are hypersensitive to dysfunctional histone chaperones, responding at the genomic and epigenomic levels. Both generate non-coding transcripts that, in addition to their epigenetic roles, may induce gross chromosomal rearrangements. Both give rise to chromosomal fragile sites, as their replication is intrinsically problematic. However, at the same time, both are essential for maintenance of genomic stability and integrity. Here we discuss the structural and functional inter-connectivity of telomeres and rDNA, with a focus on recent results obtained in plants.

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

Cited by 10 publications

References 54 publications

Quantitative Analysis of Dynamic Protein Interactions during Transcription Reveals a Role for Casein Kinase II in Polymerase-associated Factor (PAF) Complex Phosphorylation and Regulation of Histone H2B Monoubiquitylation

Quantitative Analysis of Dynamic Protein Interactions during Transcription Reveals a Role for Casein Kinase II in Polymerase-associated Factor (PAF) Complex Phosphorylation and Regulation of Histone H2B Monoubiquitylation

The Abundant Histone Chaperones Spt6 and FACT Collaborate to Assemble, Inspect, and Maintain Chromatin Structure in Saccharomyces cerevisiae

Chromatin dynamics of plant telomeres and ribosomal genes

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