Patterns of histone acetylation in <i>Physarum polycephalum</i>

Waterborg, Jakob H.; Matthews, Harry R.

doi:10.1111/j.1432-1033.1984.tb08290.x

Cited by 50 publications

(20 citation statements)

References 62 publications

(24 reference statements)

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“…In higher animal cells, characteristically, histone H4 is highly acetylated with histone H3 as a close second and low levels of acetylation of H2B and H2A (Matthews and Waterborg, 1985). In P. polycephalum acetylation of transcriptionally active chromatin is limited to acetylation of histones H4 and H3 to high and fairly high levels, respectively (Waterborg and Matthews, 1984). In higher plants, acetylation of histone H3 predominates.…”

Section: Radioactive Acetate Labeling Of Histonesmentioning

confidence: 99%

Histones of Chlamydomonas reinhardtii (Synthesis, Acetylation, and Methylation)

et al. 1995

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Histones of the green alga Chlamydomonas reinhardfii were prepared by a new method and fractionated by reversed-phase high-performance liquid chromatography. Acid-urea-Triton gel analysis and tritiated acetate labeling demonstrated high levels of steady-state acetylation for the single histone H3 protein, in contrast to low levels on histones H4 and H2B. Twenty percent of histone H3 is subject to dynamic acetylation with, on average, three acetylated lysine residues per protein molecule. Histone synthesis in light-dark-synchronized cultures was biphasic with pattern differences between two histone H1 variants, between two H2A variants, and between H2B and ubiquitinated H2B. Automated protein sequence analysis of histone H3 demonstrated a site-specific pattern of steady-state acetylation between 7 and 17% at five of the six amino-terminal lysines and of monomethylation between 5 and 81 O/O at five of the eight amino-terminal lysines in a pattern that may limit dynamic acetylation. An alga1 histone H 3 sequence was confirmed by protein sequencing with a single threonine as residue 28 instead of the serinez8-alaninez9 sequence, present in all other known plant and animal H 3 histones.

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Section: Radioactive Acetate Labeling Of Histonesmentioning

confidence: 99%

Histones of Chlamydomonas reinhardtii (Synthesis, Acetylation, and Methylation)

et al. 1995

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“…uring nucleosome assembly, newly synthesized H4 is acetylated prior to its deposition onto DNA (2,40,71,83). In humans, Drosophila, and Tetrahymena, the acetylation of new H4 takes place in a conserved pattern, at lysines 5 and 12 (the sites are K4 and K11 in Tetrahymena, due to a deletion of the usual arginine residue at position 3) (24,76).…”

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Schizosaccharomyces pombe Hat1 (Kat1) Is Associated with Mis16 and Is Required for Telomeric Silencing

et al. 2012

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The Hat1 histone acetyltransferase has been implicated in the acetylation of histone H4 during chromatin assembly. In this study, we have characterized the Hat1 complex from the fission yeast Schizosaccharomyces pombe and have examined its role in telomeric silencing. Hat1 is found associated with the RbAp46 homologue Mis16, an essential protein. The Hat1 complex acetylates lysines 5 and 12 of histone H4, the sites that are acetylated in newly synthesized H4 in a wide range of eukaryotes. Deletion of hat1 in S. pombe is itself sufficient to cause the loss of silencing at telomeres. This is in contrast to results obtained with an S. cerevisiae hat1⌬ strain, which must also carry mutations of specific acetylatable lysines in the H3 tail domain for loss of telomeric silencing to occur. Notably, deletion of hat1 from S. pombe resulted in an increase of acetylation of histone H4 in subtelomeric chromatin, concomitant with derepression of this region. A similar loss of telomeric silencing was also observed after growing cells in the presence of the deacetylase inhibitor trichostatin A. However, deleting hat1 did not cause loss of silencing at centromeres or the silent mating type locus. These results point to a direct link between Hat1, H4 acetylation, and the establishment of repressed telomeric chromatin in fission yeast. During nucleosome assembly, newly synthesized H4 is acetylated prior to its deposition onto DNA (2,40,71,83). In humans, Drosophila, and Tetrahymena, the acetylation of new H4 takes place in a conserved pattern, at lysines 5 and 12 (the sites are K4 and K11 in Tetrahymena, due to a deletion of the usual arginine residue at position 3) (24, 76). Deacetylation of new H4 occurs over the next 30 to 60 min (40, 73) and is required for proper chromatin maturation (6). The acetylation of new H4 may facilitate the import of H3/H4 dimers into the nucleus (5,15,21,26,31,87). Moreover, recent studies using Physarum as a model system have provided evidence that the K5/K12 diacetylation of H4 is required for efficient nucleosome assembly in that system (26). It therefore seems likely that the rigorous conservation of the diacetylation of nascent H4 reflects an important role in the import/ assembly process.The most likely candidate for the enzyme that acetylates newly synthesized H4 is Hat1 (Kat1), a type B histone acetyltransferase (HAT) (7, 60). Hat1 acetylates free H4 at lysines 5 and 12 in vitro, consistent with the acetylation pattern of new H4 (22,24,46,64,68,75,82). In many organisms Hat1 is associated with p46 (termed Hat2p in Saccharomyces cerevisiae), which stimulates its enzymatic activity (59, 72, 82). Although it was long thought of as predominantly a cytoplasmic enzyme, it is now clear that Hat1 is also present in nuclei (1,51,63,68,72,82). In S. cerevisiae, the nuclear Hat1 complex also contains the protein Hif1p (1, 63).Several lines of evidence have indicated that Hat1 is involved in DNA damage repair (12,14,66) and can be recruited to the sites of DNA double-strand breaks (67). Moreover, in co...

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“…Over the years a method of histone purification from nuclei of Physarum (30,31), alfalfa (32), and Chlamydomonas (33) has been developed that is based on the use of guanidine as a chaotropic salt that prevents protease, deacetylase, and phosphatase action. Histones are obtained quantitatively and at reasonable to high purity.…”

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Steady-state Levels of Histone Acetylation in Saccharomyces cerevisiae

Waterborg

2000

Journal of Biological Chemistry

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The importance of control of the levels of histone acetylation for the control of gene expression in eukaryotic chromatin is being elucidated, and the yeast Saccharomyces cerevisiae has proven to be an important model system. The level of histone acetylation in yeast is the highest known. However, only acetylation of H4 has been quantified, and reports reveal loss of acetylation in histone preparations. A chaotropic guanidine-based method for histone isolation from intact wild-type cells or from a single-step nuclear preparation with butyrate preserves acetylation of all core histones. Histone H4 has an average of more than 2 acetylated lysines per molecule, distributed over 4 sites. Histones H2A, H3, and H2B have 0.2, ϳ2, and >2 acetylated lysines per molecule, respectively, distributed across 2, 5, and 6 sites. Thus, yeast nucleosomes carry, on average, 13 acetylated lysines per octamer, i.e. just above the threshold of 10؊12 deduced for transcriptionally activated chromatin of animals, plants, and algae. Following M r 100,000 ultrafiltration in 2.5% acetic acid, yeast histone H3 was purified to homogeneity by reversed-phase high pressure liquid chromatography. Other core histones were obtained at 80؊95% purity.The yeast Saccharomyces cerevisiae is a unicellular eukaryotic model system used in many studies that aim to define in molecular detail the components of chromatin and the regulatory machinery of gene transcription (1-3). It has been demonstrated in recent years that a localized high level of histone acetylation is required to allow gene transcription to start or continue (4 -9). When histone deacetylases (HDACs) 1 are inhibited by N-butyrate or by specific inhibitors like trichostatin A (TSA), gene transcription tends to increase (10 -13). Conversely, localized recruiting of HDAC activities to methylated DNA results in a repressed state of heterochromatin (9, 14 -17). Currently, analysis of histone acetylation is frequently performed in Western blots of SDS gels using commercial ␣-acetyllysine antibodies (7, 18 -24), but this methodology does not show whether histone acetylation is maintained at levels present in vivo. Commonly, histones, nuclei, or chromatin of yeast are prepared from spheroplasts without the use of HDAC inhibitors (20,(25)(26)(27). Acid-urea (AU) or acid-urea-Triton X-100 (AUT) gels reveal loss of histone H4 and H3 acetylation (8,22,27,28). Proteolysis of histone H3 is observed (19,27).The highest levels of histone acetylation have been reported by Davie et al. (29) when 50Ϫ100 mM butyrate was present during spheroplasting and isolation of nuclei. Histone H4 contained 2.0 Ϯ 0.3 acetylated lysines per molecule, i.e. 10% tetraand 20% triacetylated H4 forms with less than 15% unmodified histone. High levels of acetylation were also shown in twodimensional gels for other core histones, but these were not quantified.Over the years a method of histone purification from nuclei of Physarum (30, 31), alfalfa (32), and Chlamydomonas (33) has been developed that is based on the use of guanid...

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Patterns of histone acetylation in Physarum polycephalum

Cited by 50 publications

References 62 publications

Histones of Chlamydomonas reinhardtii (Synthesis, Acetylation, and Methylation)

Histones of Chlamydomonas reinhardtii (Synthesis, Acetylation, and Methylation)

Schizosaccharomyces pombe Hat1 (Kat1) Is Associated with Mis16 and Is Required for Telomeric Silencing

Steady-state Levels of Histone Acetylation in Saccharomyces cerevisiae

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