Antibodies specific to acetylated histones document the existence of deposition- and transcription-related histone acetylation in Tetrahymena.

Lin, Ren-Yong; Leone, Joseph W.; Cook, Richard G.; Allis, C. David

doi:10.1083/jcb.108.5.1577

Cited by 139 publications

(104 citation statements)

References 30 publications

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“…Of the histone H2A synthesized, a limited amount moves into the nucleus and is acetylated (26), and the excess unacetylated histone H2A accumulates in the cytoplasm, eventually secreted into the lumen. While the functional role of acetylation is poorly understood, it is thought to play a role in the formation of either newly synthesized or transcriptionally competent chromatin (27)(28)(29). We also observed the immunoreactivity to unacetylated histone H2A in the endoplasmic reticulum of some gastric gland cells, which would be wholly consistent with de novo histone synthesis in the cytoplasm.…”

Section: Discussionsupporting

confidence: 56%

Pepsin-Mediated Processing of the Cytoplasmic Histone H2A to Strong Antimicrobial Peptide Buforin I

Kim

Yoon

Minn

et al. 2000

The Journal of Immunology

110

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The intestinal epithelium forms a first line of innate host defense by secretion of proteins with antimicrobial activity against microbial infection. Despite the extensive studies on the antimicrobial host defense in many gastrointestinal tracts, little is known about the antimicrobial defense system of the stomach. The potent antimicrobial peptide buforin I, consisting of 39 aa, was isolated recently from the stomach tissue of an Asian toad, Bufo bufo gargarizans. In this study we examined the mechanism of buforin I production in toad stomach tissue. Buforin I is produced by the action of pepsin isozymes, named pepsin Ca and Cb, cleaving the Tyr39-Ala40 bond of histone H2A. Immunohistochemical analysis revealed that buforin I is present extracellularly on the mucosal surface, and unacetylated histone H2A, a precursor of buforin I, is localized in the cytoplasm of gastric gland cells. Furthermore, Western blot analysis showed that buforin I is also present in the gastric fluids, and immunoelectron microscopy detected localization of the unacetylated histone H2A in the cytoplasmic granules of gastric gland cells. The distinct subcellular distribution of the unacetylated histone H2A and the detection of the unacetylated buforin I both on the mucosal surface and in the lumen suggest that buforin I is produced from the cytoplasmic unacetylated histone H2A secreted into the gastric lumen and subsequently processed by pepsins. Our results indicate that buforin I along with pepsins in the vertebrate stomach may contribute to the innate host defense of the stomach against invading microorganisms.

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Section: Discussionsupporting

confidence: 56%

Pepsin-Mediated Processing of the Cytoplasmic Histone H2A to Strong Antimicrobial Peptide Buforin I

Kim

Yoon

Minn

et al. 2000

The Journal of Immunology

110

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“…This antibody, prepared as described previously (14), was raised against the first 20 amino acids of tetraacetylated histone H4 from Tetrahymena, which differs significantly from that of Xenopus. To determine whether this antibody shows the same specificity for hyperacetylated Xenopus histone H4, a Western blot analysis was performed using histones from butyrate-treated HeLa cells (X. laevis histone H4 shares the identical first 20 amino acids with human H4).…”

Section: Resultsmentioning

confidence: 99%

“…Western Blot Analysis-A polyclonal antibody specific for acetylated histone H4 was prepared as described previously (14). Western blot analysis (15) was performed with histones isolated from HeLa cells grown in the presence of sodium butyrate (16 (17).…”

Section: Methodsmentioning

confidence: 99%

“…The membrane was alkaline fixed, blocked, and hybridized, as per manufacturer's instructions to a 180 bp, [␣-32 P]dATP end-labeled, HindIII/EcoRI fragment from plasmid pXlo (13) that contains a copy of the X. laevis oocyte 5 S rRNA gene coding sequence. In vitro transcribed X. laevis oocyte and somatic 5 S rRNAs were prepared by T7 RNA polymerase (New England Biolabs, Beverly, MA) transcription of DraI-digested plasmids pXlo and pXls, respectively (13), as per the manufacturer's instructions.Western Blot Analysis-A polyclonal antibody specific for acetylated histone H4 was prepared as described previously (14). Western blot analysis (15) was performed with histones isolated from HeLa cells grown in the presence of sodium butyrate (16).…”

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confidence: 99%

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Transcriptionally Active Xenopus laevis Somatic 5 S Ribosomal RNA Genes Are Packaged with Hyperacetylated Histone H4, Whereas Transcriptionally Silent Oocyte Genes Are Not

Howe¹,

Ranalli²,

Allis³

et al. 1998

Journal of Biological Chemistry

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The relationship between histone acetylation and transcription of the Xenopus laevis oocyte and somatic 5 S ribosomal RNA genes was investigated. Chromatin fragments from a X. laevis kidney cell line were immunoprecipitated with an antibody specific for hyperacetylated histone H4. The DNA from the hyperacetylated chromatin was probed with both oocyte-and somatic gene-specific sequences, and the results showed that the upstream, nontranscribed region of the transcriptionally active somatic genes is packaged with acetylated histone H4. In contrast, the corresponding region of the transcriptionally silent oocyte genes is packaged with hypoacetylated histone H4 in this cells line. Further study also showed that this region of the oocyte genes was less sensitive to digestion with the enzyme, micrococcal nuclease. Together these results suggest that, as described for both RNA polymerase I and II transcribed genes, there is a correlation between histone acetylation and transcription of the RNA polymerase III transcribed 5 S ribosomal RNA genes in X. laevis.The acetylation of lysine residues within the amino-terminal "tails" of core histones is known to be associated with transcriptionally active genes, although the basis of this relationship is not well understood (for reviews see Refs. 1-4). The use of antibodies, specific for hyperacetylated histones, to map core histone acetylation of active and inactive genes has suggested that acetylation is not a consequence but rather a precondition of transcription. It has been demonstrated in chicken embryonic erythrocytes, for example, that the ␤ globin genes are packaged with acetylated histones prior to, during, and after activation during development (5). Furthermore, the plateletderived growth factor B chain gene is also found packaged with hyperacetylated histones prior to gene induction in a human hematopoietic stem cell line (6). In contrast to this apparent constitutive presence of acetylated histones on "poised" RNA polymerase II transcribed genes, transcriptionally silent 28 S rRNA 1 genes in rat tissue culture cells are packaged with nonacetylated histones (7). This latter result is despite the fact that the same study showed these silent genes exhibited increased sensitivity to micrococcal nuclease, another hallmark of transcriptionally poised genes.Xenopus laevis produces two major types of 5 S rRNA: the somatic type, which is transcribed in most cells, and the oocyte type, which is produced only during early oogenesis, early embryogenesis, and in certain tissue culture cell lines (8, 9). Each 5 S rRNA type is transcribed from a distinct multigene family, and considerable research has focused on explaining the differential expression of these genes in somatic cells. Recently, it has been reported that histone acetylation enhances RNA polymerase III transcription of dinucleosomal 5 S rRNA gene templates (10), suggesting a possible role for histone acetylation in 5 S rRNA transcription. It was also demonstrated that in a X. laevis kidney cell line transcribing low ...

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“…A potential mechanism for the function of Sir2p in silencing has been suggested by the observation that SIR2 overexpression is correlated with hypoacetylation of a subset of histones (Braunstein et al, 1993). Hypoacetylated histones are often found at silenced or inactive loci in yeast and other organisms (Lin et al, 1989;Turner et al, 1992;Braunstein et al, 1993Braunstein et al, , 1996Jeppesen and Turner, 1993;O'Neill and Turner, 1995). However, SIR2's potential role in modulating histone deacetylation may be indirect as histone deacetylase activity has not been detected for Sir2p in vitro (Braunstein et al, 1993).…”

Section: Introductionmentioning

confidence: 99%

The Conserved Core of a HumanSIR2Homologue Functions in Yeast Silencing

Sherman

Stone

Freeman-Cook

et al. 1999

MBoC

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Silencing is a universal form of transcriptional regulation in which regions of the genome are reversibly inactivated by changes in chromatin structure. Sir2 (Silent Information Regulator) protein is unique among the silencing factors in Saccharomyces cerevisiae because it silences the rDNA as well as the silent mating-type loci and telomeres. Discovery of a gene family of Homologues of Sir Two (HSTs) in organisms from bacteria to humans suggests that SIR2's silencing mechanism might be conserved. The Sir2 and Hst proteins share a core domain, which includes two diagnostic sequence motifs of unknown function as well as four cysteines of a putative zinc finger. We demonstrate by mutational analyses that the conserved core and each of its motifs are essential for Sir2p silencing. Chimeras between Sir2p and a human Sir2 homologue (hSir2Ap) indicate that this human protein's core can substitute for that of Sir2p, implicating the core as a silencing domain. Immunofluorescence studies reveal partially disrupted localization, accounting for the yeast-human chimeras' ability to function at only a subset of Sir2p's target loci. Together, these results support a model for the involvement of distinct Sir2p-containing complexes in HM/telomeric and rDNA silencing and that HST family members, including the widely expressed hSir2A, may perform evolutionarily conserved functions.

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Antibodies specific to acetylated histones document the existence of deposition- and transcription-related histone acetylation in Tetrahymena.

Cited by 139 publications

References 30 publications

Pepsin-Mediated Processing of the Cytoplasmic Histone H2A to Strong Antimicrobial Peptide Buforin I

Pepsin-Mediated Processing of the Cytoplasmic Histone H2A to Strong Antimicrobial Peptide Buforin I

Transcriptionally Active Xenopus laevis Somatic 5 S Ribosomal RNA Genes Are Packaged with Hyperacetylated Histone H4, Whereas Transcriptionally Silent Oocyte Genes Are Not

The Conserved Core of a HumanSIR2Homologue Functions in Yeast Silencing

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