A lysine-rich protein functions as an H1 histone in<i>Dictyostelium discoideum</i>chromatin

Parish, Roger W.; Schmidlin, Sylvia

doi:10.1093/nar/13.1.15

Cited by 5 publications

(3 citation statements)

References 59 publications

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“…An optimized acid-extraction and electrophoresis protocol for histones from vegetatively growing Dictyostelium cells ( 27 ) generated preparations enriched in five major coomassie-stainable protein bands in the region where histones normally migrate on SDS–polyacrylamide gels ( Figure 1 A). These bands were subject to mass spectrometry ( Figure 1 B) and we identified histone H2AX [Band 2 ( 27 )], H2Bv3 (Band 1) and H4 (Band 5), as previously reported ( 17 ) and also histone H1 [Band X; ( 30 , 31 )]. The two genes encoding H4 encode proteins of identical amino acid sequence, so we have not been able to determine if both are expressed by this analysis.…”

Section: Resultssupporting

confidence: 70%

“…These bands were subject to mass spectrometry (Figure 1B) and we identified histone H2AX [Band 2 (27)], H2Bv3 (Band 1) and H4 (Band 5), as previously reported (17) and also histone H1 [Band X; (30,31)]. The two genes encoding H4 encode proteins of identical amino acid sequence, so we have not been able to determine if both are expressed by this analysis.…”

Section: Resultsmentioning

confidence: 70%

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Dynamic acetylation of lysine-4-trimethylated histone H3 and H3 variant biology in a simple multicellular eukaryote

Hsu

Chubb

Muramoto

et al. 2012

Nucleic Acids Research

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Dynamic acetylation of all lysine-4-trimethylated histone H3 is a complex phenomenon involved in Immediate-early gene induction in metazoan eukaryotes. Higher eukaryotes express repeated copies of three closely related H3 variants, inaccessible to genetic analysis. We demonstrate conservation of these phenomena in Dictyostelium which has three single-copy H3 variant genes. Although dynamic acetylation is targeted to two H3 variants which are K4-trimethylated, K9-acetylation is preferentially targeted to one. In cells lacking Set1 methyltransferase and any detectable K4-trimethylation, dynamic acetylation is lost demonstrating a direct link between the two. Gene replacement to express mutated H3 variants reveals a novel interaction between K4-trimethylation on different variants. Cells expressing only one variant show defects in growth, and in induction of a UV-inducible gene, demonstrating the functional importance of variant expression. These studies confirm that dynamic acetylation targeted to H3K4me3 arose early in evolution and reveal a very high level of specificity of histone variant utilization in a simple multicellular eukaryote.

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Section: Resultssupporting

confidence: 70%

Section: Resultsmentioning

confidence: 70%

Dynamic acetylation of lysine-4-trimethylated histone H3 and H3 variant biology in a simple multicellular eukaryote

Hsu

Chubb

Muramoto

et al. 2012

Nucleic Acids Research

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“…A series of bands was generated with a repeat of around 180 bp. The mononucleosomes gave two DNA bands, the core size DNA (1146 bp) and the chromatosome plus linker DNA (166-180 bp) which carries an Hi-like protein (37). The band pattern did not change during differentiation.…”

Section: Micrococcal Nuclease Digestionmentioning

confidence: 95%

The effects of transcription on the nucleosome structure of fourDictyosteliumgenes

Pavlovic¹,

Banz²,

Parish³

1989

Nucl Acids Res

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Micrococcal nuclease digestion of Dictyostelium discoideum nuclei from various developmental stages was used to investigate transcription-related changes in the chromatin structure of the coding region of four genes. Gene activity was determined by Northern blotting and nuclear run on experiments. During strong transcription of the developmentally regulated cysteine proteinase I gene, a smear superimposed on a nucleosomal ladder was observed, indicating perturbation of nucleosomal structure was occurring. However, two other developmentally regulated genes, discoidin I and pSC253, showed only slight nucleosome disruption during high levels of transcription. The chromatin structure of a fourth gene (pCZ22) was disrupted throughout development, even at those stages where transcription was greatly reduced. We suggest that although nucleosome structure can be transiently perturbed by the passage of the transcription complex in vivo, the degree of perturbation and the speed with which nucleosomes reassemble is also influenced by the DNA sequence.

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The histones ofDictyostelium discoideum

Garside

Maclean

1987

Experientia

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Summary. The histones of the cellular slime mould Dictyostelium discoideum have been separated by electrophoresis using both acid urea and sodium dodecyl sulphate systems, and the gel pattern compared with that of histones from Physarum polycephalum and calf thymus. Dictyostelium is found to possess a full complement of H1, H2A, H2B, H3 and H4.Key words. Dictyostelium; slime mould; histone; electrophoresis.Dictyostelium discoideum, the cellular slime mould, is an extensively studied organism. Its easy growth and culture, the existence of axenic strains of free living amoebae, such as AX-2, which may be cultured indefinitely in liquid suspension cultures, and its small genome size, combine to make the organism interesting and important in the context of biochemistry and genetics. Work on the genome of Dictyostelium has focused attention on its ehromatin, and many laboratories have attempted separation and characterization of its histones ~4.Studies on Dictyostelium histones are of interest since this organism is a primitive eukaryote and there has been some doubt about whether such organisms have the full complement of histones as found in higher eukaryotes. The paths of histone evolution are also interesting, especially since core histone H3 and H4 are the most highly conserved of all proteins whilst H1 is less strongly conserved and, at least in higher organism, is coded for by a small multi gene family. Since the genome of this organism is small, the yield of histones is also small, and identification of histones by comprehensive comparison of the amino acid composition with that of other lower eukaryotes such as Physarum, has not as yet proved possible. We have, therefore, sought to achieve good separation of Dietyostelium histones with both triton/acetic acid urea gels and sodium dodecyl sulphate gels, and also to use a two-dimensional gel electrophoresis separation, somewhat similar to that advocated by Bonner et a12 and thus to proceed to a more accurate identification of these histones than has been previously obtained. In addition, our histones have been extracted from nuclei isolated by the use of digitonin, a natural plant saponin, rather than with nonidet P40. We have already shown that, using a number of criteria, such nuclei are superior to those isolated with NP-40 ~~ and, in particular, that their DNA is not substantially degraded. Materials and methodsGrowth conditions. Dictyostelium discoideum strain AX-2 cells were grown in suspension culture as previously described ~~ and nuclei were isolated from such cells using the 0.1 mg/ml digitonin method described in the same paper.Isolation ofhistones. Basic nucleoproteins were extracted from isolated nuclei by one of two methods. 1) The acid extraction of Bakke and Bonner 4. 2) The method of Mohberg and Rusch using the modification of Coukell and Walker H the suspension of nuclei being heated in 1 M CaClz at 80~ for 15 min. In both methods basic nucleoproteins were precipitated by adding 4 vol. of ethanol at -20 ~ and allowing precipitation to proceed ...

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A lysine-rich protein functions as an H1 histone inDictyostelium discoideumchromatin

Cited by 5 publications

References 59 publications

Dynamic acetylation of lysine-4-trimethylated histone H3 and H3 variant biology in a simple multicellular eukaryote

Dynamic acetylation of lysine-4-trimethylated histone H3 and H3 variant biology in a simple multicellular eukaryote

The effects of transcription on the nucleosome structure of fourDictyosteliumgenes

The histones ofDictyostelium discoideum

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