Cell-cycle-dependent dissociation of histone H1 from chromatin in nuclei of P. polycephalum

Cruickshank, Helen M.; Walker, I.O.

doi:10.1093/nar/9.15.3873

Cited by 5 publications

(4 citation statements)

References 28 publications

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“…Mohberg & Rusch later (1970) published an amino acid composition for Physarum band 1 that was essentially correct although marred by the presence of an "unidentified acidic amino acid" that can now be seen to be due to oxidation during the hydrolysis (Corbett, 1979). Similar amino acid analyses of Physarum histone HI from plasmodia have been published by Tyrsin et al (1977a), Fischer & Laemmli (1980), and Cruikshank & Walker (1981). A comparison of the Physarum histone H1 amino acid analysis published here (Table III) with an amino acid analysis of calf thymus HI (Johns, 1976) clearly shows the similarity, but there are also substantial differences.…”

Section: Discussionsupporting

confidence: 81%

“…Band 4 was shown to contain two components. The amino acid composition of HI was determined and has been confirmed in several laboratories (Matthews & Bradbury, 1982; Fischer & Laemmli, 1980; Cruikshank & Walker, 1981; Cote et al, 1982). While we have found the CaCl2 extraction method very useful for studies of histones HI and H4, it has been criticized by others (Fischer & Laemmli, 1980;Gurley et al, 1978), and we have found variable yields of histone H3.…”

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confidence: 80%

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Isolation, identification, and characterization of histones from plasmodia of the true slime mold Physarum polycephalum using extraction with guanidine hydrochloride

Mende¹,

Waterborg²,

Mueller³

et al. 1983

Biochemistry

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Histones from plasmodia of the true slime mold Physarum polycephalum have been prepared free of slime by an approach to histone isolation that uses extraction of nuclei with 40% guanidine hydrochloride and chromatography of the extract on Bio-Rex 70. This procedure followed by chromatography or electrophoresis has been used to obtain pure fractions of histones from Physarum microplasmodia. Physarum microplasmodia have five major histone fractions, and we show by amino acid analysis, apparent molecular weight on three gel systems containing sodium dodecyl sulfate, mobility on gels containing Triton X-100, and other characterizations that these fractions are analogous to mammalian histones H1, H2A, H2B, H3, and H4. Significant differences between Physarum and mammalian histones are noted, with histone H1 showing by far the greatest variation. Histones H1 and H4 from Physarum microplasmodia have similar, but not identical, products of partial chymotryptic digestion compared with those of calf thymus histones H1 and H4. Labeling experiments, in vivo, showed that histone H1 is the major phosphorylated histone and approximately 15 separate phosphopeptides are present in a tryptic digest of Physarum histone H1. The core histones from Physarum, histones H2A, H2B, H3, and H4, are rapidly acetylated; histone H4 shows five subfractions, analogous to the five subfractions of mammalian histone H4 (containing zero to four acetyllysine residues per molecule); histone H3 has a more complex pattern that we interpret as zero to four acetyllysine residues on each of two sequence variants of histone H3; histones H2A and H2B show less heterogeneity. Overall, the data show that Physarum microplasmodia have a set of histones that is closely analogous to mammalian histones.

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Section: Discussionsupporting

confidence: 81%

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confidence: 80%

Isolation, identification, and characterization of histones from plasmodia of the true slime mold Physarum polycephalum using extraction with guanidine hydrochloride

Mende¹,

Waterborg²,

Mueller³

et al. 1983

Biochemistry

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“…Following digestion of D. discoideum nuclei and nucleoli with micrococcal nuclease, two populations of mononucleosomes are produced (23,(25)(26)(27)(28). One population containing 146 bp DNA corresponds to the core particle found in all eukaryotes.…”

Section: _mentioning

confidence: 99%

“…2% TCA) and in having 20.5% acidic amino acids. Lysine-rich, major nucleoproteins containing between 14 and 19% glutamic and aspartic acids have been described in Physarum polyceiphalum (23)(24)(25)(26). Similar proteins isolated from Saccharomyces cerevisae have been called HMG proteins (27,28) or Hi (29)(30)(31)(32)(33)(34)(35), although their role in chromatin is unknown.…”

Section: Introductionmentioning

confidence: 99%

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

Parish

Schmidlin

1985

Nucl Acids Res

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Mononucleosomes released from Dictyostelium discoideum chromatin by micrococcal nuclease contained two distinctive DNA sizes (166-180 and 146 bp). Two dimensional gel electrophoresis suggested a lysine-rich protein protected the larger mononucleosomes from nuclease digestion. This was confirmed by stripping the protein from chromatin with Dowex resin. Subsequently, only the 146 bp mononucleosome was produced by nuclease digestion. Reconstitution of the stripped chromatin with the purified lysine-rich protein resulted in the reappearance of the larger mononucleosomes. Two-dimensional gel electrophoresis showed the protein was associated with mononucleosomes. Hence, the protein functions as an H1 histone in bringing the two DNA strands together at their exit point from the nucleosome. Trypsin digestion of the lysine-rich protein in nuclei resulted in a limiting peptide of approx. 10 kilodaltons. Trypsin concentrations which degraded the protein to peptides of 12-14 kilodaltons and partially degraded the core histones did not change the DNA digestion patterns obtained with micrococcal nuclease. Thus, the trypsin-resistant domain of the lysine-rich protein is able to maintain chromatosome structure.

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The Core Histones from the Slime Mold Physarum polycephalum

Champagne

Demouveau

Couppez³

et al. 1982

European Journal of Biochemistry

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The histones of Physarum polycephalum have been isolated and fractionated using a combination of gel exclusion and ion-exchange chromatography or differential precipitation. The four core histones were unambiguously identified by gel electrophoresis, amino acid composition and N-terminal analysis.The molecular weight of Physarum histones H3, H2B and H4 are very close or identical to the corresponding histones from chicken erythrocytes. Physarum H2A is significantly larger than chicken erythrocyte H2A and its N-terminal residue is alanine. From the differences in amino acid compositions and in the peptide maps between Physarum and calf H4, one can expected some changes in the amino acid sequence of Physarum H4.The slime mold Physarum polycephalum is widely used as a model system for cell cycle studies [I]. In most instances, information gained in studying Physarum can be extrapolated to more complex eukaryotic organisms. However, to our knowledge, the four 'core histones' of Physarum have never been described unambiguously. As pointed out by Mohberg and Rusch [15], Physarum histones are difficult to isolate. The nuclei are extremely resistant to breakage and frequently contain an acidic polysaccharide which forms complexes with the histones.Physarum histones were named by Mohberg and Rusch [15] in reverse order of their electrophoretic mobilities in an acidlurea system: P 1, 2a, 2b, 3, 4, 5 and 6 [16-201. The molecular weights of these fractions were determined by electrophoresis on polyacrylamide gels containing sodium dodecyl sulfate [17,18]. Gel filtration chromatography was used with limited success and, on the basis of amino acid analysis and peptide mapping [18,19], the principal histone fractions P1, P4a, P4b, P5 and P6 were assumed to be homologous to the histones H1, H3, H2B, H2A and H4 of calf thymus, respectively. More recently it was suggested that histone P3 of Physarum and histone H2B from calf thymus have regions of homology [20].We describe here a method for purifying the four core histones of Physarum polycephalum. These proteins were characterized by polyacrylamide gel electrophoresis, amino acid analysis, end-group determination and peptide mapping. We compare the data to those obtained with other known histones. Finally we conclude that histone P3 from Physarum polycephalum is homologous to histone H2A. MATERIALS AND METHODSCulture of Physarum polycephalum Plasmodia as described by Daniel and Rusch [21].Macroplasmodia were grown in petri dishes for two days Nuclei were suspended in 1 M CaCI2 (60 ml for ten macroplasmodia) and 1 mM phenylmethylsulfonyl fluoride was added. Nuclei were lysed for 5 min at 80°C, cooled in ice and centrifuged at 4 "C for 30 min at 40000 x g. HCI was added to the supernatant to a final concentration of 0.25 M.The mixture was stirred at 4°C for 1 h, then centrifuged 30 min at 40000 x g.Histones were precipitated from the supernatant by 25 trichloroacetic acid at 4 "C overnight, centrifuged at 40000 x g, washed with acidified acetone, then with acetone, and finally dri...

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Cell-cycle-dependent dissociation of histone H1 from chromatin in nuclei of P. polycephalum

Cited by 5 publications

References 28 publications

Isolation, identification, and characterization of histones from plasmodia of the true slime mold Physarum polycephalum using extraction with guanidine hydrochloride

Isolation, identification, and characterization of histones from plasmodia of the true slime mold Physarum polycephalum using extraction with guanidine hydrochloride

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

The Core Histones from the Slime Mold Physarum polycephalum

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