Biochemical evidence for a distinctive chromatin structure in nucleoli of <i>dictyostelium</i>

Widmer, Rosa M.; Fuhrer, Stelia; Parish, Roger W.

doi:10.1016/0014-5793(79)80533-5

Cited by 16 publications

(6 citation statements)

References 36 publications

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“…Protein was recovered from the phenol phase and interphase by adding HC1 to 1N and 10 vol acetone. DNA and proteins were separated on gels as previously described (35,36).…”

Section: Siaterials and Methodsmentioning

confidence: 99%

“…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. Neurospora crasse Hi contains 15.7% acidic amino acids (32).…”

Section: Introductionmentioning

confidence: 99%

“…The DNA of D.disg ideum is organised into nucleosomes (33)(34)(35)(36)(37)(38)(39)(40) and we wished to discover whether the lysine-rich protein was involved in the structure of the 100 A fiber. We had observed that mononucleosome populations obtained by micrococcal nuclease digestion contained two DNA sizes, one equivalent to the core (146 bp) and the second somewhat larger (166-180 bp) (33, [35][36][37][38]. One interpretation was that the larger sized particles carried a protein resembling Hi in function.…”

Section: Introductionmentioning

confidence: 99%

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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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“…Protein was recovered from the phenol phase and interphase by adding HC1 to 1N and 10 vol acetone. DNA and proteins were separated on gels as previously described (35,36).…”

Section: Siaterials and Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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

Parish

Schmidlin

1985

Nucl Acids Res

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“…Micrococcal nuclease digestion was performed by resuspending washed nuclei (1.5 x 10 /ml) from vegetative or differentiating cells in 3 ml of 20 mM4 Tris-HCl, pH 7.8, 0.5 mM CaCl2 and incubating at 250C with 12.5 units/ml enzyme (37). Incubation times varied from 15 sec to 5 min.…”

Section: Micrococcal Nuclease Digestionmentioning

confidence: 99%

Hypersensitive sites in the 5′ and 3′ flanking regions of the cysteine proteinase I gene ofDictyostelium discoideum

1986

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“…Washed nuclei (1.5 x 109 per ml) (25) were resuspended in 3 ml 20 mM Tris-HCl, pH 7.8, 0.5 mM CaCl2 and incubated at 250C with either 80 units/ml (nuclei from vegetative cells) or 50 units/ml (differentiating cells) micrococcal nuclease (26). Incubation times varied from 15 s to 5 min.…”

Section: Micrococcal Nuclease Digestionmentioning

confidence: 99%

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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Biochemical evidence for a distinctive chromatin structure in nucleoli of dictyostelium

Cited by 16 publications

References 36 publications

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

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

Hypersensitive sites in the 5′ and 3′ flanking regions of the cysteine proteinase I gene ofDictyostelium discoideum

The effects of transcription on the nucleosome structure of fourDictyosteliumgenes

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