Isolation and characterization of the histone variants in chicken erythrocytes

Urban, Michael K.; Franklin, Samuel G.; Zweidler, Alfred

doi:10.1021/bi00585a017

Cited by 102 publications

(48 citation statements)

References 34 publications

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“…It can, however, be distinguished from both H2A of the microheterogeneity in D2 revealed by acid/urea/Triton DF-16/polyacrylamide gel electrophoresis is unknown. It may be the result of posttranslational modification, or, alternatively, sequence heterogeneity similar to that observed in mammalian or chicken histones (2,32).…”

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confidence: 98%

“…The similarities of size and amino acid composition between D2 and the slightly lysine-rich histones suggest that D2 might substitute for either H2A or H2B. In this respect, D2 may be analogous to the vertebrate proteins Ml and M2 (5,32). It has been suggested that these histone-like proteins substitute for H2A (14,32 The Drosophila histone genes are encoded in a tandem array that is located at 2:39D-E on the polytene salivary gland map (33).…”

mentioning

confidence: 99%

“…In this respect, D2 may be analogous to the vertebrate proteins Ml and M2 (5,32). It has been suggested that these histone-like proteins substitute for H2A (14,32 The Drosophila histone genes are encoded in a tandem array that is located at 2:39D-E on the polytene salivary gland map (33). It will be interesting to learn whether D2 is encoded in a subset of the histone gene cluster or is encoded separately.…”

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confidence: 99%

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Drosophila nucleosomes contain an unusual histone-like protein.

Palmer¹,

Snyder²,

Blumenfeld³

1980

Proc. Natl. Acad. Sci. U.S.A.

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Histones are major structural components of the basic repeating subunit of chromatin, the nucleosome (1). Core histone sequences are remarkably conserved during evolution. Nonetheless, a number of histone variants are known (2-4). Synthesis of core histone variants is stage-specific during sea urchin development (3, 4), tissue-specific in mammals (5), and correlated with erythroid differentiation in Friend leukemia cells (6, 7). These observations suggest that nucleosome heterogeneity is related to the functional specialization of specific elements of chromatin. In this context, it may be important that mononucleosomes isolated from Drosophila chromatin contain large amounts of a histone-like protein that is not a standard core histone. This protein was first identified by Alfageme et al. (8), and designated D2, or "Drosophila 2."In this communication, we demonstrate that D2 is nucleosomal and histone-like, yet is readily distinguished from each of the four "standard" core histones. It is also conserved during the evolution of Drosophila. The curious features of D2 raise questions about histone evolution and the organization of histone genes in Drosophila.MATERIALS AND METHODS Purification of Nuclei. D. melanogaster embryo nuclei and adult head nuclei were prepared with three modifications of published procedures (9, 10): (i) buffer A of Hewish and Burgoyne (11), containing 1 mM EDTA, 0.2 mM ethylene glycol bis(f3-aminoethyl ether)-N,N,N'N'-tetraacetic acid (EGTA), and 0.2 mM phenylmethylsulfonyl fluoride was used for the nuclear isolation; (ii) the crude nuclear pellet was washed in buffer containing 0.1% Triton X-100; (iii) the detergent-washed nuclei were then resuspended and recentrifuged three times in buffer lacking Triton X-100.Purification of Mononucleosomes. Nuclei were suspended in buffer A of Hewish and Burgoyne (11), made 4 mM in CaCl2, and digested at 250C to 25-29% acid solubility with staphylococcal nuclease (EC 3.1.31.1). Digested nuclei were lysed according to Noll and Kornberg (12). Digests were fractionated by sedimentation through linear (5-20%) sucrose gradients containing 0.5 M NaCl (13) or by electrophoresis in 5% acrylamide gels (14).Analysis of Nucleosomal DNA. DNA was extracted from pooled, sucrose-gradient-purified nucleosomes (15), omitting RNase treatment. DNA was released from electrophoretically purified nucleosomes with 1% NaDodSO4 and proteinase K at 100 ,ug/ml. DNA was electrophoresed on 5% acrylamide slab gels (16) and visualized by staining with ethidium bromide.Analysis of Nucleosomal Proteins. Gradient-purified nucleosomes were precipitated with 10 mM MgCl2 (17) and extracted with protamine/urea/acetic acid (18). The released chromosomal proteins were applied directly to acetic acid/ urea/Triton DF-16 gels (see below). Electrophoretically purified nucleosomes were electrophoresed into dialysis bags and extracted with 0.25 M HCO (8). The extracted proteins were analyzed by two-dimensional electrophoresis (see below). Purification of Histones. D. melanogaster embryo histones w...

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Drosophila nucleosomes contain an unusual histone-like protein.

Palmer¹,

Snyder²,

Blumenfeld³

1980

Proc. Natl. Acad. Sci. U.S.A.

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“…Expression of this class of genes may be triggered by the onset of DNA replication but is not completely repressed after DNA synthesis. The third class of genes code for replication-independent variants, also termed replacement histones, due to their pattern of expression during development and differentiation (45,52,55,57). In systems of low cellular turnover, these constitutively expressed variants can represent an important proportion of the histone moiety (53).…”

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confidence: 99%

Regulation of histone and beta A-globin gene expression during differentiation of chicken erythroid cells.

Affolter¹,

Côté²,

Renaud³

et al. 1987

Mol. Cell. Biol.

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The expression of the genes for several histones and pA-globin was examined in the chicken erythroid cell lineage. During the transition from CFU-(E) to the mature erythrocyte, histone H5 gradually increased fourfold in nuclei with little concomitant displacement of the Hi histones. This resulted in a 70% net increase in linker histone (Hi plus H5) content. The differential accumulation of H5 reflected (i) an increase in the transcriptional activity of the H5 gene occurring at the erythroblast stage, (ii) an apparent longer half-life of H5 mRNA, and (iii) a higher stability of the protein. Although the transcriptional activity of the histone genes (except H5) decreased with cell age, it was not tightly coupled to the S phase. On the other hand, the mRNA levels for these histones were tightly regulated during the cell cycle. Use of protein and DNA synthesis inhibitors indicated that the content of H5 mRNA was regulated at the posttranscriptional level by a control mechanism(s) differing from those for the other histones. Although the transcription rates of the H5 and pA_globin genes were comparable, differential accumulation of iA_globin mRNA led to a 30-to 170-fold-higher copy number of the pA-globin mRNA as the cell matured.

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“…Sample preparation: Nuclear samples were prepared from whole livers of either rats or mice (13) using a modification of a nuclear isolation technique attributed to Zweidler et al (20). Tissue was quick frozen on dry ice, minced and homogenized.…”

Section: Methodsmentioning

confidence: 99%

A microsample collection device for electrostatically sorted cells or particles and its preparative use for biochemical analysis

Hinson¹,

Pipkin²,

Hudson³

et al. 1982

Cytometry

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Biochemical analysis of proteins extracted from mammalian cell nuclei which were sorted according to DNA content was greatly simplified using a special collection device to recover sorted nuclei. The device was developed for use with a sorting flow cytometer to provide sufficient numbers of particles to achieve adequate sensitivity and resolution while minimizing sample degradation and loss. The design, evaluation and application of the collection device were described with emphasis on sample preparation.Nuclear populations Go + GI, S, and Gz + M, were sorted from regenerating rat liver tissue preparations and collected separately in the special chambers. Nuclear proteins were extracted and analyzed by polyacrylamide gel isoelectric focusing or capillary isotachophoresis.

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Isolation and characterization of the histone variants in chicken erythrocytes

Cited by 102 publications

References 34 publications

Drosophila nucleosomes contain an unusual histone-like protein.

Drosophila nucleosomes contain an unusual histone-like protein.

Regulation of histone and beta A-globin gene expression during differentiation of chicken erythroid cells.

A microsample collection device for electrostatically sorted cells or particles and its preparative use for biochemical analysis

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