Biochemical and immunological characterization of two distinct variants of histone H2A in Friend leukemia

Blankstein, Larry; Stollar, B D; Franklin, Samuel G.; Zweidler, Alfred; Levy, Stuart B.

doi:10.1021/bi00640a003

Cited by 52 publications

(20 citation statements)

References 29 publications

(28 reference statements)

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“…The change Leu-Met at position 54 has already been found in cuttlefish histone H2A [ 6 ] ; but more frequently this change occurs at position 51 as observed in H2A from mouse ascite tumor cells [25], mouse Friend leukemia cells [26] and sea-urchin embryos [27].…”

Section: Discussionmentioning

confidence: 86%

Primary Structure of Histone H2A from Gonads of the Starfish Asterias rubens

Martinage

Bélaïche

Dupressoir

et al. 1983

European Journal of Biochemistry

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The complete amino acid sequence (124 residues) of histone H2A from gonads of the starfish Asterius ruhens has been established from automated sequence analyses of large fragments obtained by staphylococcal protease digestion of histone H2A and by limited hydrolysis of H2A-H2B complex with mouse submaxillary gland protease and from structural studies of peptides generated by enzymatic hydrolyses of these fragments or of the protein.By comparison with calf homologous histone, the starfish histone H2A shows 5 deletions and 12 substitutions. Half of the substitutions are non-conservative.Microheterogeneities were found at positions 18, 40 and 50 and result in the existence of at least two variants of starfish gonad histone H2A.Structural studies of histones H2A from different species widely separated on the evolutionary scale [I -61 have shown that extensive structural modifications (point mutations, deletions and insertions) mainly occur in the highly basic N-terminal and C-terminal parts of the protein. The central region of the molecule (residues 18-118), which strongly interacts with the region 32-125 of histone H2B to form the dimer H2A-H2B in the nucleosome, appears generally conserved during evolution.The higher degree of variability of histone H2B in Echinidae [7], one of the three families of the echinoderms has led us to investigate the primary structure of both histones H2A and H2B in Asterinidae.In this paper we report the complete amino acid sequence of histone H2A from gonads of the starfish Asterias ruhens.

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Section: Discussionmentioning

confidence: 86%

Primary Structure of Histone H2A from Gonads of the Starfish Asterias rubens

Martinage

Bélaïche

Dupressoir

et al. 1983

European Journal of Biochemistry

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“…Nonetheless, a number of histone variants are known (2)(3)(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.…”

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confidence: 99%

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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“…MEL cells contain at least five H2a variants with a variety of primary sequence differences (5,6,20). The bulk of the H2a population is made up of two species: H2a.1, a replication-dependent variant which decreases from 47 to about 23% of all H2a's after 4 days of differentiation, and H2a.2, a partially replication-dependent variant which increases in relative abundance from 48 to 58% (20).…”

Section: Methodsmentioning

confidence: 99%

“…In mammals, each histone subtype, with the possible exception of H4, is represented by several nonallelic primary sequence variants which display distinct patterns of expression during differentiation (5,6,14,20,24,(49)(50)(51)(52)(53)(54). Nuclease digestion studies indicate that these variants are nonrandomly distributed between actively transcribed and inactive DNA sequences, suggesting that the histones may play a role in establishing or at least maintaining different chromatin conformations (35,47,48).…”

mentioning

confidence: 99%

Changes in the levels of three different classes of histone mRNA during murine erythroleukemia cell differentiation.

Brown¹,

Wellman²,

Sittman³

1985

Mol. Cell. Biol.

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We used a gene-specific Si nuclease assay to study the changes in steady-state mRNA levels of several core histone variants during the differentiation of murine erythroleukemia cells. These studies allowed us to distinguish three distinct expression classes of histone genes. The expression of the major replication-dependent class of histone genes was tightly linked to DNA synthesis. The concentrations of these transcripts decreased rapidly as cell division slowed during the process of differentiation. In contrast, the replication-independent H3.3 transcript levels were constitutively maintained throughout differentiation and were unaffected by inhibitors of DNA or protein synthesis. We also identified among the cloned histone genes used as probes a third expression class, the partially replication-dependent variants. Expression of these transcripts became transiently uncoupled from the reduced rate of DNA synthesis accompanying the early stages of differentiation. We show that their synthesis is sensitive to the DNA synthesis inhibitor hydroxyurea but that selective uncoupling from DNA synthesis of these histone mRNAs occurs at a specific stage of differentiation. We present several hypotheses to explain how this might be accomplished. The expression characteristics of the mRNAs studied coincided with those of the proteins for which they code, indicating that changes in the relative levels of the different variants is mediated at least in part by changes in mRNA levels.Histones are a complex family of proteins responsible for organizing the nucleosomal structure of eucaryotic chromatin (22,27,32). In mammals, each histone subtype, with the possible exception of H4, is represented by several nonallelic primary sequence variants which display distinct patterns of expression during differentiation (5,6,14,20,24,(49)(50)(51)(52)(53)(54). Nuclease digestion studies indicate that these variants are nonrandomly distributed between actively transcribed and inactive DNA sequences, suggesting that the histones may play a role in establishing or at least maintaining different chromatin conformations (35,47,48).Histone variants are classified according to their expression characteristics (53, 54). The major class, the replicationdependent variants, predominate in rapidly dividing tissues, and their synthesis is tightly linked to DNA synthesis (7,9,16,34,49). The mRNAs for these variants begin to accumulate near the G1-S border of the cell cycle, reach maximum levels at mid-S phase, and then rapidly decline as DNA synthesis is completed (1,2,8,11,21,33). Regulation of replication-dependent histone mRNAs occurs at the level of transcription and stability (21, 40).Considerably less is known about the regulation of expression of the other classes of histones. In studies of regenerating liver, partially replication-dependent histones have been identified as a group of variants which are induced with the onset of DNA synthesis but not completely repressed after cessation of DNA synthesis (53, 54). The expression of another class ...

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Biochemical and immunological characterization of two distinct variants of histone H2A in Friend leukemia

Cited by 52 publications

References 29 publications

Primary Structure of Histone H2A from Gonads of the Starfish Asterias rubens

Primary Structure of Histone H2A from Gonads of the Starfish Asterias rubens

Drosophila nucleosomes contain an unusual histone-like protein.

Changes in the levels of three different classes of histone mRNA during murine erythroleukemia cell differentiation.

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