The amino acid sequence of wheat histone H2B<sub>(2)</sub>

Wf, Brandt; J, de Andrade Rodrigues; C, von Holt

doi:10.1111/j.1432-1033.1988.tb14033.x

Cited by 20 publications

(12 citation statements)

References 19 publications

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“…3). Applied to wheat and alfalfa H2B variants, this procedure confirmed their assignments to the H2B class, and also confirmed that differences in sizes of H2B variants were due to differences in the sizes of their N-terminal regions, as previously reported by Brandt et al (5).…”

Section: P_supporting

confidence: 67%

“…Alfalfa H2A amino acid sequences have not been determined, but the electrophoretic behavior and peptide maps of alfalfa H2A(,2) closely resemble those of wheat H2A(,2), indicating that their amino acid sequences are similar.Phosphorylation sites were localized by autoradiography of the NBS peptides (32P; Fig. 2 (5), vary in mol wt; they are designated H2B()-H2B (6) in this report ( Fig. la; the subscripts may not correspond to those previously assigned due to differences in gel electrophoresis methods).…”

Section: P_mentioning

confidence: 99%

“…A hypothesis accounting for the presence of large H2A and H2B histone variants in plants and phosphorylation of plant H2A C-terminal regions is proposed. The utility of S-P tetrapeptides for modulation of DNA-protein interactions is discussed.H2A and H2B histone variants from wheat contain long N-and C-terminal regions, rich in basic amino acids, hydroxyamino acids, proline and alanine (5,17,18). These regions resemble the DNA-binding 'head' and 'tail' regions of HI histones (1).…”

mentioning

confidence: 99%

“…H2A and H2B histone variants from wheat contain long N-and C-terminal regions, rich in basic amino acids, hydroxyamino acids, proline and alanine (5,17,18). These regions resemble the DNA-binding 'head' and 'tail' regions of HI histones (1).…”

mentioning

confidence: 99%

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Phosphorylation of Plant H2A Histones

Green¹,

Gustavsen²,

Poccia³

1990

Plant Physiol.

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Phosphorylation of wheat (Triticum aestivum) and alfalfa (Medicago sativa) H2A histone variants was examined during early seedling growth. The C-terminal regions of wheat H2A variants contain multiple S-P tetrapeptides (senne-proline adjacent to a pair of basic amino acids) which resemble known phosphorylation sites in histones from other species. Phosphorylation of nucleosomal core histones was assessed by autoradiography of proteins labeled in vivo with 32pi and resolved by two-dimensional polyacrylamide gel electrophoresis, and phosphorylation sites were mapped by cleaving in vivo labeled H2A variants with Nbromosuccinimide. Essentially all phosphorylation of nucleosomal core histones in wheat and alfalfa seedlings occurred within the C-terminal peptides obtained from wheat and alfalfa H2A variants. A hypothesis accounting for the presence of large H2A and H2B histone variants in plants and phosphorylation of plant H2A C-terminal regions is proposed. The utility of S-P tetrapeptides for modulation of DNA-protein interactions is discussed.H2A and H2B histone variants from wheat contain long N-and C-terminal regions, rich in basic amino acids, hydroxyamino acids, proline and alanine (5,17,18). These regions resemble the DNA-binding 'head' and 'tail' regions of HI histones (1). Located periodically in the C-terminal regions of wheat H2A variants are two or three S-P tetrapeptides (serineproline adjacent to a pair of basic amino acids) (17, 18). Similar S-P tetrapeptides are known phosphorylation sites in the C-terminal regions of vertebrate HI histones (reviewed in Poccia [ 13]) and in the long N-terminal regions of sea urchin sperm HI and H2B histone variants, Sp HI and Sp H2B (7, 16).These observations prompted a study of nucleosomal core histone phosphorylation in early seedlings, during a period of rapid cell growth which follows a protracted period of nuclear quiescence. The results show that nearly all nucleosomal histone phosphorylation occurs within C-terminal peptides obtained from H2A histone variants, suggesting that S-P tetrapeptides are the primary sites ofphosphorylation, as they are in the sea urchin sperm variants Sp HI and Sp H2B. By analogy with Sp H1 and Sp H2B, we propose a hypothesis which accounts for the presence of large H2A and H2B histone variants in plants and C-terminal phosphorylation of the H2A variants. MATERIALS AND METHODSGermination, Seedling Growth, and Radiolabeling Seeds ofalfalfa (Medicago sativa) and hard red spring wheat (Triticum aestivum) were purchased from local grocery stores and germinated in Petri dishes on filter paper wetted with distilled water. After 48 h at 22°C, germinated seedlings (2 g, 2-4 mm in length) were transferred to fresh filter paper and wetted with 0.1 mCi/mL 32Pi (orthophosphate in carrier free aqueous solution; Amersham). Seedlings were incubated for an additional 24 h, after which endosperm or cotyledon tissues were removed. The seedlings were washed with cold water to remove residual 32Pi. Subsequent steps were at OC. Histone IsolationW...

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Section: P_supporting

confidence: 67%

Section: P_mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Phosphorylation of Plant H2A Histones

Green¹,

Gustavsen²,

Poccia³

1990

Plant Physiol.

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“…The H2B variants (Fig. 1 b) are the subject of a separate communication [24]. Table 1 shows the amino acid composition of the isohistone H2A(,, type.…”

Section: Resultsmentioning

confidence: 99%

The primary structure of the histone H2A₍₂₎ type from wheat germ

Rodrigues

Brandt

Holt

1988

European Journal of Biochemistry

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The histone H2A,,, type from wheat germ comprises at least two highly homologous isohistones with 151 amino acid residues. Microheterogeneity occurs mainly at the N-terminal and C-terminal regions, These isohistones have both N-terminal(7 amino acid residues) and C-terminal(l5 amino acid residues) extensions relative to calf thymus histone H2A.Histones are essential for the ordered packing of DNA into nucleosomes and higher-order structures [I]. The widespread occurrence of isohistones (for review see [2, 31) suggests that nucleosomes are heterogeneous with respect to their histone content. In animal systems, different sets of the non-allelic histone variants are expressed coordinately at different stages of development and cell differentiation [3, 41. Less is known about the structural variation of plant histones, and there is no evidence yet on the existence or otherwise of a developmental program , and the other, H2AC2,, with blocked N-termini exhibiting both N-terminal and C-terminal extensions. Each type comprises at least two highly homologous species which only becomes apparent on sequencing. Neither HPLC, ion-exchange chromatography nor Triton X-IOO/PAGE has succeeded in separating the closely related iso-histones within each type from each other. We report here the characterization of the structure of the H2A,,, type isohistone from wheat germ. MATERIALS AND METHODS Isolation of histone H2A,,,Wheat germ (Triticum aestivum) was obtained from Sasko Mills (Rondebosch, Cape). This material contained at least 80% of the cultivar T4. Chromatin was isolated essentially according to the method of Johns and Butler [9] with some modifications as described previously [6].Exclusion chromatography of histones was performed at room temperature on a column (4.5 x 80 cm) of Bio-Gel P-60 (100-200 mesh) as described previously buffer (pH 5.30) containing 6 M urea. A linear NaCl gradient was used to elute the fractions (Fig. 1 a). After desalting, the H2A[,,-containing fraction was further purified on a Bio-Gel P-60 column equilibrated with 20 mM HC1, 50 mM NaCl as described previously [7]. The eluant contained in addition, 10 mM sodium bisulphite, 1 mM phenylmethylsulfonylfluoride and 1 mM N-a-tosyl-L-lysylchloromethane, solubilized first in a small volume of propan-1-01. Analytical proceduresPAGE was performed either in acid/urea/Triton X-100 gels according to Zweidler [9] or in SDS according to Laemmli [lo]. Amino acid compositions were determined after 24 h hydrolysis in 5.7 M HCl containing 0.025% (massivol.) phenol. No correction factors for incomplete hydrolysis or destruction of amino acids were applied.Cleavage of the acetylated N-terminal methionine residue with CNBr [ll] was performed as described previously [6]. Peptides were generated through enzymatic cleavage employing trypsin (after prior citraconylation of the isohistone), endoproteinase Lys-C, and Staphylococcus aureus V8 protease as described previously for isohistone H2A,,, [7]. The peptides generated through ezymatic cleavages were fractionated by g...

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