The Primary Structure of Histone H1 from Sperm of the Sea Urchin Parechinus angulosus. 2. Sequence of the C-Terminal CNBr Peptide and the Entire Primary Structure

Wn, Strickland; Strickland, Marie; Wf, Brandt; C, von Holt; Lehmann, Arnold; Wittmann‐Liebold, Brigitte

doi:10.1111/j.1432-1033.1980.tb04460.x

Cited by 109 publications

(46 citation statements)

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“…6). The high degree of homology between the C-terminal extension of a core histone and a region in the C-terminal tail of a histone H1 [30] suggests that these two structures may be involved in a similar interaction with DNA, either with linker or core-associated DNA of their own, or even more effectively of neighbouring, nucleosomes to result in a more condensed structure. r I n t a c t H2A,,, C y c l e no.…”

Section: Resultsmentioning

confidence: 99%

The amino acid sequence of wheat histone H2A(1). A core histone with a C-terminal extension

A¹,

Wf²,

C³

1985

Eur J Biochem

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1. The complete amino acid sequence (145 residues) of histone variant H2A,,, from wheat germ Triticum 2. The sequence of histone variant H2A(,, differs from the homologous calf histone in 61 amino acid positions. aestivum cultivar T 4 has been established from Edman degradation of large overlapping fragments.These differences include an extension of H2A,,, by 19 amino acids at its carboxyl end.Histones in general are considered to play a structural rather than a regulatory role in chromatin [l], however the widespread occurrence of histone variants (isohistones), (for review see [2,31) suggests that variants may modulate the conformation of nucleosomes, and in this way modify their susceptibility to regulatory factors. We have established previously the presence of a number of isohistones of the H2A class in wheat embryos [2, 41. We are reporting now the complete structure of one of these isohistones. This structure constitutes a new histone type in that its C-terminal region is extended beyond the size hitherto considered typical for core histones of that group. 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 T 4. Chromatin was isolated essentially according to the method of Johns and Butler [5] with some modifications as described previously [4].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). Approximately 200 mg of total acid-extracted (0.2 M H2S04) histone was dissolved in 10 ml of 6 M urea containing 2% (v/v) 2-mercaptoethanol, loaded onto the column and eluted with 0.02 M HCI (Fig. 1 a).Cation-exchange chromatography was carried out using LKB CM-Trisacryl M equilibrated with 0.05 M sodium acetate adjusted to pH 5.00 with HC1. A linear NaCl gradient was used to elute the H2A(,,-enriched fraction B (Fig. 1 b). Fraction B was then further purified by passage over a BioGel P-60 column equilibrated with 0.02 M HCl, 0.05 M NaCl as described previously [6]. The eluant contained in addition, 10 mM sodium bisulfite, 1 mM PhMeS02F and 1 mM Natosyllysylchloromethane, solubilized first in a small volume of propan-1-01 (Fig. 1 c). The H2A(,,-containing fraction was sometimes recycled over the P-60 column if there was still some contamination evident on gel electrophoresis. Analytical proceduresPolyacrylamide gel electrophoresis (PAGE) employing the detergent Triton X-100 was performed in glass tubes (9 x 0.5 cm) [7, 81.Histone samples were hydrolysed under vacuum for 24 h at 110°C in constant-boiling HCl containing 0.025% (w/v) phenol as antioxidant unless otherwise stated. The analyses were done with either ninhydrin methodology (Beckman 119 automatic analyser) or using hypochlorite and o-phthalaldehyde post-column fluorescence detection (Waters Associates HPLC amino acid analyser [9]).Fragmentation of H2A(,, N-Bromosuccinimide (SucNBr) cleavage [lo] was performed as described previously [ll]. The increase...

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Section: Resultsmentioning

confidence: 99%

The amino acid sequence of wheat histone H2A(1). A core histone with a C-terminal extension

A¹,

Wf²,

C³

1985

Eur J Biochem

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“…Sequcnces used to calculate the number of residucs were chicken FI1 [16], chicken H5 [17], Echinus esculentus sperm I11 [I I] (and C. S . Hill, unpublished; very similar to Purechinus ungulosus sperm H1 [18]). The positive charge density was calculated as the number of lysine and arginine residues divided by the total number of residues.…”

Section: Dna Condensation and Bendability In Histone-dna Complexesmentioning

confidence: 99%

“…40 pg DNA prepared as described above, in 150 pl of a solution containing 18 mM MgCI2, 4.5 mM dithiothreitol, 45 mM KCl, 25 mM imidazole/HCl, pH 6.6, 300 pM ADP, 10 pM ATP, 0.11 mg/ ml autoclaved gelatin, was incubated with approximately 10 pCi [y-32P]ATP (Amersham) and approximately 300 IU T4 polynucleotide kinase at 37°C for 2 h. The radiolabelled DNA was extracted and precipitated with ethanol, washed several times with 70% ethanol, resuspended in 15 mM NaCI, 1 mM sodium phosphate, 0.2 mM Na2EDTA, pH 7.4, and dialyzed exhaustively against this buffer. It was then diluted with unlabelled DNA and the specific radioactivity determined from the DNA concentration estimated spectrophotometrically (taking A260 = 20 for 1 mg/ml) and the radioactivity determined by scintillation counting.…”

Section: Determination Of' Histone: Dna Rutiosmentioning

confidence: 99%

Differences in the binding of H1 variants to DNA

Clark

Thomas

1988

European Journal of Biochemistry

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A study of the complexes formed between short linear DNA and three H1 variants, a typical somatic HI, and the extreme variants H5, from chicken erythrocytes, and spHl from sea urchin sperm, has revealed differences between HI, H5 and spHl that have implications for chromatin structure and folding.1. All three histones bind cooperatively to DNA in 35 mM NaCl forming similar, but not identical, rod-like complexes. With sufficiently long DNA the complexes may be circular, circles forming more easily with H5 and spH1 than with H1.2. The binding of H5 and spHl to DNA is cooperative even in 5 mM NaC1, resulting in well-defined thin filaments that appear to contain two DNA molecules bridged by histone molecules. In contrast, H1 binds distributively over all the DNA molecules in 5 mM NaCl, but forms short stretches similar in appearance to the thin filaments formed with H5 and spH1. Rods appear to arise from the intertwining of regular thin filaments containing cooperatively bound histone molecules on raising the NaCl concentration to 35 mM.3. The compositions of the rods correspond to one histone molecule for about every 47 bp (Hl), 81 bp (H5) and 112 bp (spHl), suggesting average spacings of 24 bp (Hl), 41 bp (H5) and 56 bp (spH1) in the component thin (double) filaments. Strikinaly, these values are proportional to the linker lengths of the chromatins in which Histone the particular H1 variant is themain or sole H1.H1 serves the dual function of sealing the two turns of DNA around the histone octamer in the nucleosome and of mediating the ordered, salt-dependent folding of the 10-nm filament to form the 30-nm filament in vitro [I]. Different H1 variants may occur in different species, cell types and developmental stages [2] (and references therein). The structural significance of these variants is poorly understood [2], but it seems likely that they may be involved in the generation of folded filaments of different stabilities (cf.[3]), and that they may thereby play a role in regulating the accessibility of genes for transcription. Since all current models for the higher-order structure of chromatin [4] (and references therein) envisage the interaction of H1 to be primarily with DNA rather than with the core histones, we have asked whether there are differences in the interaction of different H1 variants with DNA that might account for the different properties in v i m and in vivo of the chromatins in which a particular variant is the sole, or major, HI.In an extension of earlier work on H1-DNA complexes [5], we have made a systematic study of the histone-DNA complexes formed by three quite distinct members of the H1 family: a typical somatic H1 (chicken Hl); the extreme variant, H5, which is the major lysine-rich histone in terminally repressed mature-chicken erythrocytes [6] ; and another extreme variant, spH1, from repressed sea-urchin sperm [7]. Chicken-erythrocyte chromatin and particularly sea-urchin-sperm chromatin are both highly condensed, and in addition to their distinctive H1 histones have long nucleosom...

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“…histone H I from sea urchin [44], capsid protein of Semliki Forest virus [45], and are thought to be involved in binding to nucleic acid. In the case of isolated histone HI, the C-terminal proline-lysine-alanine-rich region appears to possess little or no structure by 'H-NMR studies [46] but this region becomes fully immobilized upon interaction with DNA [47].…”

Section: Implications F O R the Role Of The Myosin Light-chain Isoenzmentioning

confidence: 99%

Proton Nuclear‐Magnetic‐Resonance Spectroscopy of Myosin Subfragment 1 Isoenzymes

Prince

Trayer

Henry

et al. 1981

European Journal of Biochemistry

103

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High-resolution proton N M R spectrocopy has been used to study the solution structures of the subfragment 1 (SI) isoenzymes (containing either the A1 or A2 light chains) from rabbit skeletal muscle myosin and to investigate their interaction with actin. Superimposed upon broad components, the narrow signals of the S1 spectra are unexpectedly sharp, indicating that domains of varying sidechain mobility occur in the conformation adopted in solution. These observations are in agreement with previous studies of the mixed isoenzymes [Highsmith et al. (1 979) Biochemistry, 18, 4238 -42431. Peptide amide exchange studies show also that the S 1 structure accommodates fluctuations of sufficient amplitude to allow most of the peptide groups to come into contact with the solvent on the time scale-of the 'H-NMR experiment. The overall impression is that S1 is a molecule possessing backbone motility as well as domains of different sidechain mobility.Close comparison of the Sl(A1) and Sl(A2) spectra indicate that the N-terminal41 residues of the A1 light chain, rich in lysine, proline and alanine, display a high degree of segmental mobility. The difference spectrum [SI(AI)-Sl(A2)] obtained closely resembles the spectral simulation of the 41-residue segment. Upon addition of actin, many of the narrow S1 resonances decrease in intensity or progressively disappear altogether, indicative of intermediateslow exchange conditions consistent with the recognised high affinity between the two proteins. These changes are interpreted as an overall modulation in the observed and hence more mobile regions of S1 as has been suggested in earlier H-NMR studies referred to above. In particular, the differences noted between S l(A 1 ) and S l(A2) have now largely disappeared in their complexes with actin indicating a marked reduction in the segmental mobility of the N-terminal region of the light chain in S l(A 1). Together with other affinity chromatography results [Winstanley and Trayer (1 979) Biochem. Soc. Trans. 7, 703 -7041, this is good evidence for a direct interaction between this area of S l(A 1) and actin.The mechanism of muscle contraction, as originally proposed by Huxley [I] and modified by several groups of authors [2, 31, consists of a cycle of cross-bridge detachments and attachments, which are the basis of the 'sliding filament' theory, The myosin heads form cross-bridges with actin, which are then detached by ATP. The hydrolysis of ATP produces some conformational change in myosin which is restored as mechanical energy when the heads rebind actin. There is now ample evidence that this theory is correct. There are, however, considerable differences of opinion with regard to the intermolecular and intramolecular interactions involved in the actomyosin complex, and several models have been proposed, as reviewed by Taylor [4]. X-ray diffraction and electron microscopy indicate that the cross-bridge or part of it rotates during contraction [5] and the existence of a 'swivel' and a 'hinge' have been postulated in myosin [6 -1 I...

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The Primary Structure of Histone H1 from Sperm of the Sea Urchin Parechinus angulosus. 2. Sequence of the C-Terminal CNBr Peptide and the Entire Primary Structure

Cited by 109 publications

References 12 publications

The amino acid sequence of wheat histone H2A(1). A core histone with a C-terminal extension

The amino acid sequence of wheat histone H2A(1). A core histone with a C-terminal extension

Differences in the binding of H1 variants to DNA

Proton Nuclear‐Magnetic‐Resonance Spectroscopy of Myosin Subfragment 1 Isoenzymes

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