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The appearance of methylated lysines in newly synthesized histones from Ehrlich ascites tumor cells was measured during one generation time. Newly synthesized histones were pulse-labeled in vivo by L-[3H]lysine, and the time course of the uptake of label into monomethyl, dimethyl and trimethyllysine from gel-electrophoretically isolated histones F2a1 (H4) and F3 (H3) was followed. Methylation starts immediately after histone biosynthesis. It proceeds, however, more slowly than histone synthesis. Both the rate of methylation and the mechanism of methylation in F3 and F2a1 histones differ. F3 methylation can be described by a first-order reaction, i.e. the reaction rate depends only on the concentration of free methylation sites available. Rate constants of approximately 0.21 h-1 were found for all three methylation steps. Methylation in the F2a1 histone proceeds more slowly than in F3. The dimethylation step in this fraction can be described by a zero-order reaction with a rate constant which is the reciprocal of the duration of the DNA synthesis phase. Alternatively this step could be correlated with the transition of the cells from the S phase into the G2 phase. By the end of one generation time all methylation sites in all F2a1 and F3 molecules are occupied by methyl groups at a ratio of about 1:3:1 for monomethyl, dimethyl and trimethyllysine in the F3 histone. In the F2a1 molecule the methyllysines consist mainly of dimethyllysine. 相似文献
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M A Sibileva T N Osipova A O Zalenski? M A Chebishian A I Golikova 《Molekuliarnaia biologiia》1976,10(2):514-520
Characteristic viscosity, sedimentation constant and optical anisotropy were studied of the complexes formed between DNA and histone fractions F3 and F3+F2a2. The parameters mentioned continuously change with the increase of protein content within the complex. Analysis of experimental data shows that binding of a histone bads to a decrease of size and thermodynamic rigidity of the DNA molecule. On the basis of results obtained a model of F3 histone binding with DNA is suggested, amino acid sequence of this protein being taken into account. Comparison of behaviour of nucleohistones DNA+F3 and DNA+F1 studied previously testifies different way of binding of these histones to DNA. 相似文献
4.
1. The interactions of the lysine-rich histone F1 with DNA have been studied at various histone to DNA ratios, in water and in the presence of uni- and bi-valent cations. In water only, histone F1, even in fourfold excess, is unable to precipitate all the DNA. In 0.14m-sodium chloride, 0.8mg. of histone F1 is required to precipitate 1mg. of DNA, whereas in 0.07m-magnesium chloride only 0.4mg. is required. 2. Bivalent cations are also shown to be more effective in dissociating the DNA-histone complex. Histone F1 can be selectively removed from deoxyribonucleoprotein with 0.1m-magnesium chloride. 3. The precipitation of DNA by histone F1 is a reversible process and the complex can be taken in and out of solution by changing the ionic environment. 4. The bearing of these results on the observed ability of various DNA-histone complexes to act as templates for RNA synthesis is discussed. 相似文献
5.
Comparison between histones FV and F2a2 of chicken erythrocyte. II. Interaction with homologous DNA.
The conformation and stability of artificial complexes between chicken erythrocyte DNA and homologous histones FV and F2a2 was studied by circular dichroism (CD) and thermal denaturation followed by both absorbance and CD measurements. The complexes are made after a stepwise potassium fluoride gradient dialysis without urea and studied at low ionic strength (10-minus 3 M). 1) No structural changes of the DNA can be detected up to r equals 0.2 with FV and r equals 0.6 for F2a2. With FV at higher values of r the CD spectrum is altered, indicating the organization of DNA and histones in some kind of aggregate. 2) The conformation of histone molecules inside the complexes is not related to the ionic strength of the medium but to an effective ionic environment close to 0.1 M. This ionic strength would also correspond to the melting temperature of histone-covered DNA. 3) From the analysis of the absorbance melting profile the length of DNA covered with an histone molecule can be estimated. A good agreement is found between the negative charge of this piece of DNA and the net positive charge of the histone. 4) Since the CD transition at 227 nm occurs before the second absorbance transition at 280 nm, the DNA is stabilized no longer by native histone but partially or fully denatured histones. The helical regions of the histone molecule are not involved in the binding process, which appears to be almost purely coulombian and most likely related to some structural fit between the pattern of negative charges in the DNA helix and that of positive charges along the peptide chain. 相似文献
6.
Variations in the phosphate content and thiol/disulphide ratio of histones during the cell cycle. Studies with regenerating rat liver and sea urchins 总被引:5,自引:5,他引:0
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1. In regenerating rat liver the phosphate content of the lysine-rich histone F1, but not that of the more arginine-rich histone F3-1, increases during the period of DNA synthesis. 2. The phosphorylation of histone F1 in this ;S period' is decreased by gamma-irradiation, but, like phosphate uptake into DNA, is affected to an even greater extent if the irradiation is given in the presynthetic period. 3. Histones from three species of sea-urchin eggs show similarities to the F2 and F3 groups of histones from mammalian thymus gland. 4. The proportion of thiol to total thiol plus disulphide in acid extracts from sea-urchin eggs varies from less than 20% in mature unfertilized eggs to 59% just before cleavage. 5. The phosphorylated forms of histones F1 and F3 are less effective in decreasing DNA synthesis by DNA polymerase than the non-phosphorylated forms. 6. Oxidation of thiol groups on histone F3-1 does not affect its capacity to decrease DNA synthesis in vitro. 相似文献
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Chemical and metabolic properties of adenosine diphosphate ribose derivatives of nuclear proteins. 总被引:3,自引:0,他引:3
1. ADP-ribose is found in rat liver nuclei covalently bound to histone F1, to a non-histone protein, and to a small peptide. 2. A single unit of ADP-ribose, covalently bound to phosphoserine, was isolated from an enzymic hydrolysate of histone F1. ADP-ribose-bearing peptides were isolated from a tryptic digest of the histone. 3. It is proposed that the 1'-hydroxyl group of ADP-ribose is linked to the phosphate group of phosphoserine in histone F1. 4. The incorporation of 32P into ADP-ribose on histone F1 a parallels the DNA content through the cell cycle. An increased incorporation of the nucleotide into the other derivatives is observed during S phase. 5. It is suggested that the ADP-ribose derivative of histone F1 has a role in maintaining the G0 state and that one or both of the other derivatives is concerned with control of DNA synthesis. 相似文献
8.
Variations in the phosphate content of histone F1 in normal and irradiated tissues 总被引:13,自引:10,他引:3
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1. The capacity of the histone-DNA complex to act as a template for RNA synthesis is increased by phosphorylation of histone F1. 2. In regenerating liver, DNA synthesis is preceded by phosphorylation of histone F1. 3. Exposure to ionizing radiation in vivo decreases and delays the phosphorylation of histone F1 in regenerating liver, and decreases it in the hyperplastic kidney. 4. Histone F1 is phosphorylated to a greater extent in dividing than in resting tissues. 相似文献
9.
1. Two minor protein fractions (B and C) were separated from histone F1 by chromatography on DEAE-cellulose. Fraction B was acidic. 2. Uptake of (32)P in vivo into histone F1 but not into fractions B and C was stimulated by partial hepatectomy. 3. It is suggested that partial hepatectomy causes an increase in the number of histone F1 molecules phosphorylated. 相似文献
10.
Interactions of calf-thymus histone fractions in aqueous solution with 8-anilinonaphthalene-1-sulphonic acid 总被引:4,自引:3,他引:1
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D J Laurence 《The Biochemical journal》1966,99(2):419-426
1. The interactions of histone fractions with 8-anilinonaphthalene-1-sulphonic acid were investigated by fluorimetry and spectrofluorimetry and the results were interpreted with the aid of equilibrium-dialysis techniques. 2. Characteristic differences were found between the various histone fractions, and with fractions F3 and F2a the binding was found to be salt-dependent. 3. Evidence was obtained indicating a slow change of the physical state of fractions F3 and F2a in the presence of salt, and the binding by these two fractions in the presence of salt was greater by an order of magnitude than by fractions F1 and F2b. 4. Conditions favouring binding were also those favouring histone aggregation; SO(4) (2-) ions activated binding at a lower concentration than Cl(-) ions; urea, guanidinium ions and high concentrations of I(-) ions were inhibitory to binding. 5. After histones had been kept in the presence of salt for a long time the reversal of interaction on decreasing the salt concentration was incomplete. 6. The inhibition of binding by fraction F2a in the presence of urea or fraction F2b depended on the time sequence of addition of the reagents. 7. Artificial nucleoproteins made by precipitating DNA with the histone fractions in neutral 0.14m-sodium chloride showed the same order of interaction as was found for the fractions in solution. 8. Comparison of the binding by fraction F2a with that by bovine plasma albumin showed that in both cases there were a large number of weakly binding sites but that fraction F2a lacked the small number of strongly binding sites found in albumin. No slow change of binding in the presence of salt was found for albumin. 9. Binding by fraction F2b increased the affinity of the protein for further molecules of the adsorbate. 10. The results are discussed in relation to the close relationship between binding and aggregation and the possible role of non-polar interactions as determined by the balance between polar and non-polar amino acids in the histone fractions. 相似文献
11.
1. The activity of the soluble phosphokinase for histone F1 increases in regenerating rat liver during the first period of DNA synthesis after partial hepatectomy. The increase probably represents new enzyme synthesis. 2. A dose of 500rd of gamma-irradiation given early in G1 decreases the amount of histone F1 phosphokinase found 22h after partial hepatectomy by 60-70%. 3. The enzyme preparations also contained a histone F1 phosphatase; the presence together of the kinase and phosphatase caused a disproportion between net (31)P uptake and (32)P incorporation into histone F1. 4. All four subclasses of histone F1 could accept phosphate from ATP. 5. Crude enzyme preparations transferred more (31)P into histone F1 with an initially low phosphate content than into one with a high phosphate content; conversely, more (32)P was transferred into the latter. 相似文献
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The triiodothyronine (T3) nuclear receptor was previously shown to lose rapidly its high affinity hormone-binding property after a partial purification from the nuclear extract. It was then found that histones + DNA added to the incubation medium with labeled T3 could restore, at least in part, the high affinity T3 binding. We now demonstrate that DNA alone increases the high affinity T3 binding site concentration moderately, and only at low ionic strength where it can bind to the receptor. Total histones and all histone fractions studied (total core histones, F2a, H2B, H3, H4, H1) specifically increase, at low concentrations, the level of T3 binding; but higher concentrations of some individualized histones, particularly arginine-rich histones, have an inhibitory effect. DNA, or several other polynucleotides, in the presence of histones increase the stimulating histone effect and reverse the inhibitory effect into a true activation. Histones increase the number of T3 binding sites but decrease the affinity for T3; addition of DNA restores the high affinity for T3 and stabilizes the T3-receptor complexes. Thus, some of the histone molecules could play a role in the maintenance of the T3 binding site, but multiple interactions between histones or with DNA seem necessary to impair the negative effect exerted by other parts of the histone molecules. Whether these positive and negative effects of histones on the T3 binding site are of biological relevance in the regulation of T3 binding to its receptor remains to be determined. 相似文献
14.
Lysine-rich histone isolated from different chicken tissues was separated electrophoretically into 4-5 subfractions. The subfrations reffered to as 1, 2, 3, and 4, occur in each the tissue studied, erythrocyte lysine-rich histone containing an additional subfraction 1a. F1 histone from mitotically active tissues (intestinal mucosa, thymus, testes) has a higher content of subfraction 2, while the same histones from mototically inactive tissues (liver, heart, brain) contain an elevated amount of subfraction 3. F1 histone isolated from liver, brain and heart of 21-day embryo has much more of subfraction 2, than the same histone of adult animal. During the chicken development from hatching till maturation the content of subfraction 2 in these organs decreases, and the content of subfraction 3 increases. The rate of this change in liver corresponds to the rate of DNA synthesis. In F1 histone of erythrocytes the content of subfraction 4 falls down during the post hatching ontogenesis. 相似文献
15.
R. J. Inglis T. A. Langan H. R. Matthews D. G. Hardie E. M. Bradbury 《Experimental cell research》1976,97(2):418-425
The previous observation that growth-associated histone kinase (HKG) from Ehrlich ascites cells brings forward mitosis in Physarum polycephalum has been confirmed with more step 1 histone kinase and a more purified (step 2) histone kinase and the statistical significance of the results assessed. The mitosis appears normal in the phase contrast microscope and DNA synthesis is initiated after mitosis as usual. In vitro the growth-associated histone kinase phosphorylates chromatin, the phosphate appearing in F 1 histone. The results are interpreted as providing support for the hypothesis that growth-associated histone kinase controls the initiation of mitosis through F 1 histone phosphorylation and chromosome condensation. 相似文献
16.
The kinetics of nitration of tyrosine residues in histones F1 and F2a1 by tetranitromethane has been investigated. At low ionic strength and 30-fold molar excess of nitrating agent the nitration reaction results in fast modification of all tyrosine residues in both histones. At the same time the rates of modification of different tyrosine residues in histone F2a1 are not identical and markedly exceed the rate of N-Ac-OEt-Tyr nitration in a model system. The increase of reaction mixture ionic strength causes an increase of modification rates. The differential UV-absorption spectra of histone F1 obtained by temperature perturbation show an abnormal positive characteristic maximum at 286.8 nm. Analysis of the dependence of nitration rates of tyrosine residues in histones in saline solutions upon the ionic strength and of difference UV-absorption spectra of histones leads to a conclusion that there are specific interactions of definite parts of histone polypeptide chains. These interactions may arise from aggregation of histone molecules. 相似文献
17.
Electron microscopic and biochemical evidence that chromatin structure is a repeating unit 总被引:1,自引:0,他引:1
Electron microscopic and biochemical studies demonstrate that the fundamental structure of chromatin depleted of lysine-rich histones is composed of a flexible chain of spherical particles (nucleosomes), about 125 Å in diameter, connected by DNA filaments. Such a chromatin preparation can be separated by centrifugation into two fractions which differ in the spacing of the nucleosomes. In one fraction almost all of the DNA is condensed in nucleosomes, while the other fraction contains long stretches of free DNA connecting regions where the nucleosomes are closely packed. The isolated nucleosomes contain about 200 base pairs of DNA and the four histones F2a1, F2a2, and F2b, and F3 in an overall histone/DNA ratio of 0.97. In such a structure the DNA is compacted slightly more than five times from its extended length. The same basic structure can be visualized in chromatin spilling out of lysed nuclei. However, in this latter case the nucleosomes are very closely packed, suggesting that histone F1 is involved in the superpacking of DNA in chromosomes and nuclei. The chromatin fiber appears to be a self-assembling structure, since the nucleosomal arrangement can be reconstituted in vitro from DNA and the four histones F2a1, F2a2, F2b and F3 only, irrespective of their cellular origin. 相似文献
18.
The synthesis and decay of histone fractions and of deoxyribonucleic acid in the developing avian brain
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Stephen C. Bondy 《The Biochemical journal》1971,123(3):465-469
1. The turnover of cerebral histones and DNA after injection of [4,5-(3)H]leucine or [methyl-3-(3)H]thymidine, respectively, was studied in the developing chick. 2. Chromatin was prepared from chick nuclei that had been purified by centrifugation through 1.9m-sucrose. 3. Nuclear proteins were fractionated into three major histone classes, F1 (lysine-rich), F2(b) (slightly lysine-rich) and [F3+F2(a)] (arginine-rich), and a non-histone protein residue. 4. The proportions of the histone classes remained constant throughout the period of development studied. 5. All histone fractions decayed at a similar rate, initially with a half-life of around 5 days, later with a half-life of 19 days. 6. Non-histone proteins from chromatin decayed in a heterogeneous manner with a wide range of half-lives. 7. Short-term labelling studies showed that all histone fractions were synthesized at the same rate. 8. Some non-histone proteins were very rapidly synthesized relative to histones. 9. DNA had a longer half-life than any histone fraction studied. A biphasic exponential decay curve with half-lives of 23 and 50 days was found. 10. It was concluded that the turnover of histones can occur independently of that of DNA and that different histone classes have similar rates of synthesis and decay. 相似文献
19.
The gel-filtration behaviour of calf thymus histone fraction F2(b) was studied at three different salt concentrations (0.01m-, 0.10m- and 1.00m-sodium chloride) and two different pH ranges (pH3–4 and pH6.7–7.1). Other histone fractions [F1, F2(a) and F3] were also utilized to assist interpretation of the data. It was found that the Stokes radius of histone fraction F2(b) was not significantly changed when the salt concentration was increased, implying that the aggregation of the individual histone molecules (Edwards & Shooter, 1969) resulted in only relatively minor changes in the hydrodynamic volume. Aggregation would appear to be due to the salting out of hydrophobic regions giving rise, in the aggregate, to a compact core of hydrophobic groups from which protrude the remaining basic parts of the molecule. Repulsion between charged groups on the basic regions of individual histone molecules would give the aggregate approximately spherical symmetry, the diameter of the aggregate approximating to the length of a single histone molecule. 相似文献
20.
Y Yoshikawa Y S Velichko Y Ichiba K Yoshikawa 《European journal of biochemistry》2001,268(9):2593-2599
We report that complexes of giant DNA molecules with histone H1 proteins form a pearl necklace-like structure when the complexes are prepared by natural dilution from a high-salt solution (2 M NaCl) to a low-salt solution (0.2 M and 50 mM NaCl). We performed real-time observations on the conformational changes of individual T4 phage DNA (166 kb) molecules in bulk solution by fluorescence microscopy. To identify H1-binding regions on individual DNA molecules, we also performed immunofluorescence microscopic observations on the DNA-H1 complex spread on a glass surface. It was found that histone H1 binds DNA in a highly co-operative manner and is accompanied by local folding of the DNA. On the basis of the experimental observations and a theoretical simulation, we propose a self-assembling mechanism for the pearling structure. 相似文献