共查询到20条相似文献,搜索用时 29 毫秒
1.
Robert C. Krueger 《Molecular biology reports》1986,11(3):189-192
Nucleosomal subunits isolated from rabbit thymus nuclei in 0.04 M K2SO4-0.02 M Tris, pH 7.4 were devoid of histone H1, while whole chromatin prepared in the same buffer contained the full complement of histone H1. The question is asked why histone H1 dissociates from the subunits but not from the high molecular weight material. We propose that, at physiological salt concentrations, histone H1 is not bound to linker DNA as depicted in the current models; rather, alternate attachment sites, present only in the polymer, are involved. 相似文献
2.
Shoulars K Brown T Alejandro MA Crowley J Markaverich BM 《Biochemical and biophysical research communications》2002,296(5):1083-1090
[3H]Luteolin binds covalently to uterine nuclear type II sites [B. Markaverich, K. Shoulars, M.A. Alejandro, T. Brown, Steroids 66 (2001) 707] and was used to identify this protein(s). SDS-PAGE analyses of [3H]luteolin-labeled type II site preparations revealed specific binding to 11- and 35-kDa proteins. The 11-kDa protein was identified as histone H4 by amino acid sequencing. Western blotting confirmed that the 11- and 35-kDa proteins were acetylated forms of histone H4. Anti-histone H4 antibodies (but not H2A, H2B, or H3 antibodies) quantitatively immunoadsorbed type II binding sites from nuclear extracts. Binding analyses by [3H]estradiol exchange, using luteolin as a competitor, detected specific type II binding activity to histone H4 (but not histones H2A, H2B, or H3) generated in a rabbit reticulocyte lysate translation system and confirmed that histone H4 is the type II site. 相似文献
3.
Analysis of Histones Associated with Neuronal and Glial Nuclei Exhibiting Divergent DNA Repeat Lengths 总被引:2,自引:1,他引:1
Abstract: Total cerebral hemisphere nuclei purified from adult rabbit brain were subfractionated into neuronal and glial populations. Previous studies have shown that chromatin in neuronal nuclei is organized in an unusual nucleosome conformation compared with glial or kidney nuclei, i.e., a short DNA repeat length is present. We now analyze whether this difference in chromatin organization is associated with an alteration in the histone component of nucleosomes. Total histone isolated by acid/urea-protamine extraction of purified neuronal, glial, and kidney nuclei was analyzed by electrophoresis on SDS-polyacrylamide slab gels. Histone H1 that was selectively extracted from nuclei was also examined. Differences were not observed on SDS gels in the electrophoretic mobilities of histones associated with either the nucleosome core particle (histones H2A, H2B, H3, H4) or the nucleosome linker region (histone H1). Total histone and selectively extracted histone H1 were also analyzed on acid/urea slab gels that resolve histones on the basis of both molecular weight and charge differences. When analyzed in this system, differences with respect to electrophoretic mobility were not detected when comparing either selectively extracted histone H1 or total histone from neuronal and glial nuclei. Quantitative analyses were also performed and neuronal nuclei were found to contain less histone H1 per milligram DNA compared with glial or kidney nuclei. Neuronal nuclei also demonstrated a lower ratio of histone H1/core histone. These results suggest that the pronounced difference in chromatin organization in neuronal compared with glial nuclei, which is reflected by a short DNA repeat length in neurons, appears to be associated with quantitative differences in neuronal histone H1. 相似文献
4.
A major rabbit skeletal muscle phosphorylase phosphatase activity which is markedly stimulated by histone H1 has been resolved from inhibitor-sensitive phosphorylase phosphatase (type-1 phosphatase), glycogen synthase kinase 3-activated phosphatase, phosphatase heat-stable inhibitor proteins, and alkaline phosphatase activity by various purification techniques. Evidence is presented that this phosphatase is a high-molecular weight form of a type-2 phosphatase. Our data suggest that this phosphatase may be regulated by histone H1, protamine or analogous polycationic compounds. 相似文献
5.
《The International journal of biochemistry》1991,23(2):189-194
- 1.1. The issue of the existence and the properties of histone H1° in quiescent rabbit tissues was approached by electrophoretic, Chromatographic and immunochemical techniques.
- 2.2. The results demonstrated that the rabbit tissues did contain a typical histone H1°, its properties being comparable to those of H1° from all other sources studied thus far.
6.
A chromosomal histone, H2S, specific to the mouse testis has been purified. Amino acid analysis indicated lack of cysteine and a high basic amino acid content typical of histones. Specific antibodies against histones H2S have been generated in rabbits and partially purified using (NH4)2SO4 precipitation and ion-exchange chromatography. Protein transfer experiments indicate presence of antigenically similar histones in the rat and rabbit testes but not in the guinea pig and dog testes. In addition, histone complement of somatic tissues such as lung, kidney, liver and spleen lacked antigenically similar proteins. Immunocytochemical studies using peroxidase-antiperoxidase complex indicated presence of immunoreactive cells in the seminiferous epithelium which were lacking in the interstitium. These data demonstrate histone H2S to be a unique histone associated with spermatogenesis in the mouse. 相似文献
7.
The relative quantity of histone H1 in rabbit thymus whole histone was determined to be 17.2% (w/w). This implies that there is, on average, one histone H1 molecule per nucleosome. 相似文献
8.
Availability of hyperacetylated H4 histone in intact nucleosomes to specific antibodies 总被引:4,自引:0,他引:4
Specific antibodies against the tetra-acetylated form of H4 histone have been elicited in the rabbit. They do not cross-react with the non-, mono-, and di-acetylated forms of the histone molecule but a slight cross-reactivity with the tri-acetylated form of H4 histone is observed. Our studies also show that hyperacetylated H4 histones are recognized by the antibodies in intact nucleosomes. 相似文献
9.
1. A calmodulin-binding protein of apparent mol.wt. 19 000 has been purified from chicken gizzard. Similar proteins have been isolated from bovine uterus, rabbit skeletal muscle and rabbit liver. 2. These proteins migrated as an equimolar complex with bovine brain calmodulin on electroporesis on polyacrylamide gels in the presence of Ca2+ and 6M-urea. The complex was dissociated in the presence of EGTA. 2. The chicken gizzard calmodulin-binding protein has been shown to be identical with chicken erythrocyte histone H2B on the basis of partial amino acid sequence determination. 4. The calmodulin-binding proteins of apparent mol.wt. 22 000 isolated previously from bovine brain [Grand & Perry (1979) Biochem. J. 183, 285-295] has been shown, on the basis of partial amino-acid-sequence determination, to be identical with myelin basic protein. 5. The activation of bovine brain phosphodiesterase by calmodulin is inhibited by excess bovine uterus calmodulin-binding protein (histone H2B). 6. The phosphorylation of myelin basic protein by phosphorylase kinase is partially inhibited, whereas the phosphorylation of uterus calmodulin-binding protein (histone H2B) is unaffected by calmodulin or troponin C. 7. The subcellular distribution of myelin basic protein and calmodulin suggests that the two proteins do not exist as a complex in vivo. 相似文献
10.
A solid-phase radioimmunoassay allows detection of small structural differences in histones. In this assay, histones are absorbed to plastic tubes; the coated tubes bind antibody from the IgG fraction of antihistone rabbit antiserum, and the bound rabbit IgG is detected with 125I-labeled purified goat anti-rabbit IgG. H2a-anti-H2a and H2b-anti-H2b systems showed no cross-reactivity with each other. H2a variants (H2a.1 and H2a.2) showed some cross-reaction but were distinguished reciprocally by corresponding antisera even though they differ in only two amino acids. This quantitative distinction was detected with a wide range of IgG concentrations, whereas only low concentrations revealed the differences in complement fixation assays. Histones in solution competed with the tube-absorbed histone for binding of the IgG. 相似文献
11.
A histone lysine methyltransferase activated by non-canonical Wnt signalling suppresses PPAR-gamma transactivation 总被引:4,自引:0,他引:4
Takada I Mihara M Suzawa M Ohtake F Kobayashi S Igarashi M Youn MY Takeyama K Nakamura T Mezaki Y Takezawa S Yogiashi Y Kitagawa H Yamada G Takada S Minami Y Shibuya H Matsumoto K Kato S 《Nature cell biology》2007,9(11):1273-1285
12.
Y.M. Bhatnagar K. McCullar R.D. Faulkner R.D. Ghai 《Biochimica et Biophysica Acta (BBA)/General Subjects》1983,760(1):25-33
A chromosomal histone, H2S, specific to the mouse testis has been purified. Amino acid analysis indicated lack of cysteine and a high basic amino acid content typical of histones. Specific antibodies against histones H2S have been generated in rabbits and partially purified using (NH4)2SO4 precipitation and ion-exchange chromatography. Protein transfer experiments indicate presence of antigenically similar histones in the rat and rabbit testes but not in the guinea pig and dog testes. In addition, histone complement of somatic tissues such as lung, kidney, liver and spleen lacked antigenically similar proteins. Immunocytochemical studies using peroxidase-antiperoxidase complex indicated presence of immunoreactive cells in the seminiferous epithelium which were lacking in the interstitium. These data demonstrate histone H2S to be a unique histone associated with spermatogenesis in the mouse. 相似文献
13.
Anna Akhmanova Koos Miedema Yaxian Wang Mieke van Bruggen Jo H. M. Berden Evangelos N. Moudrianakis Wolfgang Hennig 《Chromosoma》1997,106(6):335-347
A rabbit antiserum, specific for the histone H3.3 replacement variant, was raised with the aid of a histone H3.3-specific
peptide. Immuno blot experiments demonstrated the specificity of this polyclonal antiserum. In addition, we showed on immuno
blots that two monoclonal antibodies isolated from mice with systemic lupus erythematosus (SLE) display strong reactivity
with the H3.3 histone, but not with its replication-dependent counterparts. Our observations indicate that histone H3.3 might
play a role as autoantigen in SLE. We used the histone H3.3-specific antiserum to characterize the germ line chromatin in
cytological preparations of Drosophila testes, because our previous studies had shown that a histone H3.3-encoding gene is strongly expressed in the germ line of
Drosophila males. The antiserum reacted with some of the lampbrush loops in spermatocytes and with chromatin of the postmeiotic germ
cells of males. Our data indicate that histone H3.3 is not evenly distributed throughout the chromatin of germ cells, but
is concentrated in distinct regions. Histone H3.3 disappears from the spermatid nuclei, along with the other core histones,
during the late stages of spermatogenesis. In Drosophila polytene chromosomes, however, a rather uniform distribution of the histone H3.3 was observed. The possible role of histone
H3.3 is discussed.
Received: 12 May 1997 / Accepted: 4 July 1997 相似文献
14.
We studied the expression of the osteogenic and antinociceptive C-terminal histone H4-related peptide fragments, H4-(89-102) (OGP) and H4-(86-100), respectively, within various rat tissues and isolated alveolar macrophages (AM) by radioimmunoassay (RIA). OGP was located mainly within the bone marrow, spleen, thymus, and lungs whereas H4-(86-100) was more concentrated within the bone marrow, lymph nodes, spinal cord, pituitaries and thymus. The expression pattern of the two peptides showed similarities with the tissue expression pattern of the histone H4 mRNA variant H4-v.1. In rat AM, OGP and H4-(86-100) levels were significantly stimulated (2.6- and 1.9-fold, respectively) by LPS (1 microg/ml), along with H4-v.1 mRNA (4.1-fold), but not whole histone H4 (1.1-fold) nor total histone H4 mRNA (1.1-fold). The results suggest that H4-v.1 mRNA may play a role in the synthesis of the naturally occurring peptides H4-(86-100) and OGP via the alternative translation product H4-(84-102), but not whole histone H4. 相似文献
15.
To better understand the basis for heat shock-induced chromatin condensation in Achlya, a further characterization of the histones of this organism was carried out. The nucleosomal location (i.e., core vs linker), partial peptide map, and electrophoretic behavior of each Achlya histone was determined and compared to the well-characterized histones of rabbit kidney. The results of this and previous studies suggest that in Achlya, no nucleosome linker-associated histone analogous to histone H1 of higher eucaryotes is observed and that the Achlya histone designated alpha is a novel nucleosomal core histone. These observations may reflect the existence of a mechanism of stress-induced chromatin condensation which does not involve histone H1. 相似文献
16.
Influence of histone phosphorylation upon histone-histone interactions studied in vitro 总被引:1,自引:0,他引:1
Histones H2b and H3, phosphorylated in vitro with the catalytic subunit of protein kinase I from rabbit skeletal muscle, were used to estimate the influence of histone phosphorylation upon histone-histone complex formation. Stoichiometry and interaction affinity of the complexes H2a-H2b, H4-H2b, and H4-H3 were determined by using the continuous variation method based on circular dichorism or fluorescence intensity. All complexes exhibit a 1:1 stoichiometry in sodium phosphate or sodium chloride solution of pH 7.0. The association constants of the complexes containing phosphorylated H2b were only slightly reduced, whereas that with phosphorylated H3 was strongly reduced relative to those of the nonphosphorylated species. 相似文献
17.
Histone H2AX and Fanconi anemia FANCD2 function in the same pathway to maintain chromosome stability
Bogliolo M Lyakhovich A Callén E Castellà M Cappelli E Ramírez MJ Creus A Marcos R Kalb R Neveling K Schindler D Surrallés J 《The EMBO journal》2007,26(5):1340-1351
Fanconi anemia (FA) is a chromosome fragility syndrome characterized by bone marrow failure and cancer susceptibility. The central FA protein FANCD2 is known to relocate to chromatin upon DNA damage in a poorly understood process. Here, we have induced subnuclear accumulation of DNA damage to prove that histone H2AX is a novel component of the FA/BRCA pathway in response to stalled replication forks. Analyses of cells from H2AX knockout mice or expressing a nonphosphorylable H2AX (H2AX(S136A/S139A)) indicate that phosphorylated H2AX (gammaH2AX) is required for recruiting FANCD2 to chromatin at stalled replication forks. FANCD2 binding to gammaH2AX is BRCA1-dependent and cells deficient or depleted of H2AX show an FA-like phenotype, including an excess of chromatid-type chromosomal aberrations and hypersensitivity to MMC. This MMC hypersensitivity of H2AX-deficient cells is not further increased by depleting FANCD2, indicating that H2AX and FANCD2 function in the same pathway in response to DNA damage-induced replication blockage. Consequently, histone H2AX is functionally connected to the FA/BRCA pathway to resolve stalled replication forks and prevent chromosome instability. 相似文献
18.
Antiserum prepared from a rabbit which was immunized with human erythrocyte glycerate-2,3-P2 synthase was found to react with glycerate-2,3-P2 synthase in rabbit erythroid cells. By using this antiserum, it was proved that the specific activity of this enzyme was unchanged during the development of the rabbit erythroid cells. This leads us to conclude that the increased activity of glycerate-2,3-P2 synthase in developing erythroid cells (Narita, H., Ikura, K., Yanagawa, S., Sasaki, R., Chiba, H., Saimyoji, H., and Kumagai, N. (1980) J. Biol. Chem. 255, 5230-5235) is due to the accumulation of enzyme protein. There is at least a 16-fold increase in the level of this protein during development from bone marrow erythroid cells to erythrocytes. The synthesis of glycerate-2,3-P2 synthase was shown to occur in rabbit reticulocytes and bone marrow erythroid cells. These cells were incubated for protein synthesis and the protein synthesized was precipitated with the anti-glycerate-2,3-P2 synthase antiserum and separated on sodium dodecyl sulfate-polyacrylamide gels. The immunoprecipitated product was shown to produce fragments of the same molecular weight after digestion with V8 protease as did the pure glycerate-2,3-P2 synthase. The proportion of glycerate-2,3-P2 synthase synthesis in reticulocytes (0.04% of total protein synthesis) was comparable to the level of this protein in the cells (0.07% of the total protein). 相似文献
19.
Chromatin condensation and histone H1 kinase activity during growth and maturation of rabbit oocytes
Ladislava Jelíankov Michal Kubelka Jan Motlík Pierre Guerrier 《Molecular reproduction and development》1994,37(2):210-215
Fully grown rabbit oocytes, isolated from preovulatory follicles, exhibit highly condensed bivalents within an intact germinal vesicle while a very low level of histone H1 kinase activity could be detected in their extracts. Chromatin condensation started in growing oocytes isolated from antral follicles presenting a diameter of 0.5 mm. This event was accompanied by a transient rise in histone H1 kinase activity which culminated in large antral follicles measuring 0.75 to 1 mm in diameter. However, the extent of histone H1 kinase activity observed in these growing oocytes remained far less important than that recorded in extracts prepared from in vitro cultured metaphase I and metaphase II oocytes. Moreover, this activity was insufficient to induce germinal vesicle breakdown which will only occur with an increasing efficiency, following in vitro culture of medium, large, and fully grown antral follicles. © 1994 Wiley-Liss, Inc. 相似文献