Search PubMed⌕ Search

Biomedical subjects

G I Chikhirzhina

Publications and source records attributed to G I Chikhirzhina.

14 recordsLinked to original sources

[Interaction of the regulatory region of the tryptophan oxygenase gene with transcription factors of the nuclear factor 1 (NF1) family].

Inducible hormone-dependent tryptophan oxygenase gene is expressed mostly in the liver under the control of glucocorticoid hormones. In the regulatory region of this gene there are three constitutive sites independent of hormone presence and gene expression. While investigating transcription factors responsible for the formation of specific chromatin structure in the regulatory region of inducible genes it is necessary to identify proteins which can bind to DNA specifically in these sites. The present paper reports investigation of transcription factors which bind to DNA from the -292nd to the -178th nucleotide of gene to in vitro. This DNA region contains a site corresponding to the constitutive DNase I hypersensitive site in vivo. Using electrophoretic mobility shift assay, we have analysed binding of rat liver proteins to this DNA region. NF1-rich nuclear protein fraction was purified from the rat liver nuclear extract by DEAE-cellulose and heparin-sepharose chromatography. Studies of the competition with a consensus sequence site for NF1 recognition sites have shown that it is the NF1 family transcription factors that are responsible for the formation of these specific complexes.

Animals↗

[Nuclear proteins, specifically binding the regulatory region of rat tryptophan oxygenase].

Using electrophoretic mobility shift assay we have analysed the binding of rat liver nuclear proteins to the fragments from -466 to -292 and from -292 to -178 relative to the transcriptional start site of the rat tryptophan oxygenase gene. Studies of the competition with a synthetic consensus sequence for the NF 1 recognition site have shown that the liver nuclear proteins, responsible for the formation of specific complexes with these fragments belong to the family of the nuclear factor 1 (NF 1). We have also found that the trans-acting factors of the NF 1 family form several complexes with each of the two tested fragments of the tryptophan oxygenase gene.

Animals↗

[Acid-soluble, non-histone proteins in rat liver chromatin. Interaction with DNA].

Some properties of nonhistone proteins of rat liver chromatin (Mr 40 +/- 1 and 41 +/- 1 KD) are described. These proteins are abundant in monomeric particles formed at the early steps of chromatin fragmentation by Ca2+,Mg2+-DNase. The proteins are not extracted from chromatin by 5% HClO4 and 1 M NaCl, but can be extracted by 0.4 n H2SO4 and 2 M NaCl. Study on proteins binding to DNA demonstrated that in 0.05 M NaCl these proteins are bound both to bovine satellite DNA and to the plasmid pBR 322 DNA.

Animals↗

[Independence of nucleolysis of structural elements of chromatin of histone H1].

The type of nucleolysis and the size of DNA monomeric particles were studied at different steps of rat liver chromatin digestion by endogenous Ca2+, Mg2+-DNAase. At an early stage of nucleolysis (5-7% of monomers, 1-2% of acid-soluble material) the monomeric particles are represented by three electrophoretic classes containing all histones and differing in DNA size; the latter varies from 170 to 215 base pairs. The monomers with histone H1 and without it isolated at the stage of exhaustive fragmentation (85-90% of monomers, 12-15% of acid-soluble material) are represented by two electrophoretic classes. The mobility of these particles does not depend on the presence of histone H1 in them. A comparison of DNA sizes in the particles with and without histone H1 demonstrated that these sizes are identical and vary from 150 to 175 base pairs. The formation of histone H1-free monomers is not accompanied by the increase in exonucleolytic degradation of chromatin DNA. A comparison of electrophoretic patterns of monomeric particle DNA at different steps of nucleolysis showed that degradation of DNA in the structural chromatin unit occurs uninterruptedly from 215 to 150 nucleotides and in the region of 175-150 nucleotides is independent of histone H1. The data obtained suggest that histone H1 does not protect the DNA against nuclelysis at nucleotides 150-175 of the chromatin structural unit, which is in agreement with the existing concepts on protection of this nucleosomal region by histone octamer.

Animals↗

[Fragmentation of chromatin by endogenous nucleases, accumulation of the subnucleosomal fragments with high electrophoretic mobility].

Digestion of chromatin by endogenous nucleases to nucleosomes (140-160 base pairs of DNA) is accompanied by the accumulation of subnucleosomal DNP particles with high electrophoretic mobility (20-40 base pairs of DNA). All histones associate with the 140-160 base pairs fragment. The production of subnucleosomal DNP particles does not correlate with the degradation of histone H1 and the appearance of nucleosomes lacking histone H1. Degradation of the protein in this fragment is accompanied by the appearance of free DNA. The data obtained are in agreement with the hypothesis on the origin of subnucleosomes from the nucleosomal locus preferentially associated with the non-histone proteins and on the autonomy of these loci and of the loci associated with histone H1 in the nucleosome.

Chromatin↗

[Solubilization of chromatin by an endogenous enzymic Ca2+, Mg2+-dependent factor. Activity of residual chromatin].

An endogenous Ca2+, Mg2+-dependent factor of enzymic nature (apparently an endonuclease) digests a part of chromatin in the rat liver nuclei producing DNA fragments of an uniform size. After 60 min of incubation at 15 degrees C and pH 7.50 in the presence of 5 mM MgCl2 and 2 mM CaCl2 87-93% of the total chromatin becomes soluble. The insoluble chromatin however contains 70-85% of the in vivo newly synthesized RNA. In regenerating liver the proportion of the insoluble residual chromatin increases while the radioactivity of the newly synthesized DNA in this fraction is highest. Residual chromatin can be solubilized by ultrasonic treatment only. The Ca2+, Mg2+-dependent dissolving factor is not present either in brain or in PMN leucocyte nuclei.

Animals↗

[Transcriptional regulation of placental transforming growth factor-beta by calcitriol in prostate cancer cells is androgen-independent].

Calcitriol (1 alpha,25-dihydroxycholecalciferol) inhibits prostate cancer cell growth. It has been shown that inhibition of growth of prostate cancer LNCaP cells by calcitriol is androgen-dependent. Using cDNA microarray we showed that calcitriol induced expression of placental transforming growth factor-beta (PTGF-beta), which is known to suppress cell growth. We studied regulation of PTGF-beta gene expression by calcitriol and 5alpha-dihydrotestosterone and analyzed whether induction of PTGF-beta transcription by calcitriol is androgen-dependent. Using real-time PCR we demonstrate that 5alpha-dihydrotestosterone up-regulates PTGF-beta mRNA. We do not find an effect of 5alpha-dihydrotestosterone or antiandrogen Casodex on calcitriol-induced PTGF-beta mRNA level and conclude that induction of PTGF-beta transcription by calcitriol is androgen-independent.

5-alpha-Dihydroprogesterone↗

[Distribution of transcription-active DNA sequences in rat liver chromatin during fragmentation with endogenous DNAses].

Distribution of the transcriptionally active sequences in rat liver chromatin DNA fragments released from chromatin sites with different sensitivity to endogenous Ca2+, Mg2+-DNases was studied by the dot hybridization method with cloned DNA-probes. The internal fragment of the rat chromosomal ceruloplasmin gene containing coding sequences and the promoter region of the metallothionein-I gene were specific probes for the transcriptionally active liver chromatin. cDNA of beta-globin gene was the control for the transcriptionally inert DNA sequences. Mononucleosomal length, DNA (175-215 bp) formed in the course of mile nuclease digestion (less than 1% mononucleosomes and acid-soluble material) was 3-6-fold enriched in transcribed sequences of the ceruloplasmin gene as compared to oligonucleosomal DNA fragments produced in the same digestion conditions (fractions containing fragments 750-850 and greater than 2000 bp). The relative amount of the ceruloplasmin gene in mononucleosomal length DNA formed during the mild digestion was 12-25-fold greater than in total rat liver DNA and 25-50-fold greater than in the mononucleosomal DNA (150-175 bp) produced in the course of extensive digestion (80-85% mononucleosomes and 15-20% acid-soluble material) by endogenous Ca2+, Mg2+-DNases. The promoter metallothionein-I gene sequences exhibited the same distribution among chromatin fragments as the coding ceruloplasmin gene sequence. Distribution of the nontranscribed beta-globin sequences in liver chromatin DNA fragments produced at different stages of nuclease digestion was not related to chromatin sensitivity to endogenous DNases. These sequences were distributed similarly in different DNA size classes and total DNA.

Animals↗

chromatin.

Explore the source record for details and available documents.

Acetates↗

[Spectral analysis of chromatin and its components in the vacuum ultraviolet region of the spectrum].

Electronic absorption spectra of thin films of chromatin and chromatin components in ultraviolet (140-280 nm) were investigated. The absorption coefficients mu (lambda) of chromatin, nucleosomes with and without histone H1, total histones (TH), DNA were compared. The spectra of nucleosomes and chromatin differ from summary spectra of DNA + TH. The lack of additivity of absorption coefficients at different wavelengths may be explained by different conformational changes of free DNA, TH and DNA, TH in nucleosomes and chromatin during the process of drying aqueous solutions for the preparations of thin films. The obtained mu (lambda) values are necessary for the estimation of the DNA and TH parts of absorption in chromatin and nucleosomes in the investigations of UV and VUV irradiation damages.

Animals↗

[DNAase I hypersensitivity of the 1,5'-flanking region of the tryptophan oxygenase gene in recombinant DNA and chromatin reconstituted with it].

We have examined DNase I hypersensitive sites in the 5'-flanking region of the rat tryptophan oxygenase gene in recombinant plasmids. We have found two DNase I-hypersensitive sites in regions that map between -350 to -210 and -180 to -90 bp from the cap site. Both hypersensitive sites are found in supercoiled plasmids but not in linear or relaxed DNA molecules. The position of the hypersensitive sites of the 5'-flanking sequence of the rat tryptophan oxygenase gene in recombinant plasmids correlate with the chromatin sites first determined by Becker et al. The hypersensitive regions in the recombinant plasmids include the DNase I-hypersensitive sites in chromatin but extend somewhat upstream and downstream from these. Computer analysis of the 5'-flanking DNA region of the gene suggests that the DNA fragment spanning nucleotides -231 to -207 forms a potential hairpin loop with 15 unpaired bases. Chromatin reconstitution with histones on supercoiled plasmids carrying the 5'-flanking region of the rat tryptophan oxygenase gene suppresses both in vitro DNase I-hypersensitive sites. We have also shown that the DNA region containing the supercoil-dependent DNase I-hypersensitive site in the position between -350 and -210 bp from the cap site may form nucleosomes after reconstitution with histones.

Animals↗