[Prognostic implications of GP3a glucoprotein gene PLA1/PLA2 allele in prostatic cancer: pilot results of the study].

We studied the role of integrins, primarily, the role of allele distribution of GP3a gene in development of prostatic cancer (PC) and assessment of its prognostic significance. From November 2003 to May 2004 we examined 32 patients with PC: 11 patients with local PC T1-2N0M0; 14 patients with locally advanced cancer T3N0M0 and 7 patients with invasive and/or metastatic cancer T3-4N10-1 or T3-4N0-1M1. The blood from all the patients we studied with PCR for alleles of GP3a gene, PSA.

Selection of E. coli strains for stable transformation with recombinant plasmids containing full-length genome of clinical HIV-1 isolates.

Strain chi6007 obtained from the parent E. coli strain chi5097 is a result of ptsH5 mutation, which allowed cells to grow without common components of the phosphoenolpyruvate-dependent phosphotransferase system. Segregants of strain chi6007 retaining the Pol+ gene responsible for inability to grow at 37 degrees C, but gaining rifampicin resistance (RifR) were used for cloning of cointegrate plasmids.

Alcohol dehydrogenase ADH2-1 and ADH2-2 allelic isoforms in the Russian population correlate with type of alcoholic disease.

The frequency ADH2-2 allele in the Moscow urban population and a correlation between the ADH2-2 allele, alcoholic dependence without cirrhosis, symptomatic alcoholic cirrhosis and status on hepatitis B and C infection have been studied. One hundred and twenty-three inhabitants of Moscow (50 healthy donors, 36 patients with alcoholic cirrhosis (subdivided into infected and uninfected by HBV and/or HCV) and 37 patients with alcoholic dependence) of a similar age/sex and drinking pattern have been analysed. The frequency of 41% for ADH2-2 allele is characteristic for an urban Moscow population.

Type D retrovirus specific sequences in lymphocytes of the children with Burkitt-type lymphoma and their parents.

Type D retroviruses cause immunodeficiency in monkey. Earlier we have revealed genetical and serological markers of type D retroviruses in children with Burkitt-type lymphoma. Using PCR/Southern blotting assay we have found sequences related to MPMV in PBMC's DNA from children with Burkitt-type lymphoma and from their parents.

Human spontaneous laryngeal carcinoma HEp-2 cells are chronically infected with SRV-1 virus, a variant of simian type D retrovirus.

A type D retrovirus chronically persisting in HEp-2 cells from human laryngeal carcinoma was analyzed by PCR and sequenced. This virus is most similar to SRV-1 and probably represents one of its subtypes.

Clinical symptoms of acute coronary insufficiency correlate with GP-IIIa genotype of integrin beta-subunit.

Studies of the clinical course of acute coronary insufficiency (progressive angina and myocardial infarction) in patients carrying a mutant allele of the Pl-A(II) gene encoding integrin GPIIIa beta-subunit revealed significant differences in the incidence of some clinical signs in comparison with patients homozygous for the normal Pl-A(I) allele.

Multiprotein complexes present at the MIF motifs flanking the promoter of the human c-myc gene.

The activated c-myc allele in Burkitt's lymphoma is associated with a clustering of somatic mutations within a discrete domain of intron I that define protein recognition sequences, designated as myc intron factors (MIF-1, MIF-2 and MIF-3). We have previously shown that MIF-1 binding activity consists of two polypeptides, myc intron binding polypeptide (MIBP1) and RFX1. In the present study we identified two polypeptides, p105 and p115, and showed that these proteins give rise to a DNA-protein complex at the MIF-2 as well as the adjacent MIF-1 site.

The role of phosphorylation of DNA-binding proteins in regulation of transcription of the human c-myc gene.

The goal of the present study was to identify transcriptional factors, to determine their specificity toward nucleotide sequences of binding sites, and to elucidate their regulatory effects on transcription of the human c-myc oncogene. Three novel phosphorylated proteins (transcriptional factors) participating in the regulation of transcription of the c-myc gene have been identified, and the following data have been obtained. A) The binding of the regulatory phosphoproteins activates in vitro transcription of the human c-myc gene.

The myc intron-binding polypeptide associates with RFX1 in vivo and binds to the major histocompatibility complex class II promoter region, to the hepatitis B virus enhancer, and to regulatory regions of several distinct viral genes.

We demonstrated that MIF-1, identified initially as a binding activity that associated with the intron I element of the c-myc gene, consists of two polypeptides, the myc intron-binding peptide (MIBP1) and the major histocompatibility class II promoter-binding protein, RFX1. Using a polyclonal antiserum directed against either oligonucleotide affinity-purified MIBP1 or a peptide derived from RFX1, we showed that MIBP1 and RFX1 are distinct molecules that associate in vivo and are both present in DNA-protein complexes at the c-myc (MIF-1) and major histocompatibility complex class II (RFX1) binding sites. We have also found that MIBP1 and RFX1 bind to a regulatory site (termed EP) required for enhancer activity of hepatitis B virus.

Enhancement of in vitro transcription of human c-myc correlates with the binding of phosphorylated protein factors from HeLa cell extract.

Hela cell extract overphosphorylated with endogenous protein kinases was used for in vitro transcription of human c-myc DNA. The activation of transcription initiated from P1, P1a and P2 cap-sites was observed. Three phosphoproteins of 70, 35 and 23 kDa are found, whose binding to DNA correlates with activation of c-myc transcription.

Cloning of a cDNA encoding a human protein which binds a sequence in the c-myc gene similar to the interferon-stimulated response element.

A human cDNA clone encoding a c-myc promoter-binding protein (IRLB) was selected by screening a human fibroblast lambda gt11 phage library with the hexamer oligodeoxyribonucleotide (oligo) 5'-GGCGGGAAAAAGAACGGA, corresponding to the protein-binding element of human c-myc similar to the interferon-stimulated response element (ISRE). The lambda gt11 phage clone, encoding a fusion protein which bound the probe oligo, was used to create an strain of Escherichia coli. The deduced amino-acid sequence of the cloned protein contains a putative alpha-helix which is expected to act as the DNA-binding domain.

[Transcription of antisense RNA for the human c-myc gene].

Antisense RNA transcription of human c-myc gene has been examined in HeLa, Burkitt lymphoma BL-60 t(8;22) cells, and diploid fibroblasts. By means of the primer extension technique two startpoints of antisense transcription have been detected and mapped with the first (untranslated) exon of the c-myc gene. Similarity between the antisense nucleotide sequence of the first c-myc intron and the SV40 DNA fragment containing the binding sites for transcriptions factors GT-I, GT-II, TC-I, and TC-II has been revealed by computer analysis.

Oligoadenylate and cyclic AMP: interrelation and mutual regulation.

The data obtained are in good agreement with the hypothesis that cAMP is involved in the control of 2-5A metabolism, including the mediation of the regulation of 2-5A by IFNs; 2-5A, in turn, affects the intracellular cAMP level. The general question originating from the data is that of a biochemical mechanism connecting the activation of the cAMP/2-5A system and the effect of depression of cell division. In my opinion, this universal effect is the result of the action of the known 2-5A-dependent mechanism, namely, RNase L (see review by Pestka et al.

Unexpected effect of an anti-human immunodeficiency virus intermolecular triplex-forming oligonucleotide in an in vitro transcription system due to RNase H-induced cleavage of the RNA transcript.

A 16-mer oligodeoxynucleotide (ODN) which specifically recognizes the polypurine tract (PPT) located upstream of the 3' long terminal repeat (LTR) of human immunodeficiency virus (HIV) proviral DNA via triplex formation is shown to have a dramatic effect on in vitro transcription from the HIV-LTR promoter. In the presence of HeLa cell extracts, a shorter RNA transcript is obtained in the presence of the 16-mer ODN. This truncated RNA lacks about 200 nucleotides from its 3' region.

Binding of proteins of HeLa S3 cell extract to oligonucleotides containing the consensus interferon-response sequence (IRS) and to the IRS-containing fragment of the human c-myc gene.

The human c-myc proto-oncogene was recently found to contain a regulatory sequence similar to the consensus interferon-response sequence (IRS) of interferon-activating genes. Binding of regulatory protein(s) to this sequence of cloned fragment of c-myc, lacking the main part of 5'-nontranscribing region, regulates in vitro transcription from I1/I2 initiation sites located in the first intron of the gene. Here, we have shown that HeLa S3 nuclear extract contains different protein factors, at least two, that bind preferentially to the IRS sequence of either the c-myc gene or the interferon-dependent 6-16 gene.

Human c-myc gene contains a regulatory site similar to consensus of interferon response sequence (IRS).

Expression of c-myc proto oncogene is regulated by multiple mechanisms. Here, we report that the consensus of the regulatory region of interferon-dependent genes, GGAAAN1-3 GAAA, was found after computer search in the 5'-terminal flank of human c-myc gene in position (-76:-67). In vitro transcription of c-myc gene fragments showed that the consensus region competes with oligonucleotide GGGAAAATGAAACT for binding to specific protein(s).

Expression of c-myc gene in human ovary carcinoma cells treated with vanadate.

The widely accepted hypothesis of vanadate action on cells postulates that this ion inhibits protein phosphatase(s) that dephosphorylates protein phosphotyrosine residues. This inhibition causes tyrosine hyperphosphorylation of cell proteins followed by changes in physiological action of phosphoproteins resulting in stimulation of cell proliferation, expression of protooncogenes, and transient cell transformation. We have found that treatment of human ovary carcinoma (CaOv) cells with vanadate causes the increase in total protein phosphorylation from 1.

[Effect of interferon on the cAMP system in cells].

The increase in cAMP concentration in CaOv cells affected by alpha-interferon has been found to have a two wave character with the maximums at 4 and 24 h after the effect. The waves are due to the increase in adenylate cyclase activity and to the decrease in the activity of cAMP phosphodiesterase. The described changes were characteristic of the native and partially purified interferon and depended on the concentration of interferon used (optimal effect at 1200 IU/ml-1).

[Directed synthesis of 2',5'-oligoadenylates with increased stability towards phosphodiesterases].

Phosphodiesterase stability of synthetic analogs of 2',5'-oligoadenylates, the mediators of antiviral and antiproliferative action of interferons was analysed. The analogs with a 3'-terminal acyclic nucleoside residue were prepared. These analogs were treated with NIH3T3 cell lysate, mice liver homogenate and snake venom phosphodiesterase.

A route to 2',5'-oligoadenylates with increased stability towards phosphodiesterases.

The rates of enzymatic hydrolysis of 2',5'-oligoadenylates and their synthetic analogs have been measured. These compounds were treated with either NIH 3T3 cell lysates, mouse liver homogenates or snake venom phosphodiesterase. All analogs with 3'-terminal acyclic nucleoside residues demonstrated greater stability compared with the natural compound adenylyl(2'-5')adenylyl(2'-5')adenosine.

[Induction of interferon-specific enzymes in cultures of cells sensitive and resistant to interferon].

Induction of 2'-5'-oligoadenylatesynthetase (2-5A synthetase) by interferons and theophylline by means of activation of cAMP-system in interferon susceptible and resistant cell lines were studied. In interferon resistant cell lines the basal activity of 2-5A synthetase exceeded the level of the same enzyme in interferon susceptible cell lines. Activity of 2-5A synthetase is increased in interferon susceptible cell lines by interferon treatment, but the activity of the enzyme is not altered in interferon resistant cell lines.

[Effect of modulation of the production of interferon by L-929 cells treated with theophylline].

The specific inhibitor of cAMP phosphodiesterase theophylline has been shown to evoke in L929 cells 2.3-fold induction of 2-5A-synthetase activity and 3.5-fold superinduction of the same enzyme activity while acting in combination with actinomycin D.