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Biomedical subjects

S L Kiselev

Publications and source records attributed to S L Kiselev.

At least 37 records · Page 2Linked to original sources

[tag7 Gene and gene therapy of cancer].

Modern gene therapeutic approaches towards the treatment of malignant neoplasms are discussed. Specifically, results of the first experiments on the development of vaccines on the basis of tumor cells modified with various cytokines and activation molecules are described. We compare effectiveness of different cytokines used for modification of tumor cells are made and consider possible mechanisms of vaccine action. The reasons for impossibility of the development of effective endogenous antitumor immunity are examined. The results of comparative analysis of the Tag7 protein antitumor activity are presented. This evidence clearly shows that Tag7 is a promising agent for anticancer therapy.

Animals↗

[Cloning and study of novel mammalian genes with structural homology with phage lysozyme].

The mouse tag7 gene was cloned and characterized in our previous works. This work is devoted to identifying and cloning its homologs. The human homolog of the tag7 gene was cloned. Data of analysis of the primary structure of the human tag7 and results from the study of the production of the corresponding protein in human organs and tissues indicate that this gene plays a major role in the mammalian immune system. The mouse and human tagL gene carrying an extended region of structural homology with the tag7 gene was detected by computer analysis and was subsequently cloned. Sequence analysis suggested the possible membrane localization of the gene, whereas the specific expression pointed to the role of this gene in the immune system. Both genes, tag7 and tagL, were localized in the human chromosome 19.

Amino Acid Sequence↗

Resistance to tumor necrosis factor induced apoptosis in vitro correlates with high metastatic capacity of cells in vivo.

TNF is one of the cytokines secreted by the cells of the immune system. Our data demonstrate that those cell lines lacking capability to form metastatic tumors in vivo are susceptible to TNF induced apoptosis in vitro. However, cell lines with high metastatic potential are resistant to TNF in vitro. Furthermore, the same cell lines were resistant to cytolytic action of other cytotoxic proteins secreted by LAK cells. Our data showed that TNF resistance in vitro correlates with the increased level of transcription factor NF-kappaB. This finding may provide a tool to improve current protocols of immunotherapy and insights to how tumor cells are or are not killed by LAK cells.

Animals↗

Molecular cloning and characterization of the mouse tag7 gene encoding a novel cytokine.

Cloning of the mouse tag7 gene encoding a novel cytokine is described. The Tag7 protein consists of 182 amino acids. Genomic organization of the tag7 gene and its promoter region remind those of the genes of the tumor necrosis factor locus, although the tag7 gene is not linked to this locus. The gene is located on chromosome 7 at the area that corresponds to band 7A3, which has genetic linkage with lupus-like disease in mouse models. tag7 transcription is essential for lymphoid organs. It is also detected in certain areas of lungs, brain, and intestine and in some tumors. Tag7 protein is detectable in both cell-associated and soluble forms. The soluble form of Tag7 triggers apoptosis in mouse L929 cells in vitro and does not involve NF-kappaB activation. The relationship between Tag7 and tumor necrosis factor family of ligands is discussed.

Amino Acid Sequence↗

A minisatellite "core" element constitutes a novel, chromatin-specific activator of mts1 gene transcription.

Expression of the mts1 gene is often associated with malignant transformation of tumor cells. Transcription of the gene is controlled by a number of positive and negative regulatory elements, all of them being localized in the first intron (+38 to +1215) of the mts1 gene. Through analysis of the distribution of DNase I hypersensitive sites in the first intron of the gene we revealed a structurally conserved region that consisted of a non-canonical NFkB binding site and a minisatellite "core" element. Deletion of the minisatellite core DNA in the context of the first intron had no effect on its regulatory capacity when assayed in transient transfections, while a fivefold decrease was observed in a pool of stably transfected cells. The minisatellite core sequence CTGGGCAGGCAG is involved in DNA-protein interactions in vivo, and is similar to a binding site for the previously identified minisatellite DNA sequence binding protein (Msbp-1). The core DNA interacted in vitro with a protein that had an apparent molecular mass of 40 kDa. These data indicate that the minisatellite DNA represents the novel, chromatin-specific element in the mts1 complex enhancer.

Animals↗

[Cloning of the tag7 gene expressed in metastatic mouse tumors].

Gene expression was compared in a metastatic (VMR-Liv) neoplastic cell line and a related nonmetastatic (VMR-O) neoplastic cell line by means of the differential display method. A fragment of cDNA corresponding to the tag7 gene, differentially expressed in the metastatic cell line, was isolated. The full-length tag7 cDNA was cloned and its nucleotide sequence was determined. No homology between the tag7 gene and known sequences was revealed. tag7 gene transcription was studied in some tumors, cell lines, and normal mouse organs.

Amino Acid Sequence↗

Differential splicing creates a diversity of transcripts from a neurospecific developmentally regulated gene encoding a protein with new zinc-finger motifs.

We have cloned a novel neurospecific gene, named neuro-d4, by differential screening a rat cerebral cortex cDNA library. Northern blot hybridization showed that neuro-d4 expression is restricted to neuronal tissues both in newborn and adult animals. The level of neuro-d4 mRNA in the rat central nervous system is high during the later stages of embryonic development and gradually decreases during the postnatal period. In situ hybridization suggests that the gene transcripts are localized in neuronal cell bodies. Nucleotide sequences of overlapped cDNA clones and all 12 exons in genomic clone were determined. The deduced protein has consensus sequences for a nuclear localization signal, a Krüppel-type zinc-finger and a new type of cysteine/histidine-rich motif resembling zinc-fingers. Several differential splicing variants were found, each of which influences the structure of the encoded protein.

Amino Acid Sequence↗

A novel transposition system in Drosophila melanogaster depending on the Stalker mobile genetic element.

Crosses between the Drosophila melanogaster y2sc1waG strain or some of its derivatives and the FM4 strain yielded insertional mutagenesis with a frequency of 10(-3)-10(-4). The system differs in several respects from the known cases of hybrid dysgenesis: (i) it does not depend on the direction of a cross; (ii) destabilization continues for a long time after initial crosses; (iii) mutations may occur at different stages of development. The mutation in the yellow locus has been cloned and found to depend on insertion into the coding region of the gene of a novel mobile genetic element designated as Stalker. The sequencing of Stalker termini reveals 405 bp direct repeats (LTRs) and a target 3 bp duplication, as well as some other sequences typical of retrovirus-like retrotransposons. The number of Stalker copies per genome and chromosomal localization vary among D. melanogaster strains. Before crosses, the location of Stalker on chromosomes is fairly stable in a particular strain but thereafter numerous changes in Stalker distribution take place. Most novel substrains are internally heterogenous which is indicative of the continuing Stalker transposition. Other mobile elements tested do not move. Possibly, only Stalker is mobilized in the system. Many known and novel mutations have been obtained. Comparison of their genetic localization with Stalker distribution suggests that the majority of them have been induced by the Stalker insertion.

Animals↗

[Transpositional bursts and chromosome rearrangements in unstable lines of Drosophila].

The phenomenon of transpositional bursts-massive simultaneous transpositions of mobile elements belonging to different structural classes and accompanied by multiple mutagenesis were earlier described. Although the mechanisms of this phenomenon are still unclear, it is obvious now that it embraces total genome and includes not only transpositions of different mobile elements but also recombination processes--homologous recombination for LTR's and gene conversion. It is shown in this work that transpositional bursts may be accompanied by appearance of grass chromosomal rearrangements. The chain of closely related mutations which is characterized, as well as pedigrees described earlier, by coordinated mutational transitions and multiple transpositions of mdg1, mdg2 and retrotransposon jokey was analyzed. Spontaneous appearance of mutations in the loci yellow, white (deficiency for 462 kb) and cut (insertion of mdg4, together with jokey) correlates with appearance of inversion In(I), 14A-20B, and the reversions of these mutations to the wild type (y+w+ct+) or to other alleles (ctMR2--insertion of mdg4 without jokey) are accompanied by reversions of inversion. The relationship of all lines analyzed in this work as well as the lines from other pedigrees was proved using analysis of polymorphic restriction sites at the scute and cut loci (5 probes were used). All "y w ctpN"--type mutants are shown to have insertion of about 7 kb at the scute locus which causes no alteration of phenotype. This once again proves multiple and coordinated character of changes taking place during hybrid dysgenesis.

Animals↗