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Barygina V.V.,Moscow State University | Veiko V.P.,Research Institute of Genetics and Selection of Industrial Microorganisms | Zatsepin O.V.,RAS Shemyakin Ovchinnikov Institute of Bioorganic Chemistry
Biochemistry (Moscow) | Year: 2010

Fibrillarin is an evolutionarily-conserved and obligatory protein component of eukaryotic cell nucleoli involved in pre-rRNA processing and methylation. In vertebrates the fibrillarin molecule contains two cysteine residues (Cys99 and Cys268) whose sulfhydryl groups are able to establish intramolecular -S-S- bridges. However, the functional state of fibrillarin with reduced or oxidized thiol groups is still practically unstudied. Besides, there are no data in the literature concerning existence of the -S-S- fibrillarin form in human cells. To answer these questions, we used plasmids encoding native human fibrillarin and its mutant form devoid of cysteine residues (fibrillarinC99/268S) fused with EGFP for temporary transfection of HeLa cells. The mobile fraction localizing the enzymatically active protein molecules and the fluorescence half-recovery time characterizing the rate of enzymatic reactions were determined by the FRAP technique using a confocal laser scanning microscope. Measurements were carried out at 37 and 27°C. The results show that the fibrillarin pool in HeLa cells includes two protein forms, with reduced SH groups and with oxidized SH groups forming intramolecular -S-S- bridges between Cys99 and Cys268. However, the absence of Cys99 and Cys268 has no effect on intracellular localization of fibrillarin and its main dynamic parameters. The human fibrillarin form without disulfide bridges is included into the mobile protein fraction and is consistent with its functionally active state. © 2010 Pleiades Publishing, Ltd. Source


Vorob'eva L.I.,Moscow State University | Khodzhaev E.Y.,Moscow State University | Vustin M.M.,Research Institute of Genetics and Selection of Industrial Microorganisms
Applied Biochemistry and Microbiology | Year: 2011

Antistress effect of extracellular peptides on UV-irradiated yeast of different phylogenetic groups was studied. Yeast from different ecotopes and taxonomic groups exposed to UV radiation of a lethal intensity showed a protective effect and reactivating effect with participation of extracellular peptides. The highest protective activity was found in peptide reactivation factors (RFs) of bakery yeast-Saccharomyces cerevisiae, Kluyveromyces fragilis, and Candida utilis; the highest reactivating activity was exhibited by factors of the above-mentioned cultures and Debariomyces hansenii. Cross-protective and reactivating effects of RFs of yeast belonging to different taxonomic groups were demonstrated. Cross-protection increased two to three times after preexposure of reactivation factors to UV light (activation) in contrast to their reactivating effect. © 2011 Pleiades Publishing, Ltd. Source


Rykov S.V.,Research Institute of Genetics and Selection of Industrial Microorganisms | Khodyrev D.S.,Research Institute of Genetics and Selection of Industrial Microorganisms | Pronina I.V.,Russian Academy of Medical Sciences | Kazubskaya T.P.,Russian Academy of Medical Sciences | And 2 more authors.
Russian Journal of Genetics | Year: 2013

MicroRNAs play an important role in the regulation of expression of many genes and are involved in carcinogenesis. The regulation of miRNA gene expression can involve the methylation of promoter CpG islands. In this work, the methylation of six miRNA genes (mir-107, mir-125b-1, mir-130b, mir-137, mir-375, and mir-1258) in non-small-cell lung cancer (NSCLC) was studied for the first time by methylation-specific PCR using a representative set of specimens (39 cases). Four new genes (mir-125b-1, mir-137, mir-375, and mir-1258) methylated in primary NSCLC tumors were identified with frequencies of 56, 31, 56, and 36%, respectively. The frequencies of miRNA promoter methylation in DNA of tumors and histologically normal tissues differed significantly (P ≤ 0.05 by Fisher's test). In lung tissues of 20 donors without a history of cancer, these genes were only methylated in a few cases. It was also shown that the previously unstudied promoter CpG islands of mir-107 and mir-130b were not methylated in NSCLC. The frequencies of mir-125b-1 and mir-137 methylation were shown for the first time to correlate with NSCLC progression (clinical stage and metastasis). © 2013 Pleiades Publishing, Ltd. Source


Orlova N.V.,Research Institute of Genetics and Selection of Industrial Microorganisms | Hamitov R.A.,Research Institute of Genetics and Selection of Industrial Microorganisms
Russian Journal of Biopharmaceuticals | Year: 2013

A new method for quantitative determination of erythropoietin and darbepoietin in culture medium using affinity chromatography with fluorimetric detection is described. The method was validated in terms of specificity, linearity, accuracy, repeatability, intermediate precision, quantitation limit and range. Method could be applied to qualitative assessment culture medium in terms of quantification of the active component. Source


Orlova N.V.,Research Institute of Genetics and Selection of Industrial Microorganisms | Gavrilova N.A.,Research Institute of Genetics and Selection of Industrial Microorganisms | Khamitov R.A.,Research Institute of Genetics and Selection of Industrial Microorganisms
Russian Journal of Biopharmaceuticals | Year: 2014

The new method for quantitative determination of erythropoietin and darbepoietin in culture medium using reverse phase chromatography with UV-detection is described. The method was validated in terms of specificity, linearity, accuracy, repeatability, intermediate precision, quantitation limit and range. Method could be applied to qualitative assessment culture medium in terms of quantification of the active component. © 2014 Folium Ltd. All rights reserved. Source

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