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Vol 486, No 1 (2019)

Biochemistry, Biophysics, and Molecular Biology

Cytokinins and Abscisic Acid Regulate the Expression of the Genes for Plastid Transcription Apparatus during Heat Shock

Andreeva A.A., Bychkov I.A., Danilova M.N., Kudryakova N.V., Kusnetsov V.V.

Abstract

The treatment of Arabidopsis thaliana plants with exogenous cytokinin (CK) followed by heat shock (HS) activated the expression of the genes for the plastid transcription machinery but adversely affected the plant viability. Abscisic acid (ABA), conversely, promoted maintaining the resistance to HS and had differentially affected different components of the plastid transcriptional complex. This hormone suppressed the accumulation of transcripts of PEP genes and the genes encoding PAP proteins, which are involved in DNA–RNA metabolism. However, it had no effect or activated the expression of NEP genes and PAP genes, which are involved in the redox regulation, as well as the genes encoding the stress-inducible trans-factor (SIG5) and the plastid transcription Ser/Thr protein kinase (cpCK2). Thus, for the adaptation of plants to elevated temperatures, both increase and decrease in the expression of the genes for the plastid transcriptional machinery with the involvement of various regulatory systems, including phytohormones, are equally significant.

Doklady Biochemistry and Biophysics. 2019;486(1):163-167
pages 163-167 views

Synuclein Deficiency Decreases the Efficiency of Dopamine Uptake by Synaptic Vesicles

Ninkina N.N., Tarasova T.V., Chaprov K.D., Goloborshcheva V.V., Bachurin S.O., Buchman V.L.

Abstract

In the present study, we analyzed the uptake of radiolabeled dopamine by intact synaptosomes and purified synaptic vesicles isolated from the dorsal striatum of mice with constitutive inactivation of all three synuclein-coding genes and wild-type mice. Synuclein deficiency substantially compromised the uptake of this neurotransmitter by synaptic vesicles but had no effect on synaptosomal dopamine uptake.

Doklady Biochemistry and Biophysics. 2019;486(1):168-170
pages 168-170 views

Estimation of Metabolism of Catecholamines in Peripheral Organs As an Indicator of Their Desympathization under the Influence of Neurotoxins

Safandeev V.V., Kolacheva A.A., Ugrumov M.V.

Abstract

Under conditions of the experimental model of Parkinson’s disease at the preclinical (early) and clinical stage—injection of mice with the neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) for 5 and 10 weeks—the toxic effects of MPTP was evaluated (the content of catecholamines and their metabolites in the heart, which receives the most extensive sympathetic innervation, was determined). The obtained data indicated the beginning of desympathization of the heart at the preclinical stage of PD and its progression at the clinical stage of the disease.

Doklady Biochemistry and Biophysics. 2019;486(1):171-174
pages 171-174 views

Studying Interactions between the Mod(mdg4)-67.2 Protein and Other Mod(mdg4) Isoforms in the Embryonic Cells of Drosophila melanogaster

Kostyuchenko M.V., Melnikova L.S., Georgiev A.P., Golovnin A.K.

Abstract

It is found that, in embryonic D. melanogaster cells, Mod(mdg4) protein isoforms can interact with each other through BTB domains. However, this nonspecific interaction is destroyed as a result of recruitment of protein complexes to the chromatin sites.

Doklady Biochemistry and Biophysics. 2019;486(1):175-180
pages 175-180 views

Thapsigargin, Inhibitor of Sarco-Endoplasmic Ca2+-ATPase, Effectively Suppresses the Expression of S100A4 Protein in Human Breast Cancer Cell Line

Kotnova A.P., Lyanova B.M., Dukhanina E.A., Portseva T.N., Ilyin Y.V., Georgieva S.G., Stepchenko A.G., Pankratova E.V.

Abstract

Thapsigargin (SERCA ATPase inhibitor) inhibited the S100A4 metastatic marker expression in MDA-MB231 breast cancer cells. We found that S100A4 gene transcription is regulated by Ca2+ signaling pathways. We found that the synthesis of S100A4 mRNA and S100A4 protein in MDA-MB231 cells was effectively suppressed by thapsigargin at a concentration of 0.4–4 μM with retaining cell viability. We assume that the change in the gene transcription in response to disturbance of Ca2+ homeostasis is directly involved in the remodeling of Ca2+ signaling pathways.

Doklady Biochemistry and Biophysics. 2019;486(1):181-183
pages 181-183 views

Bicistronic Construct for Optogenetic Prosthesis of Ganglion Cell Receptive Field of Degenerative Retina

Petrovskaya L.E., Roshchin M.V., Smirnova G.R., Kolotova D.E., Balaban P.M., Ostrovsky M.A., Malyshev A.Y.

Abstract

To perform optogenetic prosthetics of the retinal ganglion cell receptive field, a bicistronic genetic construct carrying the genes encoding the excitatory (channelrhodopsin-2) and inhibitory (Guillardia theta anion channelrhodopsin GtACR2) rhodopsins was created. A characteristic feature of this construct was the combination of these two genes with a mutant IRES insertion between them, which ensures the exact ratio of expression levels of the first and second genes in each transfected cell. Illumination of the central part of the neuron with light with a wavelength of 470 nm induced the action potential generation in the cell. Stimulation of the peripheral neuronal region with light induced the inhibition of action potential generation. Thus, using optogenetics methods, we simulated the ON–OFF interaction in the retinal ganglion cell receptive field. Theoretically, this construct can be used for optogenetic prosthetics of degenerative retina in the case of its delivery to the ganglion cells with lentiviral vectors.

Doklady Biochemistry and Biophysics. 2019;486(1):184-186
pages 184-186 views

Study of dCTCF Insulator Activity in Drosophilamelanogaster Model Systems

Postika N.E., Ivlieva T.A., Georgiev P.G., Kyrchanova O.V.

Abstract

Using transgenic Drosophila model systems, we showed that four binding sites for the architectural protein dCTCF per se cannot form an effective insulator that blocks enhancers and protects against the Polycomb-dependent repression. These results suggest that, in the known Drosophila insulators, the dCTCF protein functions in cooperation with other architectural proteins.

Doklady Biochemistry and Biophysics. 2019;486(1):187-191
pages 187-191 views

The CYP443C1 (CYP74 clan) Cytochrome of Sea Anemone Nematostella vectensis–the First Metazoan Enzyme Possessing Hydroperoxide Lyase/Epoxyalcohol Synthase Activity

Gorina S.S., Toporkova Y.Y., Mukhtarova L.S., Grechkin A.N.

Abstract

A novel CYP74 clan gene CYP443С1 of the starlet sea anemone (Nematostella vectensis, Cnidaria) has been cloned, and the properties of the corresponding recombinant protein have been studied. Depending on the substrate, CYP443С1 exhibited double function hydroperoxide lyase/epoxyalcohol synthase activity.

Doklady Biochemistry and Biophysics. 2019;486(1):192-196
pages 192-196 views

Peroxide-Induced Oxidative Modification of Hemoglobin

Vasilyeva A.D., Yurina L.V., Bugrova A.E., Indeykina M.I., Azarova D.Y., Bychkova A.V., Akzhigitova K.I., Kononikhin A.S., Nikolaev E.N., Rosenfeld M.A.

Abstract

The oxidative modification of human hemoglobin (Hb) treated with hydrogen peroxide was investigated. Using the mass spectrometry method, the oxidized amino acid residues of the hemoglobin molecule were detected: αTrp14, αTyr24, αArg31, αMet32, αTyr42, αHis45, αHis72, αMet76, αPro77, αLys90, αCys104, αTyr140, βHis2, βTrp15, βTrp37, βMet55, βCys93, βCys112, βTyr130, βLys144, and βHis146. The antioxidant potential of the Hb molecule in the intracellular space and in the blood plasma is discussed.

Doklady Biochemistry and Biophysics. 2019;486(1):197-200
pages 197-200 views

NSP Protein Encoded in Negative NS RNA Strand of Influenza A Virus Induces Cellular Immune Response in Infected Animals

Zhirnov O.P., Isaeva E.I.

Abstract

Infection of mice with influenza A viruses led to the formation of clones of lymphocytes that specifically recognizes viral domains in the central zone of the NSP protein (amino acid positions 83–119). Computer analysis of the primary structure of the NSP protein showed the presence of T-cell epitopes in the central part of the NSP molecule. The findings indicate that the viral NSP gene is expressed in the infected animals and verify the concept of the bipolar strategy (ambisense strategy) of the influenza A virus genome.

Doklady Biochemistry and Biophysics. 2019;486(1):201-205
pages 201-205 views

Bacterial Film Disintegration with Electrochemically Reduced Water

Pogorelov A.G., Kuznetsov A.L., Panait A.I., Pogorelova M.A., Suvorov O.A., Ivanitskii G.R.

Abstract

The aim of this work was to study the fine structure of bacterial films grown on the inner tube surface of a flow reactor. Using the scanning electron microscopy (SEM) approaches, the detailed biofilm relief was visualized. The action of electrochemically reduced water (ERW) on the biofilm ultrastructure generated by the plankton form of E. coli and/or lacto bacteria was investigated. The treatment with an ERW solution destroyed the biofilm organic polymer matrix and bacterial cells embedded in the matrix.

Doklady Biochemistry and Biophysics. 2019;486(1):206-208
pages 206-208 views

Luciferin–Luciferase System of Marine Polychaete Chaetopterus variopedatus

Purtov K.V., Petushkov V.N., Rodionova N.S., Pakhomova V.G., Myasnyanko I.N., Myshkina N.M., Tsarkova A.S., Gitelson J.I.

Abstract

This paper presents the preliminary results of the separation of the Chaetopterus variopedatus bioluminescent system into luciferin and luciferase and a brief description of some of their properties.

Doklady Biochemistry and Biophysics. 2019;486(1):209-212
pages 209-212 views

NAD(P)H:FMN‑Oxidoreductase Functioning Under Macromolecular Crowding: In Vitro Modeling

Govorun A.E., Esimbekova E.N., Kratasyuk V.A.

Abstract

The functioning of NAD(P)H:FMN‑oxidoreductase (Red) from Vibrio fischeri under conditions of macromolecular crowding (MMC) simulated in vitro by adding biopolymers (starch and gelatin) was studied. The dissociation rate constants and the activation energies of dissociation of Red to the subunits were calculated, and the process of denaturation of Red was analyzed. It is shown that the functioning of Red both under conditions of MMC and in diluted solutions is the same. This result refutes the common belief that the native conformation of enzymes in vivo is stabilized due to MMC as compared to the in vitro conditions.

Doklady Biochemistry and Biophysics. 2019;486(1):213-215
pages 213-215 views

Bacteriochlorophyll Interaction with Singlet Oxygen in Membranes of Purple Photosynthetic Bacteria: Does the Protective Function of Carotenoids Exist?

Makhneva Z.K., Ashikhmin A.A., Bolshakov M.A., Moskalenko A.A.

Abstract

The direct action of singlet oxygen on the bacteriochlorophyll (BChl) of light-harvesting complexes in the membranes of four species of purple non-sulfur and sulfur photosynthesizing bacteria with and without carotenoids was studied. It was found that BChl in carotenoidless samples is generally more resistant to the action of singlet oxygen compared to the control. It is assumed that carotenoids are not required to protect BChl of bacterial light-harvesting complexes from singlet oxygen, and in the classic work by Griffith et al. [1] the apoptosis process in carotenoidless mutant cells, which involves the destruction of complexes, the appearance of monomeric BChl, and the generation of singlet oxygen caused by BChl, followed by BChl oxidation, was mistakenly attributed to the protective function of carotenoids.

Doklady Biochemistry and Biophysics. 2019;486(1):216-219
pages 216-219 views

Molecular Dynamics Modeling of the Interaction of Cationic Fluorescent Lipid Peroxidation-Sensitive Probes with the Mitochondrial Membrane

Nesterenko A.M., Kholina E.G., Lyamzaev K.G., Mulkidjanian A.Y., Chernyak B.V.

Abstract

Cardiolipin (CL) plays a central role in lipid peroxidation (LPO) of the mitochondrial inner membrane due to higher content of unsaturated fatty acids in CL in comparison with the other phospholipids. CL oxidation plays an important role in the regulation of various intracellular signaling pathways and its excessive oxidation contributes to the development of various pathologies and, possibly, participates in the aging process. Mitochondria-targeted antioxidants containing triphenylphosphonium (TPP+) effectively protect CL from oxidation. It is assumed that fluorescent probes on the basis of the C11-BODIPY fluorophore sensitive to LPO and containing TPP+ can selectively register CL oxidation. To test this possibility, we carried out a molecular dynamic simulation of such probes in a model mitochondrial membrane. It is shown that the probes are located in the membrane at the same depth as the unsaturated bonds in CL molecules sensitive to oxidation. Increasing the length of the linker that binds the fluorophore and TPP+ residue ha little effect on the position of the probe in the membrane. This indicates the possibility of modifying the linker to increase the selectivity of the probes to CL.

Doklady Biochemistry and Biophysics. 2019;486(1):220-223
pages 220-223 views

Functional Comparison of Short and Long Isoforms of the TRF2 Protein in Drosophila melanogaster

Osadchiy I.S., Georgiev P.G., Maksimenko O.G.

Abstract

TRF2 protein (TBP-related factor 2) can substitute for TBP forming alternative transcription initiation complexes on TATA-less promoters, including the promoters of histone H1 and piRNA clusters required for transposon repression. The Drosophilatrf2 gene codes for two isoforms: a “short” and a “long” one, in which the same short TRF2 sequence is preceded by a long N-terminal domain. Here, we demonstrated that the long TFR2 isoform has a greater functional activity than the short isoform by expressing each of them at a reduced rate under the endogenous promoters. Expression of the long isoform alone affects neither the flies’ viability nor the sex ratio. Expression of the short isoform alone leads to the phenotype described for the trf2 gene insufficiency and derepression of transposable elements, that is, decreased viability, disturbance of homologous chromosome pairing and segregation, and apparent female-biased sex ratio.

Doklady Biochemistry and Biophysics. 2019;486(1):224-228
pages 224-228 views

Lipid Fatty Acids from the Pericarp of Cydonia oblonga Mill. and Mespilus germanica L. are Involved in Plant Adaptation to Altitudinal Zonality

Ivanova T.V., Voronkov A.S., Kuznetsova E.I., Kumachova T.K., Zhirov V.K., Tsydendambaev V.D.

Abstract

The composition of fatty acids of total lipids of the outer and parenchymal parts of the pericarp in Cydonia oblonga Mill. and Mespilus germanica L. (Maloideae, Rosaceae), growing in the Northern Caucasian mountains at altitudes of 300, 500, 700, and 1200 m above sea level in various natural zones from experimental sites, was studied for the first time. It is established that the altitude of plant growth is largely correlated with the changes in the FA composition of the outer, but not the parenchymal, part of the pericarp. The nature of this variability suggests that the adaptation of plants to the conditions of significant temperature differences in the mountains is associated with the regulation of cell membrane fluidity, based on the interaction of opposite processes of synthesis of polyunsaturated and very-long-chain fatty acids.

Doklady Biochemistry and Biophysics. 2019;486(1):229-233
pages 229-233 views

The Spatial Metabolic Heterogeneity of Solid Type of Enrlich Carcinoma

Inzhevatkin E.V., Savchenko A.A.

Abstract

In this work, we investigated the NAD(P)-dependent dehydrogenase activity in different fields of the solid form of Ehrlich carcinoma. It was shown that there is a metabolic distinction between different fields of the solid tumor. In this way, there is a significant difference between the ascites and solid type of Ehrlich carcinoma. In the central area of the tested sample of tumor tissue, there is a high intensity of energetic metabolism and biosynthetic processes as opposed to the peripheral areas. Previously, we proposed the concept of nonspecific metabolic reaction of the cell in organism upon the influence of negative factors. In accordance with this concept, our results indicate the development of adaptation in the tumor cells in the central area of the tested sample.

Doklady Biochemistry and Biophysics. 2019;486(1):234-237
pages 234-237 views

Anticancer Activity of Dinitrosyl Iron Complex (NO Donor) on the Multiple Myeloma Cells

Akentieva N.P., Sanina N.A., Prichodchenko T.R., Gizatullin A.R., Shkondina N.I., Shushanov S.S., Stupina T.S., Aldoshin S.M.

Abstract

The results of the study of the effect of a mononuclear dinitrosyl iron complex (DNIC7) with functional sulfur-containing ligands (NO donors) on the viability of multiple myeloma cells are presented. It was shown that DNIC7 decreased cell viability and inhibited the proliferation of multiple myeloma cells, i.e., exhibits cytotoxic properties. Fluorescent analysis showed that the DNIC7 compound decreases the level of intracellular glutathione and increases the level of reactive oxygen species in multiple myeloma cells. It is assumed that DNIC7 has a therapeutic potential for the treatment of cancer.

Doklady Biochemistry and Biophysics. 2019;486(1):238-242
pages 238-242 views

The Role of the Brain in the Regulation of Peripheral Noradrenaline-producing Organs in Rats During Morphogenesis

Murtazina A.R., Nikishina Y.O., Dil’mukhametova L.K., Sapronova A.Y., Ugrumov M.V.

Abstract

This work represents one part of our research project, in which we attempted to prove that a humoral regulation between noradrenaline-producing organs exist in the perinatal period. In this study, we used a rat model that allowed blocking the synthesis of noradrenalin in the brain and evaluated gene expression and protein levels of noradrenaline key synthesis enzymes such as tyrosine hydroxylase (TH) and dopamine beta-hydroxylase (DBH) in peripheral noradrenaline-producing organs. As a result, we showed an increased gene expression of TH and DBH in adrenal glands. These data indicate that, if neonatal rat brain lacks the ability to produce noradrenaline, then the synthesis of noradrenaline in adrenal glands increased as a compensatory process, so that the concentration levels in blood are maintained at normal levels. This indicates that there is a humoral regulation between brain and adrenal glands, which is not fully understood yet.

Doklady Biochemistry and Biophysics. 2019;486(1):243-246
pages 243-246 views