ЕС

Е. С. Клименко

Lomonosov Moscow State University

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Yu. M. Konstantinov

Siberian Institute of Plant Physiology and Biochemistry

Maria Breygina

Lomonosov Moscow State University

M. V. Koulintchenko

Siberian Institute of Plant Physiology and Biochemistry

ВИ

В. И. Тарасенко

Siberian Institute of Plant Physiology and Biochemistry

ВН

В. Н. Шмаков

Siberian Institute of Plant Physiology and Biochemistry

Anna Kostareva

Federal Almazov North-West Medical Research Centre

I. Yu. Subota

Siberian Institute of Plant Physiology and Biochemistry

Alexandra Podolyan

Lomonosov Moscow State University

Oksana Luneva

Lomonosov Moscow State University

André Dietrich

Institut de Biologie Moléculaire des Plantes

Cooperative Institutions

Russian Academy of Sciences

Federal Almazov North-West Medical Research Centre

Siberian Branch of the Russian Academy of Sciences

Siberian Institute of Plant Physiology and Biochemistry

Ministry of Health of the Russian Federation

Lomonosov Moscow State University

St Petersburg University

Centre National de la Recherche Scientifique

Institut de Biologie Moléculaire des Plantes

Institute of Chemical Biology and Fundamental Medicine

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Studying of import Mechanisms for different length and structure DNA into plant mitochondria

Journal of stress physiology & biochemistry (2014)

Е. С. Клименко M. V. Koulintchenko П. А. Гребнев André Dietrich Yu. M. Konstantinov

We investigated import of DNA of various length (109 bp, 269 bp, 717 bp, 1540 bp, 2732 bp, 9000 bp and 11600 bp) into the isolated mitochondria from potato tubers ( Solanum tuberosum ) and turnip root crops ( Brassica rapa ). The results imply existence of multiple pathways of DNA transfer into plant mitochondria. It was established that the transport of short length DNA (100-300 bp) is apparently carried out with the participation of several protein carriers which nature is currently unknown. We identified factors that influence the process of DNA transfer into mitochondria. These factors include the length of nucleic acid molecule and the presence of terminal inverted repeats that are specific to linear mitochondrial plasmids of plants.

Brassica rapa

Solanum tuberosum

Inverted repeat

Nuclear DNA

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Generation of an Induced Pluripotent Stem Cell Line ICGi046-A of a Patient Carrying Pathogenic p.Ser259Thr Variant in RAF1 Associated with RASopathy

Russian Journal of Developmental Biology (2023)

Elena Vladimirovna Dementyeva Е. С. Клименко J. M. Minina Suren M. Zakian Anna Kostareva

Developmental Biology

Line (geometry)

10.1134/s1062360423070032

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STUDYING OF ROLE OF THE INTERACTION BETWEEN MITOCHONDRIA MEMBRANES AND ENDOPLASMATIC RETICULUM IN DNA IMPORT

Е. С. Клименко В. Н. Шмаков Т.А. Болотова I. Yu. Subota В. И. Тарасенко

Reticulum

10.31255/978-5-94797-318-1-49-51

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Tonoplast Rafts Stimulate DNA Import into the Mitochondria of Potato Tubers (Solanum tuberosum)

Biochemistry (Moscow) Supplement Series A Membrane and Cell Biology (2020)

Е. С. Клименко И. С. Нестеркина Н. В. Озолина В. В. Гурина M. V. Koulintchenko

Solanum tuberosum

Raft

10.1134/s1990747820040066

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Citations (1)

Specificity of DNA import into isolated mitochondria from plants and mammals

Biopolymers and Cell (2014)

M. V. Koulintchenko Noha Ibrahim Е. С. Клименко Yu. M. Konstantinov F. Weber-Lotfi

Aim. Investigation of different features of DNA import into plant and human mitochondria, for a better understanding of mitochondrial genetics and generation of biotechnological tools.Methods.DNA up-take experiments with isolated plant mitochondria, using as substrates various sequences associated or not with the specific terminal inverted repeats (TIRs) present at each end of the plant mitochondrial linear plasmids.Results.It was established that the DNA import efficiency has a non-linear dependence on DNA size.It was shown that import into plant mitochondria of DNA molecules of «medium» sizes, i. e. between 4 and 7 kb, barely has any sequence specificity: neither TIRs from the 11.6 kb Brassica plasmid, nor TIRs from the Zea mays S-plasmids influenced DNA import into Solanum tuberosum mitochondria.Conclusions.The data obtained support the hypothesis about species-specific import mechanism operating under the mitochondrial linear plasmids transfer into plant mitochondria.

10.7124/bc.000880

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The Balance between Different ROS on Tobacco Stigma during Flowering and Its Role in Pollen Germination

Plants (2022)

Maria Breygina Olga Schekaleva Е. С. Клименко Oksana Luneva

The concept of ROS as an important factor controlling pollen germination and tube growth has become generally accepted in the last decade. However, the relationship between various ROS and their significance for the success of in vivo germination and fertilization remained unexplored. For the present study, we collected Nicotiana tabacum stigma exudate on different stages of stigma maturity before and after pollination. Electron paramagnetic resonance (EPR) and colorimetric analysis were used to assess levels of O•2- and H2O2 on stigma. Superoxide dismutase activity in the stigma tissues at each stage was evaluated zymographically. As the pistil matured, the level of both ROS decreased markedly, while the activity of SOD increased, and, starting from the second stage, the enzyme was represented by two isozymes: Fe SOD and Cu/Zn SOD, which was demonstrated by the in-gel inhibitory analysis. Selective suppression of Cu/Zn SOD activity shifted the ROS balance, which was confirmed by EPR. This shift markedly reduced the rate of pollen germination in vivo and the fertilization efficiency, which was estimated by the seed set. This result showed that hydrogen peroxide is a necessary component of stigma exudate, accelerates germination and ensures successful reproduction. A decrease in O•2- production due to NADPH oxidase inhibition, although it slowed down germination, did not lead to a noticeable decrease in the seed set. Thus, the role of the superoxide radical can be characterized as less important.

Exudate

Stigma

10.3390/plants11070993

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Hydrogen peroxide in tobacco stigma exudate affects pollen proteome and membrane potential in pollen tubes

Plant Biology (2021)

Maria Breygina Е. С. Клименко Е. С. Шилов Alexandra Podolyan Anna Mamaeva

ROS are known to be accumulated in stigmas of different species and can possibly perform different functions important for plant reproduction. Here we tested the assumption that one of their functions is to control membrane potential and provoke synthesis of unique proteins in germinating pollen. We used spectrofluorometry and spectrophotometry to detect H2 O2 in stigma exudate, quantitative fluorescent microscopy of pollen tubes and flow cytometry of pollen protoplasts to reveal effects on membrane potential, and a label-free quantification approach to study pollen proteome changes after H2 O2 treatment. We found that in both growing pollen tubes and pollen protoplasts exudate causes plasmalemma hyperpolarization similar to that provoked by H2 O2 . This effect is abolished by catalase treatment and the ROS quencher, MnTMPP. Inhibitory analysis indicates probable participation of Ca2+ - and K+ -conducting channels in the observed hyperpolarization. For a deeper understanding of pollen response, we analysed proteome alterations in H2 O2 -treated pollen grains. We found 50 unique proteins and 20 differently accumulated proteins that are mainly involved in cell metabolism, energetics, protein synthesis and folding. Observed hyperpolarization and proteome alterations agree well with previously reported stimulation of pollen germination by H2 O2 and sensitivity of Ca2+ - and K+ -conducting channels to this ROS. Thus, H2 O2 is one of the active substances in tobacco stigma exudate that stimulates various physiological processes in germinating pollen.

Exudate

Stigma

Proteome

10.1111/plb.13255

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ХАРАКТЕРИСТИКА ИМПОРТА И ЭКСПОРТА ДНК В МИТОХОНДРИЯХ КАРТОФЕЛЯ (SOLANUM TUBEROSUM) С ИСПОЛЬЗОВАНИЕМ МЕТОДА КОЛИЧЕСТВЕННОЙ ПЦР

Биологические мембраны: Журнал мембранной и клеточной биологии (2011)

Е. С. Клименко Ю. М. Константинов Pavel P. Laktionov В А Милейко Evgeny S. Morozkin

С использованием метода количественной ПЦР изучены особенности транспорта ДНК в изолированные митохондрии клубней картофеля (Solanum tuberosum). Показано, что митохондриальный транспорт рекомбинантной плазмиды pBluescript IIKS (+) размером 3050 п.н. и фрагмента размером 89 п.н. повтора LINE1 человека осуществляется, по всей вероятности, при помощи разных мембранных механизмов. В то же время независимо от длины полинуклеотидной цепи транспорт ДНК в митохондрии является обратимым процессом.

Solanum tuberosum

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Dynamics of ROS production, SOD, POD and CAT activity during stigma maturation and pollination in Nicotiana tabacum and Lilium longiflorum

Plant Biology (2024)

Olga Schekaleva Oksana Luneva Е. С. Клименко S. Shaliukhina Maria Breygina

Abstract Reactive oxygen species (ROS) are a key regulator of physiological processes in pollen grains, and an essential component of stigma exudate. The mechanisms of this redox‐based regulatory system and its features in different plant groups are still unclear. For two species from different families (tobacco and lily), the dynamics of total ROS, O • 2 − generation, and H 2 O 2 concentration in stigma exudate were examined using EPR spectroscopy and quantitative colorimetric analysis. Dynamics of all major enzymes of redox homeostasis were analysed using native electrophoresis and zymography for four stages of stigma development, before and after pollination. There were completely different patterns of ROS production and interconversion in the two species. In tobacco, the initially high level of ROS generation decreased before pollination but remained high. There was no CAT activity in fresh stigma tissues, which apparently contribute to the high level of H 2 O 2 . Lilium had peak O • 2 − generation at the fertile stage and high activity of H 2 O 2 ‐reducing enzymes, including CAT, hence, H 2 O 2 level remained relatively low. We suggest that Lilium pollen germination is largely controlled by the SOD radical, while in Nicotiana H 2 O 2 is the main form of ROS in the stigma.

Lilium

Exudate

Stigma

10.1111/plb.13677

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Citations (2)

Adaptation of the technique of determination of mitochondrial membrane potential change in C2C12 cells using the ready set “Mitochondrial Membrane Potential Kit” (Sigma-Aldrich)

Translational Medicine (2024)

Yulia Vlasova Е. С. Клименко К. С. Сухарева L. S. Gavrilova Anna Kostareva

Background. We used the “Mitochondrial Membrane Potential Kit” (Sigma-Aldrich) to detected MMP but encountered difficulties by applied this kit because in manual there not JC-10 concentration and it not allowing for account cell type, size, density, differences in incubation time for different cell cultures. Objective. Adaptation of the method for determining MMP (mitochondrial membrane potential) in C2C12 cells using in microplate reader and electron microscope. Design and methods. MMP in C2C12 cells was measured by two way, using fluorescence microscopy (Zeiss, Zen program) and using a plate fluorimeter (CLARIOstar (BMG LABTECH). JC-10 and TMRE dyes (Sigma-Aldrich) were used as fluorescent probes. Results. Optimal conditions for detection changes in mitochondrial membrane potential in C2C12 cells were selected. 100- fold dilution of the dye JC-10 (Dye Loading Solution) and replacement of the manufactured buffer to PBS led to repeatability and reproducibility results. Conclusion. When using ready-made kits for measuring MMP, the method proposed by the manufacturer may not be suitable for the selected cell line. In our study to mouse myoblasts of the C2C12 line, a dilution of the dye for loading was required 100 times compared to that recommended by the manufacturer.

C2C12

Sigma

10.18705/2311-4495-2024-11-1-45-54

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