Шпаковский Георгий Вячеславович

Личная информация

Работал в Национальных институтах здоровья США (NIH, 1990–1993), Исследовательском центре Комиссариата атомной энергии Франции в Сакле (CEA-Saclay, 1994–1995), приглашённым профессором Университета Луи Пастера в Страсбурге, Франция (ULP, 1997–1999).

Образование

Период обученияСтрана, городУчебное заведениеДополнительная информация
1973–1978 Россия, Москва Московский государственный университет имени М.В. Ломоносова (МГУ), биологический факультет, кафедра молекулярной биологии Диплом биолога по специальностям молекулярная биология и биохимия (с отличием)
1985 Россия, Москва Институт биоорганической химии им. М.М. Шемякина АН СССР (ИБХ) Присуждена учёная степень кандидата биологических наук
2002 Россия, Москва Институт биоорганической химии им. академиков М.М. Шемякина и Ю.А. Овчинникова РАН (ИБХ) Присуждена учёная степень доктора биологических наук

Премии и заслуги

Лауреат премии ЛКСМ Белоруссии в области науки и техники (1990 г.), премии I степени журнала «Биоорганическая химия» за 1997 г. и Главной премии МАИК «Наука» за 1998 г. по группе биологических наук.

Членство в научных обществах

Член редколлегии журнала Biology Direct (www.biology-direct.com).

Избранные публикации

  1. Tsvetkov V.B., Zatsepin T.S., Belyaev E.S., Kostyukevich Y.I., Shpakovski G.V., Podgorsky V.V., Pozmogova G.E., Varizhuk A.M., Aralov A.V. (2018). i-Clamp phenoxazine for the fine tuning of DNA i-motif stability. Nucleic Acids Res. 46 (6), 2751–2764 [+]

    Non-canonical DNA structures are widely used for regulation of gene expression, in DNA nanotechnology and for the development of new DNA-based sensors. I-motifs (iMs) are two intercalated parallel duplexes that are held together by hemiprotonated C-C base pairs. Previously, iMs were used as an accurate sensor for intracellular pH measurements. However, iM stability is moderate, which in turn limits its in vivo applications. Here, we report the rational design of a new substituted phenoxazine 2'-deoxynucleotide (i-clamp) for iM stabilization. This residue contains a C8-aminopropyl tether that interacts with the phosphate group within the neighboring chain without compromising base pairing. We studied the influence of i-clamp on pH-dependent stability for intra- and intermolecular iM structures and found the optimal positions for modification. Two i-clamps on opposite strands provide thermal stabilization up to 10-11°C at a pH of 5.8. Thus, we developed a new modification that shows significant iM-stabilizing effect both at strongly and mildly acidic pH and increases iM transition pH values. i-Clamp can be used for tuning iM-based pH probes or assembling extra stable iM structures for various applications.

    ID:2074
  2. Shpakovski G.V., Spivak S.G., Berdichevets I.N., Babak O.G., Kubrak S.V., Kilchevsky A.V., Aralov A.V., Slovokhotov I.Y., Shpakovski D.G., Baranova E.N., Khaliluev M.R., Shematorova E.K. (2017). A key enzyme of animal steroidogenesis can function in plants enhancing their immunity and accelerating the processes of growth and development. BMC Plant Biol. 17 (Suppl 1), 189 [+]

    The initial stage of the biosynthesis of steroid hormones in animals occurs in the mitochondria of steroidogenic tissues, where cytochrome P450SCC (CYP11A1) encoded by the CYP11A1 gene catalyzes the conversion of cholesterol into pregnenolone - the general precursor of all the steroid hormones, starting with progesterone. This stage is missing in plants where mitochondrial cytochromes P450 (the mito CYP clan) have not been found. Generating transgenic plants with a mitochondrial type P450 from animals would offer an interesting option to verify whether plant mitochondria could serve as another site of P450 monooxygenase reaction for the steroid hormones biosynthesis.

    ID:1982
  3. Zlobin I.E., Kartashov A.V., Shpakovski G.V. (2017). Different roles of glutathione in copper and zinc chelation in Brassica napus roots. Plant Physiol. Biochem. 118, 333–341 [+]

    We investigated the specific features of copper and zinc excess action on the roots of canola (Brassica napus L.) plants. Copper rapidly accumulated in canola root cells and reached saturation during several hours of treatment, whereas the root zinc content increased relatively slowly. Excessive copper and zinc entry inside the cell resulted in significant cell damage, as evidenced by alterations in plasmalemma permeability and decreases in cellular enzymatic activity. Zinc excess specifically damaged root hair cells, which correlated with a pronounced elevation of their labile zinc level. In vitro, we showed that reduced glutathione (GSH) readily reacted with copper ions to form complexes with blocked sulfhydryl groups. In contrast, zinc ions were ineffective as glutathione blockers, and glutathione molecules did not lose their specific chemical activity in the presence of Zn2+ ions. The effect of copper and zinc excess on the glutathione pool in canola root cells was analysed by a combination of biochemical determination of total and oxidized glutathione contents and fluorescent staining of free reduced glutathione with monochlorobimane dye. Excess copper led to dose-dependent diminution of free reduced glutathione contents in the root cells, which could not be explained by the loss of total cellular glutathione or its oxidation. In contrast, we observed little effect of much higher intracellular zinc concentrations on the free reduced glutathione content. We concluded that GSH plays an important role in copper excess, but not zinc excess chelation, in canola root cells.

    ID:1981
  4. Varizhuk A.M., Zatsepin T.S., Golovin A.V., Belyaev E.S., Kostyukevich Y.I., Dedkov V.G., Shipulin G.A., Shpakovski G.V., Aralov A.V. (2017). Synthesis of oligonucleotides containing novel G-clamp analogue with C8-tethered group in phenoxazine ring: Implication to qPCR detection of the low-copy Kemerovo virus dsRNA. Bioorg. Med. Chem. 25 (14), 3597–3605 [+]

    Nowadays modified oligonucleotides are widely used in diagnostics and as novel therapeutics. Introduction of modified or unnatural residues into oligonucleotides allows fine tuning of their binding properties to complementary nucleic acids and leads to improved stability both in vitro and in vivo. Previously it was demonstrated that insertion of phenoxazine nucleotides with various groups in C9-position into oligonucleotides leads to a significant increase of duplex stability with complementary DNA and RNA. Here the synthesis of a novel G-clamp nucleoside analogue (G(8AE)-clamp) bearing 2-aminoethyl tether at C8-atom is presented. Introduction of such modified residues into oligonucleotides lead to enhanced specificity of duplex formation towards complementary DNA and RNA targets with increased thermal and 3'-exonuclease stability. According to CD-spectroscopy studies G(8AE)-clamp does not substantially disrupt helix geometry. Primers containing G(8AE)-clamp demonstrated superior sensitivity in qPCR detection of dsRNA of Kemerovo virus in comparison to native oligonucleotides.

    ID:1819
  5. Shematorova K., Slovokhotov Y.u., Khaliluev R., Berdichevets N., Baranova N., Babak G., Shpakovski D.G., Spivak S.G., Shpakovski G.V. (2014). Mitochondria as a Possible Place for Initial Stages of Steroid Biosynthesis in Plants. Journal of Stress Physiology & Biochemistry 10 (4), 85–97 [+]

    With the aim of thorough comparison of steroidogenic systems of plants and animals, transgenic plants of Solanaceae family expressing CYP11A1 cDNA encoding cytochrome P450SCC of mammalian mitochondria were further analysed. Positive effect of CYP11A1 on resistance of the transgenic tobacco plants to the infection by fungal phytopathogene Botrytis cinerea was for the first time detected. Subtle changes in mitochondria of the transgenic Nicotiana tabacum plants expressing mammalian CYP11A1 cDNA were demonstrated by transmissive electron microscopy. The main components of the electron transfer chain of plant mitochondria were for the first time cloned and characterized. It was established that plants from the Solanacea family (tomato, tobacco and potato) contain two different genes with similar exon-intron structures (all contain 8 exons) encoding mitochondrial type ferredoxins (MFDX), and one gene for mitochondrial ferredoxin reductase (MFDXR). The results obtained point out on profound relatedness of electron transfer chains of P450-dependent monooxygenases in mammalian and plant mitochondria and support our previous findings about functional compatability of steroidogenic systems of Plantae and Animalia.

    ID:1493
  6. Khaliluev M.R., Shpakovski G.V. (2013). Genetic engineering strategies for enhancing tomato resistance to fungal and bacterial pathogens. Russian Journal of Plant Physiology 60 (6), 721–732 [+]

    The classification and detailed overview of the currently known effective strategies used to increase the resistance of tomato (Solanum lycopersicum L., syn. Lycopersicon esculentumMill.) plants to infectious fungal and bacterial diseases by genetic engineering approaches are presented. Modern data on the mechanisms of the protective effect of heterologous genes on the enhancement of transgenic tomato resistance to fungal and bacterial pathogens are discussed.

     
     
    ID:2079
  7. Shematorova E.K., Shpakovski D.G., Shpakovski G.V. (2013). [Novel complexes of gene expression and their role in the appearance and evolution of the genus Homo]. Tsitologiia 55 (3), 172–7 [+]

    Using genetic (yeast two-hybrid system) and biochemical (co-precipitation of proteins from cellular lysates) approaches, we have performed a whole-genome wide search for interacting partners of the previously described by us variants of hRPB11 subunit of human RNA polymerase II - hRPB1 1balpha, hRPB11calpha and hRPB1 1bbeta, hRPB 11cbeta - in fetal brain and Jurkat cell line libraries. In consequence, the main spectrum of the protein partners of these human specific isoforms of the RNA polymerase II subunit hRPB 11 (POLR2J) has been established. Functional characteristics of the uncovered protein partners of hRPB 11balpha and hRPB 11calpha isoforms clearly indicate that these isoforms, similarly to the main (major) subunit hRPB11a, are components of the distinct transcription complexes participating not only in the transcription of the specific DNA matrices, but involving also in the later stages of mRNA biogenesis. The RNA polymerase I-III common subunit hRPB6 (POLR2F) and basal component of the exon-exon junction complex Y14 (RBM8A) have been found among the protein partners of the isoforms hRPB 11bbeta and hRPB 11cbeta together with a number of proteins involved in the biogenesis of microRNAs, including a novel, not previously described variant of the microRNA processing nuclease DROSHA, which indicates the existence of a special coordination between processes of transcription and RNA interference in the nuclei of human cells.

    ID:1492
  8. Proshkin S.A., Shematorova E.K., Souslova E.A., Proshkina G.M., Shpakovski G.V. (2011). A minor isoform of the human RNA polymerase II subunit hRPB11 (POLR2J) interacts with several components of the translation initiation factor eIF3. Biochemistry Mosc. 76 (8), 976–80 [+]

    Using the yeast two-hybrid (YTH) system we have uncovered interaction of the hRPB11cα minor isoform of Homo sapiens RNA polymerase II hRPB11 (POLR2J) subunit with three different subunits of the human translation initiation factor eIF3 (hEIF3): eIF3a, eIF3i, and eIF3m. One variant of eIF3m identified in the study is the product of translation of alternatively spliced mRNA. We have named a novel isoform of this subunit eIF3mβ. By means of the YTH system we also have shown that the new eIF3mβ isoform interacts with the eIF3a subunit. Whereas previously described subunit eIF3mα (GA17) has clear cytoplasmic localization, the novel eIF3mβ isoform is detected predominantly in the cell nucleus. The discovered interactions of the hRPB11cα isoform with several hEIF3 subunits demonstrate a new type coordination between transcription and the following (downstream) stages of gene expression (such as mRNA transport from nucleus to the active ribosomes in cytoplasm) in Homo sapiens and point out the possibility of existence of nuclear hEIF3 subcomplexes.

    ID:1486
  9. Shematorova E.K., Shpakovski D.G., Shpakovski G.V. (2010). [PSM2 and POLR2J gene families as molecular markers of the higher primate evolution]. Genetika 46 (9), 1254–7 [+]

    We have studied the molecular evolution of two gene families specific for primates: POLR2J of the transcription system and PMS2 of the MMR repair system. The appearance and improvement of the genetic structure in each of the families was shown to strongly correlate with the main stages of the higher primates biological evolution. Our results indicate that the PSM2 and POLR2J genes can serve as helpful and reliable molecular markers of anthropogenesis.

    ID:1491
  10. Spivak S.G., Berdichevets I.N., Iarmolinskiĭ D.G., Maneshina T.V., Shpakovski G.V., Kartel N.A. (2009). [Construction and characteristics of transgenic tobacco Nicotiana tabacum L. plants expressing CYP11A1 cDNA encoding cytochrome P450scc]. Genetika 45 (9), 1217–24 [+]

    In steroidogenic animal tissues cytochrome P450scc catalizes the conversion of cholesterol into pregnenolone, a common metabolic precursor of all steroid hormones. To study the possibility of functioning of mammalian cytochrome P450scc in plants and the mechanism of its integration in the plant steroidogenic system, transgenic plants of tobacco Nicotiana tabacum L. were developed carrying cDNA of CYP11A1 encoding cytochrome P450scc of bovine adrenal cortex. Pregnenolone, a product of the reaction catalyzed by cytochrome P450scc, was discovered in the steroid-containing fraction of transgenic plants. Transgenic plants are characterized by a reduced period of vegetative development (early flowering and maturation of bolls) and increased productivity. The contents of soluble protein and carbohydrates in leaves and seeds of transgenic plants are essentially higher than the contents of these components in leaves and seeds of control plants.

    ID:1488
  11. Spivak S.G., Berdichevets I.N., Litvinovskaia R.P., Drach S.V., Kartel N.A., Shpakovski G.V. (2009). [Characteristics of steroid metabolism in transgenic Nicotiana tabacum plants bearing the CYP11A1 cDNA of cytochrome P450(SCC) from the bovine adrenal cortex]. Bioorg. Khim. 36 (2), 241–50 [+]

    In the mitochondria of animal steroidogenic tissues, cytochrome P450(SCC), encoded by the CYP11A1 gene, catalyzes the conversion of cholesterol into pregnenolone - the general precursor of all steroid hormones. In this work, we study the steroid metabolism in transgenic tobacco plants carrying the CYP11A1 cDNA cytochrome P450(SCC)from the bovine adrenal cortex. The transgenic plants under investigation markedly surpass the control wild-type plants by size and are characterized by a shortened period of vegetative growth (by rapid flowering); their leaves contain pregnenolone - the product of a reaction catalyzed by cytochrome P450(SCC). The level of progesterone in transgenic tobacco leaves is higher than in the control plants of the wild type. The seeds of the transgenic plants contain less (24R)-brassinosteroids than the wild-type tobacco plants. The results obtained indicate that the synthesis of an active P450(SCC) cytochrome in transgenic Nicotiana tabacum plants has a profound effect on steroid metabolism and is responsible for the specific phenotypic features of transgenic plants bearing CYP11A1 cDNA.

    ID:1490
  12. Shpakovski D.G., Shematorova E.K., Shpakovski G.V. (2006). Human PMS2 gene family: origin, molecular evolution, and biological implications. Dokl. Biochem. Biophys. 408 (5), 175–179 [+]

    Проведен филогенетический анализ генов семейства PMS2 приматов и уточнена их классификация. Установлено, что в эволюции этого генного семейства имеются стадии, специфичные для человека. Предложена гипотеза о трёхкомпонентной белковой системе, продуцируемой PMS2-подобными генами Homo sapiens.

    ID:61
  13. Proshkina G.M., Shematorova E.K., Proshkin S.A., Zaros C., Thuriaux P., Shpakovski G.V. (2006). Ancient origin, functional conservation and fast evolution of DNA-dependent RNA polymerase III. Nucleic Acids Res. 34 (13), 3615–24 [+]

    Определены основные этапы эволюции ядерных РНК-полимераз I—III эукариот. Показано, что состоящий из трёх РНК-полимераз базовый аппарат транскрипции ядерных организмов возник очень давно и характерен для представителей всех основных царств (таксономических супергрупп) эукариот. В ходе дальнейшей эволюции трёх ферментов транскрипции состоящий из двенадцати субъединиц кор РНК-полимераз оставался консервативным, а субъединицы, специфичные для каждого из трёх типов полимераз, быстро дивергировали. В составе 12-субъединичного кóра РНК-полимеразы III впервые обнаружены три высоко консервативных домена, специфичных для этого фермента транскрипции.

    ID:60
  14. Shpakovski D.G., Shematorova E.K., Shpakovski G.V. (2004). New genes on human chromosome 7: bioinformatic analysis of a gene cluster from the POLR2J family. Bioorg. Khim. 30 (6), 621–5 [+]

    В составе хромосомы 7 человека выявлены четыре независимых гена, кодирующих различные варианты субъединицы hRPB11 РНК-полимеразы II Homo sapiens. Установлено, что при экспрессии четырёх генов POLR2J человека могут синтезироваться, по крайней мере, 14 видов зрелых, кодирующих слегка различные изоформы hRPB11, мРНК, 11 из которых охарактеризованы. Предложена и обоснована схема происхождения множественных генов семейства POLR2J путём трёх увеличивающихся в размерах дупликаций, позволяющая сделать ряд интересных наблюдений о путях эволюции отдельных человеческих генов и о механизмах генерирования белкового разнообразия у высших эукариот.

    ID:59
  15. Wood V., Gwilliam R., Rajandream M.A., Lyne M., Lyne R., (> 100 authors here) , Paulsen I., Potashkin J., Shpakovski G.V., Ussery D., Barrell B.G., Nurse P., Cerrutti L. (2002). The genome sequence of Schizosaccharomyces pombe. Nature 415 (6874), 871–80 [+]

    При активном участии лаборатории завершено секвенирование и аннотация (см. также: Биоорган. химия, 1999, 25: 450–463) генома делящихся дрожжей Schizosaccharomyces pombe. В рамках международного проекта по полному секвенированию генома Schizosaccharomyces pombe в трёх локусах восстановлена непрерывность нуклеотидной последовательности хромосомы I: три изолированных в нашей лаборатории клона (pYUK71, pYUL23 и pYUG7) вошли в список клонов, составляющих полную физическую карту генома делящихся дрожжей. Впервые российская лаборатория была отмечена как полноправный участник проекта полного секвенирования генома эукариотического организма.

    ID:58
  16. Shpakovski G.V., Gadal O., Labarre-Mariotte S., Lebedenko E.N., Miklos I., Sakurai H., Proshkin S.A., Van Mullem V., Ishihama A., Thuriaux P. (2000). Functional conservation of RNA polymerase II in fission and budding yeasts. J. Mol. Biol. 295 (5), 1119–27 [+]

    Ранее нами была впервые продемонстрирована функциональная взаимозаменяемость in vivo отдельных субъединиц эукариотических РНК-полимераз между эволюционно далекими организмами (Г. В. Шпаковский и др.: Gene, 1994, 147: 63–69; Mol. Cell. Biol., 1995, 15: 4702–4710) и и установлено функциональное родство малых субъединиц РНК-полимераз архей и эукариот (Биоорган. химия, 1997, 23: 110–117; J. Biol. Chem., 1999, 274: 8421–8427). В данной итоговой работе было проведено первое систематическое исследование взаимозаменяемости in vivo всех двенадцати субъединиц ядерной РНК-полимеразы II между эволюционно далекими видами дрожжей, Schizosaccharomyces pombe и Saccharomyces cerevisiae, а также завершены клонирование и функциональная характеристика всех компонентов РНК-полимеразы II делящихся дрожжей.

    ID:57