Historical information about the laboratory
Laboratory of protein and nucleic acids research (LPNAR) founded in 1968.
Academician Murat Abenovich Aitkhozhin is founder of LPNAR as well as Institute of molecular biology and biochemistry (1983).
Professor Iskakov B. K. is Head of laboratory since 1988.
Iskakov Bulat Kudaibergenovich
Doctor of Biological Sciences, Professor
As member of a group of young scientists, Iskakov B. K. was awarded the title of Laureate of the Lenin Komsomol prize in the field of science and technology in 1979. Iskakov B. K. defended his doctoral dissertation in the dissertation Council at Lomonosov Moscow State University on “Investigation of molecular mechanisms and regulation of protein biosynthesis in plants”, specialty 03.00.03 “molecular biology” in 1997. Iskakov B.K.was awarded the title of Professor in the specialty “Biology” in 2002.
Under the leadership of Iskakov B. K., it was established and improved the biotechnology to obtain drought-resistant transgenic plants (potatoes, tobacco), as well as plants resistant to phytopathogenic viruses, using approaches antisense RNA and RNA interference, for the first time in Kazakhstan. It was established also that specific mechanisms of translational control, different from animals, can function in plant cells, which can provide the initiation of translation without the participation of some protein factors.
Prof. Iskakov B. K. led research projects in the framework of many international projects financed by grants from foreign funds INTAS, US AID, INCO COPERNICUS, Royal Society, SCOPUS from 1994 to 2004. These projects have been successfully implemented together with scientists from Switzerland, Germany, Great Britain, Russia and Israel. Iskakov B. K. is the author of more than 200 publications. His fundamental research of molecular mechanisms of transmission regulation in plants in normal and under the influence of adverse environmental factors is a priority and meets the requirements of the international standard. This is evidenced by publications in scientific international journals: FEBS Letters, Biochimica et Biophysica Acta, European Journal of Biochemistry, Biochimie, Plant Science, Nuclear Acids Research, Molecular biology, Biochemistry, Plant Physiology, Biopolymers and Cell. Prof. Iskakov B. K. acted as a Guest speaker at many international conferences, and has a h-index 5 and more than 120 citations.
He developed a model curriculum “Genetic engineering” and teaches students of the faculty of biology and biotechnology of al-Farabi KazNU this course of lectures as well as other special courses at the Department of molecular biology and genetics from 1998 to the present time. 6 candidate, 1 doctoral and 1 PhD dissertation were completed and defended under the leadership of Prof. Iskakov B. K. He is the scientific supervisor of PhD doctoral students.
The lab team consists of 15 people: Doctor of Biological Sciences – 1; candidates of biological Sciences – 5; PhD – 1; masters – 5; PhD doctoral student – 1; young scientists up to 35 years – 6. The research team of LPNAR has a high theoretical and methodological level. .Many employees have passed training courses in the leading centers of molecular biology of plants, including training in Switzerland, the UK, Israel, Germany, France, Hungary
Main areas of research
- investigation of molecular mechanisms and regulation of protein biosynthesis in plants;
- the study of the mechanisms regulating the activity of protein factors of translation as well as of discrete fragmentation of the 18S rRNA;
- study the structural elements of the RNA that determine the effectiveness of their translation;
- study of plant virus genome’ structure and expression, RNA interference;
- production of transgenic plants resistant to phytopathogenic viruses (PVY, PVM, PVS), adverse environmental factors (drought,salinization, extreme temperatures);
obtaining transgenic plants producing recombinant vaccines, proteins for medical purposes.
Current research projects (2018-2020):
AP05130800 “Identification and study of translation amplifiers, universal for plant and bacterial gene expression systems”, supervisor –Zhigailov A.V.
АР05131133 “Detection of S protein of potato virus that suppress the process of RNA interference of host cells, with the aim of investigating the molecular mechanisms of interaction between virus and plants and the recovery of the viral material”, supervisor –Karpova O.V.
AP05132066 “Development of technology for expression of recombinant antigens of the virus of sheep pox in transplastomic plants”, supervisor –Stanbekova G. E.
Methods of research:
Methods of molecular biology and genetic engineering to create structures that allow to express recombinant genes.
Amplification by RT-PCR, in vitro-mutagenesis, isolation and analysis of PAAG electrophoresis and radioautography, immunobloting using specific antibodies.
Scientific achievements of the laboratory
- It was shown that the affinity factor of initiation of translation of plants peIF2 to GDP (KdGDP = 150 nM) just in 10 times greater than his affinity for GTP (KdGTP = 1500 nM), whereas for the same factor from mammalian cells (meIF2), this excess is 100 times or more. For the first time stated that the exchange of GDP for GTP at the factor peIF2 plants can occur without the participation of an additional factor eIF2В, which is necessarily required to exchange guanyl nucleotides of the animal factor meIF2. It was first postulated that in plant cells the exchange of GDP to GTP in peif2 factor can occur regardless of whether peIF2 factor is phosphorylated or not. (Shaikhin S. M., Smailov S. K., Lee A. V., Kozhanov E. V., Iskakov B. K. Biochimie, 1992, vol. 74, pp. 447-454). Our results and conclusions were fully confirmed in the works of other groups of researchers and have since been cited in all scientific reviews and articles on protein synthesis in plants. (IF 3.188).
- It was established that in plant cells no phosphoprotein-kinase that specific phosphorylates the protein elongation factor translation 2 (peEF2). These data suggest that unlike animals plants lack the mechanism of regulation of activity of key elongation factor of translation peEF2 by reversible phosphorylation. ( Smailov S. K., Lee A.V., Iskakov B. K. FEBS Letters, 1993, vol. 321, pp. 219-223). Subsequently, our results have been recognized worldwide and cited in scientific reviews and articles on the biosynthesis of protein in plants. (IF 2.999).
- We studied the mechanism of phenolic compounds from Alhagi kirgisorum S., i.e polypropanthocianidine (PPA) as the inhibitor of the phenolic compounds. It is shown that PPA in concentrations of 1-10 mm is specifically associated with the initiation factor eIF2 and inhibits its activity, which is accompanied by blocking of protein synthesis. Thus, PPA is a convenient tool for studying the mechanisms of translation initiation in eukaryotic cells. This work was carried out in conjunction with the laboratory of Prof. Kunaeva R. M., as well as scientists from Russia and the United States. The results were published in 2 international journals ( Smailov S. K., Mukhamedzhanov B. G., Lee A. V., Iskakov B. K., Denisenko, O. N. FEBS letters, 1991, vol. 275, pp. 99-101, IF 2.999; Kudlicki W., Picking W., Kramer G., Hardesty B., Smailov S., Mukhamedzhanov B., Lee A., Iskakov B. European journal of biochemistry, 1991, vol. 197, pp. 623-629. IF 4,530).
- We found and then studied 45S ribonucleoprotein (RNP) complexes in the cells of wheat germ. It was found that 45S RNPs represent preinitialize translational complexes that contain the small (40S) ribosomal subunit, protein factors of translation initiation and mRNA in RNP-form. As part of 45S RNP complexes, we first discovered a new small RNA (5,3S RNA) with a length of 134 nucleotides. (Iskakov B.K., Madin K.I. Plant Science, 1994, vol. 96, pp. 99-108. IF 3.712).
- A new cap-independent mechanism of mRNA binding to 40S ribosomal subunit during translation initiation was established for plant objects for the first time. This mechanism consists in the complementary interaction of a 5’-untranslated region of mRNA (5’-UTR) with the central domain of 18S rRNA. It is experimentally proved that the increase in the level of complementarity in 5’-UTR to this 18S rRNA site leads to a multiple increase in the efficiency of mRNA translation. The results are not only fundamental, but also of applied importance, because they allow us to artificially construct mRNAs with very high translational activity. Such highly active mRNAs are necessary in cell-free protein synthesis technology as well as in genetic engineering to produce transgenic plants producing valuable proteins. (Akbergenov R.Z., Zhanybekova S.S., Polimbetova N.S., Madin K.I., Hohn T., Iskakov B.K. Complementary interaction between the central domain of 18S rRNA and the 5’ untranslated region of mRNA enhances translation efficiency in plants. In: “Cell-Free Protein Expression”, James R. Swartz (Ed.), ISBN 3-540-05041-8, Springer Verlag Berlin Heidelberg New York, 2003, pp. 199-208. Akbergenov R.Z., Zhanybekova S.S., Kryldakov R.V., Zhigailov A., Polimbetova N.S., Hohn T., Iskakov B.K. ARC-1, a sequence element complementary to an internal 18S rRNA segment, enhances translation efficiency in plants when present in the leader or intercistronic region of mRNAs. Nucleic Acids Research, 2004, vol. 32, No. 1, pp. 239-247. IF 11.237).
- It was found a new small 5,3S in the 40S of ribosomal subunits of wheat germ for the first time. The amount of 5,3S RNA increases in 5 times under temperature stress. It was found that 5,3S RNA is a specific 5’-end fragment of 18S rRNA, which is formed as a result of the action of endogenous nucleases. (Zhanybekova S., Polimbetova N., Nakisbekov N., Iskakov B. Biochemistry (Moscow), 1996, vol. 61, pp. 621-627, IF 1.724; Polimbetova N.S., Zhanybekova S.Sh., Lee A.V., Iskakov B. K. Plant Physiology, 1996, vol. 43, pp. 887-893, IF 0.816).
- Biotechnology for producing transgenic plants resistant to phytopathogenic viruses, drought and cold has been established. Transgenic tobacco and potato plants expressing antisense RNAs complementary to different sites of potato virus Y genomic RNA were obtained. Several lines of transgenic potatoes were submitted for the testing into the Institute of potato and vegetable farming of the Ministry of agriculture. Many transgenic potato lines have shown considerable resistance to potato virus Y and are considered very promising for further breeding process. GM potato lines with multiple resistance to PVY, PVM, PVS viruses were obtained by induction of RNA interference.
- Technologies have been established for the production of transgenic bacteria and plants, as well as transplastomic plants, which produce recombinant vaccine proteins against the sheep pox virus (SPPV), as well as medicine-significant proteins (hAFP).