Laboratory of protein and nucleic acids research

Kakimbek Yegizbayev

5 years ago

A brief history of the Laboratory

Laboratory of protein and nucleic acids research (LPNAR) founded in 1968. A founder and the first Head of the LPNAR is Academician Murat Abenovich Aitkhozhin. From 1988 to 2022 the head of the laboratory was Professor Bulat Kudaibergenovich Iskakov. Since January 2023 Dr. Andrey Zhigailov, has been appointed as a head of the laboratory.

Zhigailov Andrey Viktorovich

Ph.D in biology

Head of the Laboratory of protein and nucleic acids

Phone: +7 (727) 239-05-07

E-mail: andrzhig@gmail.com

Andrey Zhigailov graduated from the biological faculty of al-Farabi Kazakh National University in 2002. He earned his master’s degree in biotechnology in 2004. He completed postgraduate studies at the Institute of molecular biology and biochemistry in 2006. He passed an internship at the Institute of chemical biology and fundamental medicine, Siberian branch of the Russian Academy of Sciences, Novosibirsk, Russia (2006); training on “Fundamentals of sequencing and fragmentary analysis” Warrington, UK (2007); internship on mastering the methods of working with miRNAs at the Institute of botany of Basel, Switzerland (2007). In 2018, he passed advanced training: “Methods and techniques for working with pathogens of especially dangerous infections of II-IV pathogenicity groups” 216 hours at the M. Aikimbaev Kazakh scientific center for quarantine and zoonotic infections in Almaty.

Since 2001 he had been working at the Laboratory of protein and nucleic acids of IMBB as: laboratory assistant – junior researcher – researcher – senior researcher. Between 2016 and 2022 – Leading researcher at the Laboratory of protein and nucleic acids of IMBB. Since January 2023, he has been appointed as a Head of the Laboratory. From 2012 to the present, he has been a scientific expert in reviewing and reviewing scientific projects and reports as part of a comprehensive review conducted by the National center for state scientific and technical expertise JSC.

He took part in 11 scientific projects and grants, in three of them he was a scientific supervisor. Hirsch index: 3, Researcher ID (Web of Science): N-6073-2015, Scopus ID: 6508121286. ORCID: https://orcid.org/0000-0002-9646-033X.

In 2016, he was awarded the “Algys” gratitude of the Ministry of Education and Science of the Republic of Kazakhstan.

The main directions of research activities

- 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.

Research projects:

2022-2024

AP14869357 “Development of the scientific basis of biotechnology for increasing plant productivity through modification of the genes of the ribosomal protein S6”, scientific supervisor – Ph.D. Zhigailov A.V.

2020-2022

AP08855746 “Development of biotechnology for increasing plant productivity by activating the process of ribosome biogenesis” – supervisor – Ph.D., Zhigailov A.V.

2018-2020

AP05130800 “Identification and study of translation amplifiers, universal for plant and bacterial gene expression systems”, scientific 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”, scientific supervisor –Karpova O.V.

AP05132066 “Development of recombinant antigens expression technology of the sheep pox virus in transplastomic plants”, scientific supervisor –Stanbekova G. E.

2015-2017

4538 / GF4 “Development of biotechnology for the creation of genetically modified potato plants with increased resistance to abiotic stresses based on the optimization of the expression of mutated variants of the AteIF2α transgene”, scientific supervisor – Dr. of Biological Sciences. Iskakov B.K.

1920/GF4 “Study of the regulation of mRNA translation in plants through phosphorylation of the alpha subunit of initiation factor 2 (peIF2α)”, scientific supervisor – Ph.D., Zhigailov A.V.

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

1. 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; Times Cited 17).

1. It was found 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; Times Cited 14).

1. For the first time, the mechanism of action of a phenolic compound from camel thorn (Alhagi kirgisorum S.) – polyproanthocyanidin (PPA) as an inhibitor of protein synthesis initiation in plants and animals was studied. It was shown that PPA at concentrations of 1-10 mM specifically binds to 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. The PPA preparation was provided by us for research work to scientists from different countries (USA, Great Britain) and was highly appreciated. This work was carried out in collaboration with the laboratory of professor Rauza Kunaeva, as well as with scientists from Russia and the USA. 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; Times Cited 8) 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; Times Cited 5).

1. For the first time in wheat germ cells, we discovered and then studied 45S ribonucleoprotein (RNP) complexes. 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; Times Cited 4)

1. 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, Times Cited 63)

1. For the first time, a new small 5,3S RNA was found in the 40S ribosomal subparticles of wheat germs, and its properties were studied. The amount of 5,3S RNA increases 5-fold under conditions of temperature stress. It has been established that 5,3S RNA is a specific 5′-terminal fragment of 18S rRNA, which is formed as a result of the action of endogenous nucleases. Published in Russian and domestic journals: Zhanybekova S., Polimbetova N., Nakisbekov N., Iskakov B. (1996) Biochemistry (Moscow), V. 61, P. 621-627; (IF 1.724, Times Cited 3), Polimbetova N.S., Zhanybekova S.Sh., Lee A.V., Iskakov B.K. (1996) Plant Physiology, vol. 43, pp. 887-893; (IF 0.816)

1. 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 for obtaining transgenic bacteria and plants have been established, as well as transplastomic plants, which produce recombinant vaccine proteins against the sheep pox virus (SPPV), as well as medicine-significant proteins (hAFP).