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Contract number
14.Z50.31.0005
Time span of the project
2014-2016
Head of the laboratory

As of 01.06.2020

18
Number of staff members
47
scientific publications
General information

Name of the project: Mechanisms of kinetochore-dependent formation of microtubes in Drosophila

Strategy for Scientific and Technological Development Priority Level: в


Goals and objectives

Research directions:

- Developing new methods to analyze growth of microtubes directed by kinetochores

- Search for new proteins necessary for growth of microtubes directed by kinetochores and characterization of those proteins

- Functional analysis of highly evolutionary conservative homologs of drosophila proteins controlling mitosis

Project objective: Finding molecular mechanisms of growth of microtubes providing chromosome division in mitosis (division of somatic cells) in a classic genome model object – Drosophila flies


The practical value of the study

  • We have conducted ultra-structural characterization of all the stages of mitosis in S2 drosophila using transmission microscopy. We have found that during mitotic division these cells form unique four-layer nuclear membranes that have never been observed in cells of ther organisms.
  • Our researchers have developed an experimental method of in vivo analysis of tubuline monomer flow velocity in beams of microtubes of spindle apparatus that allows for more precise measurement.
  • The Laboratory has created a theoretical model of interpretation of fluorescence recovery curve after discoloration of mitotic spindle apparatus.
  • Using the RNA interference method we have conducted screening to detect protein factors participating in kinetochore-dependent formation of microtube beams of spindle apparatus. We have isolated a range of proteins that are active in this process (Mast/Orbit, Eb1, Dgt6, Mars/DHURP, Mei38/DTPX2, Asp, Patronin) and detected functional connections between some of them.
  • We have compiled a list of evolutionary conservative genes necessary for formation of spindle apparatus: CLASP1 [Mast/Orbit], ASPM [asp], DLGAP5/HURP [Mars], TPX2 [Mei-38/Ssp1], HAUS6 [Dgt6], KIF18A [Klp67A], PLEKHA7 [Patronin/Ssp4], EIF3E [Int6], TRIM13/RFP2 [Non3], PHF20 [MBD-R2], KANSL3 [Rcd1] и MCRS1 [Rcd5]. These genes are prospective for further research of their expression in tumors, developing methods of diagnostics and therapy of oncological conditions.

Implemented results of research:

We have broadened fundamental knowledge about microtubes comprising the basis of spindle apparatus in all eukaryotic organisms as well as about specific proteins associated with them. This is necessary for understanding molecular mechanisms om the basis of conditions connected with disturbance of cell division, in particular, tumor cells in humans.

Education and career development:

  • 1 candidate dissertation, 1 masters dissertation, 2 bachelor dissertation have been defended.
  • Lectures at scientific schools on molecular and cellular biology organized by the Institute of Molecular and Cellular Biology of the Siberian Department of the Russian Academy of Sciences.
  • Employees of the Laboratory participated in conducting seminars and a case study in biochemistry for students of the Novosibirsk State University.

Organizational and structural changes: We have enhanced equipment of the Institute which broadened capabilities in developing and implementing new prospective research methods.

Other results:

We have organized international conferences:

«Chromosome» (2015, 2018, Russia), «Chromosome and mitosis» (2015–2018, Russia).

Collaborations:

  • Sapienza University of Rome (Italy): joint research of the role of the NSL/KANSL protein complex in mitosis in drosophila and humans
  • Netherlands Cancer Institute (the Netherlands): joint research of impact of local environment of chromatin on functional activity of promoter element in cultivated embryonic stem cells of mice, features of heterochromatin in specialized cellular types of drosophila
  • Albert Einstein Medical Center (USA): joint research of regulatory function and load dynamics in chromatin linker histone H1 during the process of endoreplication in drosophila
  • Ludwig Maximilian University of Munich (Germany): joint research of dynamics of interaction of protein receptor Polycomb with drosophila genome during development of wing imaginal discs

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Renda F., Pellacani C., Strunov A., Bucciarelli E., Naim V., Bosso G., Kiseleva E., Bonaccorsi S., Sharp D.J., Khodjakov A., Gatti M., Somma M.P.
The Drosophila orthologue of the INT6 oncoprotein regulates mitotic microtubule growth and kinetochore structure. PLoS Genetics 13(5): e1006784(2017).
Omelyanchuk L.V., Munzarova A.F.
Theoretical model of mitotic spindle microtubule growth for FRAP curve interpretation. BMC Systems Biology 11(Suppl 1): 3(2017).
Strunov A., Boldyreva L.V., Pavlova G.A., Pindyurin A.V., Gatti M., Kiseleva E.
A simple and effective method for ultrastructural analysis of mitosis in Drosophila S2 cells. MethodsX 3: 551–559(2016).
Munzarova A., Popova J., Razuvaeva A., Shloma V., Gatti M., Omelyanchuk L.
Accurate measurement of poleward microtubule flux in the spindle of Drosophila S2 cells. Cell. Biol. Int., 40: 984–990(2016).
Pavlova G.A., GalimovaYu.A., Popova Yu.V., Munzarova A.F., Razuvaeva A.V., Alekseeva A.L., Berkaeva M.B., Pindyurin A.V., Somma M.P., Gatti M., Renda F.
Factors governing the pattern of spindle microtubule regrowth after tubulin depolymerization //Цитология. 2016. Т. 58. № 4. С. 299–303.
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