Registration Number
0225U004155,
(0120U102238)
, R & D reports
Title
Structural and functional studies of translation elongation factors of higher eukaryotes
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Дослідження функціональної ролі метилування специфічних лізинових залишків ізоформ eEF1A1 і eEF1A2 фактора елонгації трансляції 1А
Head
Negrutskyii Borys S., Доктор біологічних наук
Registration Date
20-11-2025
Organization
Institute of Molecular Biology and Genetics of NAS of Ukraine
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The aim of this study is to analyze the spatial organization and function of mammalian translation elongation factor 1 (eEF1A and the multi-protein eEF1B complex). To achieve this, the following tasks are planned. First, to obtain experimental data that will allow us to build atomic models of the spatial organization of the subunits of the eEF1B translation complex and the architecture of the complex as a whole. Second, to conduct experimental studies and to build the model of spatial organization of isoform 1 of elongation factor 1A based on them. Third, to determine the potential functional significance of methylation of lysine residues in the molecule of elongation factor 1A (eEF1A), in particular, to test the hypothesis experimentally that the methylation of some lysine residues in eEF1A may affect the interaction of this protein with partners , such as the subunits of the eEF1B complex, which may reveal the molecular mechanism of the recently discovered effect of eEF1A methylation on oncogenic cell transformation. Fourthly, describe the molecular mechanisms of action of novel eEF1A inhibitors with potential antitumor effect.
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Objects of study – the human translation factor complex eEF1B, the translation elongation factor eEF1A, its isoforms eEF1A1 and eEF1A2, and the elongation factors eEF1Bα, eEF1Bβ, and eEF1Bγ.
Objective of the work – to elucidate the spatial organization of translation complexes and their components, and to investigate the potential role of post-translational modifications in the functioning of translation factors.
Research methods – hydrogen–deuterium exchange followed by mass spectrometry, analytical ultracentrifugation, protein mutagenesis, chromatographic purification of individual proteins, analytical gel filtration, bioluminescence resonance energy transfer (BRET) system, Western blotting, co-immunoprecipitation, protein co-precipitation, confocal microscopy, mass spectrometry, polymerase chain reaction (PCR), molecular biology methods for DNA manipulation, kinetic methods, bioinformatic analysis, molecular modeling, and molecular docking.
It was found that partial or complete substitution of methylated lysine residues with arginine in the oncogenic isoform of the translation elongation factor eEF1A2 does not affect its interaction with the nucleotide exchange factor eEF1B, which allows this interaction to be excluded from the potential mechanisms of carcinogenesis induced by eEF1A2 methylation. At the same time, a markedly reduced interaction with eEF1B was observed for the eEF1A1 isoform in which all five methylated lysine residues were replaced by arginine. Thus, one of the functional roles of eEF1A1 methylation may be to modulate the efficiency of nucleotide exchange in this molecule.
Product Description
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Novosylna Oleksandra V.
Hanna V. Yelska
Porublova Larysa V.
Luk`yanenko Ganna I.
Tetiana V. Bondarchuk
Вячеслав Ф. Шалак
Oleksandr Y. Tsuvariev
Kolodka Nazar Tarasovych
Sofia Gryciv
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2025-11-20
