Registration Number
0225U004554,
(0121U109973)
, R & D reports
Title
Peculiarities of functional activity of ribosomal protein S6 (S6K1) kinase isoforms and molecular mechanisms of its regulation in normal and pathology
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Аналіз змін в стані ключових сигнальних каскадів залучених до злоякісної трансформації з застосуванням модельних клітин карциноми простати, з диференційною експресією ізоформ S6K1. Дослідження рівня проліферації, міграції, виживаності, впливу специфічних інгібіторів mTOR/S6K, ефективності формування колоній у напіврідкому агарі стабільних клітинних ліній, отриманих на основі клітин карциноми простати з редагованою експресією ізоформ S6K1. Імуногістохімічний аналіз локалізації ER в клітинах з диференційною експресією ізоформ S6K1. Аналіз РНК/білкових комплексів - мРНК кіназ та RBP. Застосування імунопреципітації РНК (RIP) до підтверджених раніше та аналіз розподілу нормальних та мутантних мРНК кіназ (кількісний ПЛР) у визначених клітинах. Аналіз синергізму, антагонізму та компенсаторної дії знайдених RBP специфічних до різних ділянок мРНК S6K.
Head
Filonenko Valerii V., д.б.н.
Registration Date
11-12-2025
Organization
Institute of Molecular Biology and Genetics of NAS of Ukraine
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Determination and characterization of features of functional activity and molecular mechanisms of its regulation of known isoforms of ribosomal protein kinase S6K1 - p55, p60, p70, p85 in normal and malignant transformation of cells.
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The purpose of the work is to determine the features of the functional activity of ribosomal protein kinase S6 -S6K1 isoforms in normal and pathological conditions.
Research methods are the CRISPR/Cas9 genome editing system and shRNA interference, gene expression analysis by real-time PCR, Western blot analysis of protein expression and their phosphorylation, immunocytochemistry and confocal microscopy. The features of the functional activity of ribosomal protein kinase S6 (S6K1) isoforms - p60, p70 and p85 and the molecular mechanisms of their regulation in normal and pathological conditions were determined. Using the CRISPR/Cas9 genome editing system, several stable human breast carcinoma cell lines of different molecular subtypes with differential expression of S6K1 isoforms were created, thanks to which their role in the progression of tumor growth at the level of invasion and metastasis was determined. It was found that the functional activity of S6K1 isoforms in tumors of different molecular subtypes differs significantly, and therefore this should be taken into account in antitumor therapy. A correlation was established in the expression of S6K1 isoforms in tumors of different histotypes according to such indicators as invasiveness and metastasis. A number of post-translational modifications involved in the regulation of the functional activity of the kinase were identified and characterized, including methylation, acetylation and CoAlylation, which determine the subcellular localization and substrate specificity of the kinase. It was established that CoAlylation is a new mechanism of negative regulation of S6K1 activity, which is an alternative to classical dephosphorylation and is inherent not only to S6K1, but also to other kinases such as Aurora A. A connection was revealed between CoAlylation of proteins and pathological human conditions, including neurodegenerative and oncological disease
Product Description
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Yunhin Olha S.
Liliia O. Savinska
Nazarii Y. Hotsuliak
Harifulin Oleh M.
Bdzhola Anna V.
Tykhonkova Iryna
Martsyniuk Marko Ye.
Alla P. Pohribna
Iryna V. Krupska
Vasyl I. Bobyk
Olha H. Vihontina
Oksana М. Malanchuk
Sergiy Palchevskii
Andrii Mazov
Ludmyla Yakovenko
Oleksandr Skorohod
Antonina Khoruzhenko
Ol'ga Nimenko
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2025-12-11
