Advances in Chemical PhysicsAdvances in Chemical PhysicsФизиология растений0015-33033034-6126The Russian Academy of Sciences64814710.31857/S0015330323600067QAHCTEЭКСПЕРИМЕНТАЛЬНЫЕ СТАТЬИUnknownThe Membrane Nanodomain Flot1 Protein Participates in Formation of the Early Endosomes in the Root Cells of Arabidopsis thalianaБелок мембранных нанодоменов Flot1 участвует в образовании ранних эндосом в клетках корней A. thalianaKhalilovaL. A.ХалиловаЛ. А.
Russian Federation
lhalilova@mail.ruVoronkovA. S.ВоронковА. С.
Russian Federation
lhalilova@mail.ruTimiryazev Institute of Plant Physiology, Russian Academy of SciencesФедеральное государственное бюджетное учреждение науки Институт физиологии растений им. К.А. Тимирязева Российской академии наук0107202370438239128012025Copyright ©; 2023, Khalilova L.A., Voronkov A.S.Copyright ©; 2023, Л.А. Халилова, А.С. Воронков2023Khalilova L.A., Voronkov A.S.Л.А. Халилова, А.С. Воронковhttps://journals.eco-vector.com/0015-3303/article/view/648147

Plants are subjected to various stress factors within their lifespan. In this respect, the plasma membrane is a principal cell compartment responsible for plant adaptations to stresses. It is capable of remodeling its protein composition by means of endocytosis. In the plants, the main mode of this process is a clathrinmediated endocytosis. Several clathrin-independent pathways are also known; these alternative mechanisms involve Flot1 protein. In the present research, the role of Flot1 in the endocytosis process was examined in seedling roots of a wild type and an Atflot1ko knockout mutant of Arabidopsis thaliana (L.) Heynh. Light microscopy with an FM4-64 lipophilic probe and transmission electron microscopy were used. It was found that endocytosis was arrested in the root cells of the wild type after a simultaneous treatment of the roots with an inhibitor of clathrin-mediated endocytosis (1-naphthylacetic acid) and the agent depleting the plasma membrane of sterols (methyl-β-cyclodextrin). In this case, such morphological change as reduction in cytoplasm vesiculation (including the early endosomes, the small vesicles originated from the agranular ER, the microvacuoles from its fragments, and the clathrin vesicles) was observed. The vesiculation was diminished in both the control and the stressed plants (exposed to 100 mM NaCl). In the Atflot1ko mutant, the cisterns of the Golgi complex closed up to a ring, and the process of formation of the early endosomes was completely abolished under these conditions. It is suggested that, in the roots of A. thaliana exposed to the inhibitors, the microdomain-associated Flot1 protein of the plasma membrane conserves the structure of the Golgi complex and its capacity to build early endosomes on the trans-side. In addition, the protein appears to participate in formation of the early endosomes from the trans-Golgi network.

Растения в ходе всей своей жизни подвергаются различным стрессовым воздействиям. Плазматическая мембрана – основной компартмент клетки, отвечающий за адаптацию растений к стрессу и способный ремоделировать состав входящих в нее белков посредством эндоцитоза. Основным путем эндоцитоза у растений служит клатрин-зависимый эндоцитоз, но кроме него также известны несколько альтернативных, клатрин-независимых путей эндоцитоза. Одним из белков, участвующих в таком эндоцитозе, является Flot1. В данном исследовании на модельной системе корней проростков A. thaliana дикого типа и нокаут-мутанта (Atflot1ko) методами световой микроскопии с флуоресцентным липофильным зондом FM4-64 и электронной микроскопии была исследована роль Flot1 в процессе эндоцитоза. Одновременная обработка корней проростков ингибитором клатринового эндоцитоза (1-нафталинуксусной кислотой) и агентом, обедняющим плазмалемму по стеринам (метил-β-циклодекстрином), блокировала эндоцитоз в клетках корней дикого типа. Вследствие этого, наблюдалось снижение везикуляции цитоплазмы (ранних эндосом, мелких везикул, образованных агранулярным эндоплазматическим ретикулумом, и микровакуолей из его фрагментов, а также клатриновых везикул) как в контрольном варианте, так и в условиях стресса – при внесении в среду 100 мМ NaCl. При этом у мутанта Atflot1ko происходило замыкание цистерн комплекса Гольджи в кольцо с полным блокированием процесса формирования ранних эндосом. Таким образом, полученные результаты наводят на мысль, что при действии ингибиторов на клетки корней A. thaliana присутствующий в микродоменах плазматической мембраны белок Flot1 сохраняет структуру комплекса Гольджи и его способность к формированию на транс-стороне ранних эндосом, а также принимает участие в образовании ранних эндосом из транс-Гольджи сети.

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