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Interactome analysis reveals versatile functions of Arabidopsis COLD SHOCK DOMAIN PROTEIN 3 in RNA processing within the nucleus and cytoplasm

机译:相互作用分析显示拟南芥冷休克域蛋白3在细胞核和细胞质内的RNA加工中具有多种功能

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Arabidopsis COLD SHOCK DOMAIN PROTEIN 3 (AtCSP3) shares an RNA chaperone function with E. coli cold shock proteins and regulates freezing tolerance during cold acclimation. Here, we screened for AtCSP3-interacting proteins using a yeast two-hybrid system and 38 candidate interactors were identified. Sixteen of these were further confirmed in planta interaction between AtCSP3 by a bi-molecular fluorescence complementation assay. We found that AtCSP3 interacts with CONSTANS-LIKE protein 15 and nuclear poly(A)-binding proteins in nuclear speckles. Three 60S ribosomal proteins (RPL26A, RPL40A/UBQ2, and RPL36aB) and the Gar1 RNA-binding protein interacted with AtCSP3 in the nucleolus and nucleoplasm, suggesting that AtCSP3 functions in ribosome biogenesis. Interactions with LOS2/enolase and glycine-rich RNA-binding protein 7 that are cold inducible, and an mRNA decapping protein 5 (DCP5) were observed in the cytoplasm. These data suggest that AtCSP3 participates in multiple complexes that reside in nuclear and cytoplasmic compartments and possibly regulates RNA processing and functioning.
机译:拟南芥冷休克蛋白3(AtCSP3)与大肠杆菌冷休克蛋白具有RNA伴侣功能,并在冷驯化过程中调节冷冻耐受性。在这里,我们使用酵母双杂交系统筛选了与AtCSP3相互作用的蛋白质,并确定了38个候选相互作用子。通过双分子荧光互补测定进一步证实了其中的16种在AtCSP3之间的植物相互作用中。我们发现AtCSP3与CONSTANS-LIKE蛋白15和核斑点中的核poly(A)结合蛋白相互作用。三种60S核糖体蛋白(RPL26A,RPL40A / UBQ2和RPL36aB)和Gar1 RNA结合蛋白在核仁和核质中与AtCSP3相互作用,表明AtCSP3在核糖体生物发生中起作用。在细胞质中观察到与LOS2 /烯醇酶和冷诱导的富含甘氨酸的RNA结合蛋白7的相互作用以及mRNA脱盖蛋白5(DCP5)的相互作用。这些数据表明AtCSP3参与驻留在核和细胞质区室的多种复合物,并可能调节RNA的加工和功能。

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