Interactome analysis reveals versatile functions of Arabidopsis COLD SHOCK DOMAIN PROTEIN 3 in RNA processing within the nucleus and cytoplasm
Interactome analysis reveals versatile functions of Arabidopsis COLD SHOCK DOMAIN PROTEIN 3 in RNA processing within the nucleus and cytoplasm
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DOI：
10.1007/s12192-012-0398-3
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发表时间：
2013-07-01
影响因子：
3.8
通讯作者：
Imai, Ryozo
中科院分区：
文献类型：
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作者：
Kim, Myung-Hee;Sonoda, Yutaka;Imai, Ryozo
拟南芥冷休克结构域蛋白3(AtCSP3)与大肠杆菌冷休克蛋白具有共同的RNA伴侣功能，调节冷驯化过程中的耐寒性。在这里，我们使用酵母双杂交系统筛选AtCSP3相互作用蛋白，并鉴定出38个候选相互作用蛋白。通过双分子荧光互补分析进一步证实了其中16个基因在AtCSP3之间的相互作用。我们发现AtCSP3与核斑点中的Constans-like Protein 15和核Poly(A)结合蛋白相互作用。3个60S核糖体蛋白(RPL26A、RPL40A/UBQ2和RPL36aB)以及Gar1 RNA结合蛋白与AtCSP3在核仁和核质中相互作用，表明AtCSP3在核糖体生物发生中发挥作用。在细胞质中观察到与冷诱导的LOS2/烯醇化酶和富含甘氨酸的RNA结合蛋白7以及mRNA解帽蛋白5(DCP5)的相互作用。这些数据表明，AtCSP3参与了位于核间和细胞质间的多个复合体，并可能调节RNA的加工和功能。
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.