Kim Myung-Hee, Sasaki Kentaro, Imai Ryozo
Crop Cold Tolerance Research Team, National Agricultural Research Center for Hokkaido Region, Hitsujigaoka 1, Toyohira-ku, Sapporo 062-8555, Japan.
J Biol Chem. 2009 Aug 28;284(35):23454-60. doi: 10.1074/jbc.M109.025791. Epub 2009 Jun 25.
In response to cold, Escherichia coli produces cold shock proteins (CSPs) that have essential roles in cold adaptation as RNA chaperones. Here, we demonstrate that Arabidopsis cold shock domain protein 3 (AtCSP3), which shares a cold shock domain with bacterial CSPs, is involved in the acquisition of freezing tolerance in plants. AtCSP3 complemented a cold-sensitive phenotype of the E. coli CSP quadruple mutant and displayed nucleic acid duplex melting activity, suggesting that AtCSP3 also functions as an RNA chaperone. Promoter-GUS transgenic plants revealed tissue-specific expression of AtCSP3 in shoot and root apical regions. When exposed to low temperature, GUS activity was extensively induced in a broader region of the roots. In transgenic plants expressing an AtCSP3-GFP fusion, GFP signals were detected in both the nucleus and cytoplasm. An AtCSP3 knock-out mutant (atcsp3-2) was sensitive to freezing compared with wild-type plants under non-acclimated and cold-acclimated conditions, whereas expression of C-repeat-binding factors and their downstream genes during cold acclimation was not altered in the atcsp3-2 mutant. Overexpression of AtCSP3 in transgenic plants conferred enhanced freezing tolerance over wild-type plants. Together, the data demonstrated an essential role of RNA chaperones for cold adaptation in higher plants.
为响应寒冷,大肠杆菌会产生冷休克蛋白(CSPs),这些蛋白作为RNA伴侣在冷适应过程中发挥着重要作用。在此,我们证明拟南芥冷休克结构域蛋白3(AtCSP3)与细菌CSPs共享一个冷休克结构域,它参与了植物抗冻性的获得。AtCSP3弥补了大肠杆菌CSP四突变体的冷敏感表型,并表现出核酸双链解链活性,这表明AtCSP3也作为RNA伴侣发挥作用。启动子-GUS转基因植物显示AtCSP3在茎尖和根尖区域有组织特异性表达。当暴露于低温时,根的更广泛区域中GUS活性被广泛诱导。在表达AtCSP3-GFP融合蛋白的转基因植物中,在细胞核和细胞质中均检测到GFP信号。与野生型植物相比,在非驯化和冷驯化条件下,AtCSP3敲除突变体(atcsp3-2)对冷冻敏感,而在atcsp3-2突变体中,冷驯化过程中C-重复结合因子及其下游基因的表达未发生改变。转基因植物中AtCSP3的过表达赋予了比野生型植物更强的抗冻性。总之,这些数据证明了RNA伴侣在高等植物冷适应中的重要作用。