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甘氨酸丰富的 RNA 结合蛋白基因在荠蓝中的分子克隆、鉴定及应激表达分析

Molecular cloning, characterization, and stress-responsive expression of genes encoding glycine-rich RNA-binding proteins in Camelina sativa L.

机构信息

Bioenergy Research Center, Department of Bioenergy Science and Technology, Chonnam National University, Gwangju 500-757, South Korea.

出版信息

Plant Physiol Biochem. 2013 Jul;68:44-51. doi: 10.1016/j.plaphy.2013.03.023. Epub 2013 Apr 11.

Abstract

Camelina sativa L. is an oil-seed crop that has potential for biofuel applications. Although the importance of C. sativa as a biofuel crop has increased in recent years, reports demonstrating the stress responsiveness of C. sativa and characterizing the genes involved in stress response of C. sativa have never been published. Here, we isolated and characterized three genes encoding glycine-rich RNA-binding proteins (GRPs) from camelina: CsGRP2a, CsGRP2b, and CsGRP2c. The three CsGRP2 proteins were very similar in amino acid sequence and contained a well-conserved RNA-recognition motif at the N-terminal region and glycine-rich domain at the C-terminal region. To understand the functional roles of CsGRP2s under stress conditions, we investigated the expression patterns of CsGRP2s under various environmental stress conditions. The expressions of the three CsGRP2s were highly up-regulated under cold stress. The expression of CsGRP2a was up-regulated under salt or dehydration stress, whereas the transcript levels of CsGRP2b and CsGRP2c were decreased under salt or dehydration stress conditions. The three CsGRP2s had the ability to complement cold-sensitive Escherichia coli mutants at low temperatures and harbored transcription anti-termination and nucleic acid-melting activities, indicating that the CsGRP2s possess RNA chaperone activity. The CsGRP2a protein was localized to both the nucleus and the cytoplasm. Expression of CsGRP2a in cold-sensitive Arabidopsis grp7 mutant plants resulted in decreased electrolyte leakage at freezing temperatures. Collectively, these results suggest that the stress-responsive CsGRP2s play a role as an RNA chaperone during the stress adaptation process in camelina.

摘要

荠蓝是一种潜在的生物燃料用含油种子作物。尽管荠蓝作为生物燃料作物的重要性近年来有所增加,但尚未有报道表明荠蓝对压力的反应,以及描述荠蓝压力反应中涉及的基因。在这里,我们从荠蓝中分离并鉴定了三个编码甘氨酸丰富的 RNA 结合蛋白(GRPs)的基因:CsGRP2a、CsGRP2b 和 CsGRP2c。这三个 CsGRP2 蛋白在氨基酸序列上非常相似,在 N 端区域含有保守的 RNA 识别基序,在 C 端区域含有甘氨酸丰富的结构域。为了了解 CsGRP2 在胁迫条件下的功能作用,我们研究了 CsGRP2 在各种环境胁迫条件下的表达模式。在冷胁迫下,这三个 CsGRP2 的表达均高度上调。CsGRP2a 的表达在盐或脱水胁迫下上调,而 CsGRP2b 和 CsGRP2c 的转录水平在盐或脱水胁迫条件下降低。这三个 CsGRP2 能够在低温下互补冷敏感的大肠杆菌突变体,并具有转录抗终止和核酸解链活性,表明 CsGRP2 具有 RNA 伴侣活性。CsGRP2a 蛋白定位于细胞核和细胞质。在冷敏感的拟南芥 grp7 突变体植物中表达 CsGRP2a 导致在冷冻温度下电解质泄漏减少。总之,这些结果表明,胁迫响应型 CsGRP2s 在荠蓝适应胁迫的过程中作为 RNA 伴侣发挥作用。

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