Differential regulation of Ku gene expression in etiolated mung bean hypocotyls by auxins

Pei-Feng Liu , Wen-Chi Chang , Yung-Kai Wang , Suresh-Babu Munisamy , Shen-Hsing Hsu , Hwan-You Chang , Shu-Hsing Wu , Rong-Long Pan
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引用次数: 10

Abstract

Plant Ku genes were identified very recently in Arabidopsis thaliana, and their roles in repair of double-stranded break DNA and maintenance of telomere integrity were scrutinized. In this study, the cDNAs encoding Ku70 (VrKu70) and Ku80 (VrKu80) were isolated from mung bean (Vigna radiata L.) hypocotyls. Both genes were expressed widely among different tissues of mung bean with the highest levels in hypocotyls and leaves. The VrKu gene expression was stimulated by exogenous auxins in a concentration- and time-dependent manner. The stimulation could be abolished by auxin transport inhibitors, N-(1-naphthyl) phthalamic acid and 2,3,5-triiodobenzoic acid implicating that exogenous auxins triggered the effects following their uptake by the cells. Further analysis using specific inhibitors of auxin signaling showed that the stimulation of VrKu expression by 2,4-dichlorophenoxyacetic acid (2,4-D) was suppressed by intracellular Ca2+ chelators, calmodulin antagonists, and calcium/calmodulin dependent protein kinase inhibitors, suggesting the involvement of calmodulin in the signaling pathway. On the other hand, exogenous indole-3-acetic acid (IAA) and α-naphthalene acetic acid (NAA) stimulated VrKu expression through the mitogen-activated protein kinase/extracellular signal-regulated kinase pathway. Altogether, it is thus proposed that 2,4-D and IAA (or NAA) regulate the expression of VrKu through two distinct pathways.

生长素对黄化绿豆下胚轴Ku基因表达的差异调控
植物Ku基因最近在拟南芥中被发现,并在双链断裂DNA修复和端粒完整性维持中发挥了重要作用。本研究从绿豆(Vigna radiata L.)下胚轴中分离到了编码Ku70 (VrKu70)和Ku80 (VrKu80)的cdna。这两个基因在绿豆的不同组织中广泛表达,在下胚轴和叶片中表达量最高。外源生长素刺激VrKu基因表达呈浓度依赖性和时间依赖性。这种刺激可以被生长素运输抑制剂N-(1-萘基)邻苯二甲酸和2,3,5-三碘苯甲酸所消除,这表明外源生长素在被细胞摄取后触发了这种作用。利用生长素信号传导的特异性抑制剂进一步分析表明,2,4-二氯苯氧乙酸(2,4- d)对VrKu表达的刺激被细胞内Ca2+螯合剂、钙调素拮抗剂和钙/钙调素依赖性蛋白激酶抑制剂抑制,表明钙调素参与信号通路。另一方面,外源吲哚-3-乙酸(IAA)和α-萘乙酸(NAA)通过丝裂原活化蛋白激酶/细胞外信号调节激酶途径刺激VrKu表达。综上所述,2,4- d和IAA(或NAA)通过两种不同的途径调控VrKu的表达。
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