Zhang Yuxiu, Li Jinmei, Yu Fei, Cong Lin, Wang Liyan, Burkard Gérard, Chai Tuanyao
Department of Biological Engineering, School of Chemical and Environmental Engineering, China University of Mining and Technology (Beijing), Xueyuan Road 11, Beijing 100083, China.
Mol Biotechnol. 2006 Mar;32(3):205-18. doi: 10.1385/MB:32:3:205.
A heavy metal responsive gene PvSR3 (GenBank accession number U54703) encoding an acid dehydrin was isolated from a mercuric chloride-treated bean (Phaseolus vulgaris L.) leaf cDNA library by differential screening using cDNAs derived from treated and untreated plants. The PvSR3 cDNA is 981-bp long and has a 606-bp open-reading frame with a 202-residue-deduced amino acid sequence. The PvSR3 sequence contains two conserved repeats of the characteristic lysine-rich K segment (EKKGIMDKIKEKLPG) preceded by an 8-serine residue stretch, whereas the Y segment (DEYGNP) conserved motif is absent. The deduced protein has a calculated molecular weight of 23 kDa and an isoelectric point of 5.2. Sequence similarity and comparative analysis showed that PvSR3 shares 70 and 73% similarity with the dehydrin of poplar and pepper, respectively. Southern hybridizations indicated that PvSR3 was a low copy-number gene. Northern blot analysis revealed that PvSR3 mRNA was weakly detected in seedling leaves. However, the gene expression was strongly stimulated by heavy metals, such as mercury, cadmium, arsenic, and copper, whereas virus infection and salt had little effect on it. In contrast, PvSR3 was not responsive to drought or abscisic acid (ABA), and was downregulated by UV radiation. Furthermore, PvSR3 was upregulated by the exogenous signaling molecules, including salicylic acid (SA) and hydrogen peroxide (H2O2). It is suggested that PvSR3 is extremely related to heavy metal stress, and might play an important role in metal detoxification and resistance to the damage caused by heavy metals.
从经氯化汞处理的菜豆(Phaseolus vulgaris L.)叶片cDNA文库中,通过差异筛选法,利用来自处理和未处理植株的cDNA,分离出了一个编码酸性脱水素的重金属响应基因PvSR3(GenBank登录号U54703)。PvSR3 cDNA长981 bp,有一个606 bp的开放阅读框,推导的氨基酸序列有202个残基。PvSR3序列包含两个富含赖氨酸的特征性K片段(EKKGIMDKIKEKLPG)的保守重复序列,其前面有一段8个丝氨酸残基的延伸,而Y片段(DEYGNP)保守基序缺失。推导的蛋白质计算分子量为23 kDa,等电点为5.2。序列相似性和比较分析表明,PvSR3与杨树和辣椒的脱水素分别有70%和73%的相似性。Southern杂交表明PvSR3是一个低拷贝数基因。Northern印迹分析显示,在幼苗叶片中微弱检测到PvSR3 mRNA。然而,该基因的表达受到汞、镉、砷和铜等重金属的强烈刺激,而病毒感染和盐对其影响很小。相反,PvSR3对干旱或脱落酸(ABA)无响应,且受紫外线辐射下调。此外,PvSR3受外源信号分子上调,包括水杨酸(SA)和过氧化氢(H2O2)。提示PvSR3与重金属胁迫密切相关,可能在金属解毒和抵抗重金属造成的损伤中起重要作用。